U.S. patent application number 16/863187 was filed with the patent office on 2020-08-13 for equol-containing extract, method for production thereof, method for extraction of equol, and equol-containing food.
This patent application is currently assigned to OTSUKA PHARMACEUTICAL CO., LTD.. The applicant listed for this patent is OTSUKA PHARMACEUTICAL CO., LTD.. Invention is credited to Izumi HANYA, Takuma IMADA, Kyoko KAMEDA, Takuya KISHIMOTO, Toshiaki MATSUMOTO, Kunihiko MATSUOKA, Kentaro TADANO, Takao TAKI, Michiaki TOMINAGA, Machiko TSUJI, Hiromasa TSUNEISHI, Yuko UCHIYAMA, Tania VALDES-GONZALEZ.
Application Number | 20200253251 16/863187 |
Document ID | 20200253251 / US20200253251 |
Family ID | 1000004794694 |
Filed Date | 2020-08-13 |
Patent Application | download [pdf] |
United States Patent
Application |
20200253251 |
Kind Code |
A1 |
TOMINAGA; Michiaki ; et
al. |
August 13, 2020 |
EQUOL-CONTAINING EXTRACT, METHOD FOR PRODUCTION THEREOF, METHOD FOR
EXTRACTION OF EQUOL, AND EQUOL-CONTAINING FOOD
Abstract
The present invention relates to an extract obtained by
extracting useful components containing equol from an
equol-containing fermented soybean hypocotyl, and to a method for
producing the same. The present invention makes it possible to
efficiently obtain useful components containing equol from the
fermented soybean hypocotyl by subjecting an equol-containing
fermented soybean hypocotyl to extraction using an ethanol aqueous
solution as an extractant. The present invention reduces the
content of saponin, which causes an unpleasant taste, by
sequentially subjecting the equol-containing fermented soybean
hypocotyl to extraction using an ethanol aqueous solution and
ethanol, while efficiently extracting equol and glycitein.
Inventors: |
TOMINAGA; Michiaki; (Osaka,
JP) ; TAKI; Takao; (Osaka, JP) ; MATSUMOTO;
Toshiaki; (Osaka, JP) ; HANYA; Izumi; (Osaka,
JP) ; IMADA; Takuma; (Osaka, JP) ; MATSUOKA;
Kunihiko; (Osaka, JP) ; KISHIMOTO; Takuya;
(Osaka, JP) ; UCHIYAMA; Yuko; (Osaka, JP) ;
TSUNEISHI; Hiromasa; (Otsu-shi, JP) ; TSUJI;
Machiko; (Otsu-shi, JP) ; VALDES-GONZALEZ; Tania;
(Osaka, JP) ; TADANO; Kentaro; (Osaka, JP)
; KAMEDA; Kyoko; (Osaka, JP) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
OTSUKA PHARMACEUTICAL CO., LTD. |
Tokyo |
|
JP |
|
|
Assignee: |
OTSUKA PHARMACEUTICAL CO.,
LTD.
Tokyo
JP
|
Family ID: |
1000004794694 |
Appl. No.: |
16/863187 |
Filed: |
April 30, 2020 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
13673617 |
Nov 9, 2012 |
10681930 |
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16863187 |
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12664298 |
Dec 11, 2009 |
8900645 |
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PCT/JP2008/060913 |
Jun 13, 2008 |
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13673617 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A23L 2/52 20130101; A23C
9/1544 20130101; A23C 11/103 20130101; A23F 3/163 20130101; A23C
11/02 20130101; A61K 36/48 20130101; A23L 23/00 20160801; A23L
27/60 20160801; A21D 2/368 20130101; A23L 11/09 20160801; A23L 5/23
20160801; A23L 21/15 20160801; A23C 9/1315 20130101; A61K 31/353
20130101; A23L 33/105 20160801; A23L 13/65 20160801; A23C 20/025
20130101; A23L 17/70 20160801; A23F 5/243 20130101; A23C 9/1307
20130101; A23L 9/12 20160801; A23L 7/196 20160801; A23L 27/50
20160801; A23L 13/426 20160801 |
International
Class: |
A23L 11/00 20060101
A23L011/00; A21D 2/36 20060101 A21D002/36; A23C 9/13 20060101
A23C009/13; A23C 9/154 20060101 A23C009/154; A23C 11/02 20060101
A23C011/02; A23C 11/10 20060101 A23C011/10; A23C 20/02 20060101
A23C020/02; A23F 3/16 20060101 A23F003/16; A23F 5/24 20060101
A23F005/24; A23L 2/52 20060101 A23L002/52; A61K 31/353 20060101
A61K031/353; A61K 36/48 20060101 A61K036/48; A23L 9/10 20060101
A23L009/10; A23L 5/20 20060101 A23L005/20; A23L 17/00 20060101
A23L017/00; A23L 33/105 20060101 A23L033/105; A23L 7/196 20060101
A23L007/196; A23L 13/40 20060101 A23L013/40; A23L 21/15 20060101
A23L021/15; A23L 27/60 20060101 A23L027/60; A23L 27/50 20060101
A23L027/50; A23L 23/00 20060101 A23L023/00; A23L 13/60 20060101
A23L013/60 |
Foreign Application Data
Date |
Code |
Application Number |
Jun 13, 2007 |
JP |
2007-156822 |
Jun 13, 2007 |
JP |
2007-156825 |
Jun 13, 2007 |
JP |
2007-156833 |
Claims
1-26. (canceled)
27. A drink composition comprising (a) and (b): (a) equol; and (b)
ornithine, wherein the equol and ornithine are present in a
fermented substance obtained by fermenting a material containing at
least one daidzein compound selected from the group consisting of
daidzein glycosides, daidzein and dihydrodaidzein.
28. The drink composition according to claim 27, wherein the drink
composition is at least one member selected from the group
consisting of sparkling water, orange-flavored carbonated beverage
containing no orange juice, low-calorie cola, carbonated apple
beverage, carbonated grape beverage, beverage containing orange
juice, low calorie beverage containing fruit juice, sugarless near
water, sports drink, yogurt drink, lactic acid bacteria beverage,
low-calorie sour milk beverage, cream soda, beverage containing
apple juice, soft drink, beverage containing orange juice, oolong
tea, tea with lemon, sour milk beverage, beverage containing
vitamins, green tea, powered orange juice drink, amino acid drink
and energy drink.
29. The drink composition according to claim 27, wherein the drink
composition contains (c) at least one member selected from the
group consisting of citric acid, sodium citrate, ascorbic acid,
sodium L-ascorbate, lactic acid and malic acid, in a range of 0.003
to 5 wt %.
30. A yoghurt composition comprising (a), (b) and (d): (a) equol;
(b) ornithine; and, (d) starter yoghurt, wherein the equol and
ornithine are present in a fermented substance obtained by
fermenting a material containing at least one daidzein compound
selected from the group consisting of daidzein glycosides, daidzein
and dihydrodaidzein.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This is a Divisional of U.S. patent application Ser. No.
12/664,298 filed Dec. 11, 2009, which is a 371 National Stage Entry
of PCT/JP2008/060913 filed Jun. 13, 2008, which claims priority of
Japanese Patent Application Nos. 2007-156822, 2007-156825, and
2007-156833 all filed Jun. 13, 2007. The entire contents of the
aforementioned applications are incorporated herein by
reference.
TECHNICAL FIELD
[0002] The present invention relates to an equol-containing extract
obtained by extracting effective components from an
equol-containing fermented soybean hypocotyl and a production
method thereof. The present invention also relates to a method for
efficiently purifying an equol-containing substance to obtain a
highly pure equol. The present invention further relates to an
equol-containing food material and an equol-containing food.
BACKGROUND ART
[0003] The isoflavones (soybean isoflavones: daidzein, genistein,
glycitein) contained in soybeans have structures similar to
estradiol, and have anti-estrogen actions associated with binding
to estrogen receptors (hereinafter referred to as ER) and
estrogen-like actions. The ever conducted epidemiological studies
and intervention studies of soybean isoflavones suggest that they
have preventive effects due to their anti-estrogen actions on
breast cancer, prostate cancer and other hormone-dependent cancers
and improving effects due to their estrogen-like actions on
menopausal disorders, postmenopausal osteoporosis and
hyperlipidemia.
[0004] Recently, it has been pointed out that the active component
of the physiological effects of these soybean isoflavones may be a
metabolite of daidzein, i.e., equol. More specifically, it has been
reported that equol has an ability to bind to ER (especially to
ER.beta.) greater than soybean isoflavones and that it has a
remarkably high transition capability to target organs such as
breast and prostate tissues (Non-Patent Documents 1 to 4).
Moreover, a case-control study reports that there are significantly
less patients who produce equol in the patients of breast cancer
and prostate cancer. The effects of soybean isoflavones that
improve bone density and lipid metabolism were examined regarding
postmenopausal women categorized into two groups: those who produce
equol and those who do not. A significant improvement in those who
produce equol was observed.
[0005] Equol is produced by metabolism of daidzein by enteric
bacteria. The abilities to produce equol vary between individuals,
and the percentage of Japanese who produce equol is reportedly
about 50%. That is, about 50% of Japanese are not able to produce
equol (non-equol-producing individuals). Such an individual cannot
enjoy any useful physiological benefits based on the action of
equol even if they ingest soybeans and processed soybean foods.
Therefore, in order to attain useful physiological benefits based
on the action of equol in a non-equol-producing individual,
ingesting equol itself is thought to be effective. [0006]
Non-patent Document 1: Morito K, Hirose T, Kinjo J, Hirakawa T,
Okawa M, Nohara T, Ogawa S, Inoue S, Muramatsu M, Masamune Y.
Interaction of phytoestrogens with estrogen receptors .alpha. and
.beta. Biol Pharm Bull 24(4): 351-356, 2001 [0007] Non-patent
Document 2: Maubach J, Bracke M E, Heyerick A, Depypere H T,
Serreyn R F, Mareel M M, Keukeleire D D. Quantitation of
soy-derived phytoestrogens in human breast tissue and biological
fluids by high-performance liquid chromatography. J Chromatography
B 784: 137-144, 2003 [0008] Non-patent Document 3: Morton M S, Chan
P S F, Cheng C, Blacklock N, Matos-Ferreira A, Abranches-Monteiro
L, Correia R, Lloyd S, Griffiths K. Lignans and isoflavonoids in
plasma and prostatic fluid in men: Samples from Portugal, Hong
Kong, and the United Kingdom. Prostate 32: 122-128, 1997 [0009]
Non-patent Document 4: Tammy E H, Paul D M, Paul G F, Robert D,
Stephen B, Kenneth J, Ray M, Lorraine G O, Kristiina W, Holly M S,
Karen J G. Long-term dietary habits affect soy isoflavone
metabolism and accumulation in prostatic fluid in caucasian men. J
Nutr 135: 1400-1406, 2005.
DISCLOSURE OF THE INVENTION
Problems to be Solved by the Invention
[0010] The present inventors have found that fermented soybean
hypocotyl obtained by fermenting soybean hypocotyl with an
equol-producing microorganism is usable as an equol-containing food
material. The fermented soybean hypocotyl contains not only equol
but also isoflavone, saponin and like useful soybean-derived
components. The fermented soybean hypocotyl achieves such effective
physiological effects attributable to these useful components, and
therefore is usable as a functional material. The present inventors
also found that the fermented soybean hypocotyl is useful as an
allergen-reduced material, because the allergens attributable to
soybean hypocotyl therein are reduced. The fermented soybean
hypocotyl found by the present inventors exhibits physiological
effects attributable to the useful components, and is
hypoallergenic, and therefore it is usable as a functional food
material.
[0011] In contrast, the content of the equol in the fermented
soybean hypocotyl varies depending on the types of the soybean
hypocotyl used for the production of the fermented soybean
hypocotyl, types of the equol-producing microorganism, etc., but it
is generally about 1 wt. %. If a material with an increased equol
content can be provided, various types of equol-containing foods
become easily available in response to the diversification of
foods. However, the fermented soybean hypocotyl itself is not a
known product, and the effective method for efficiently extracting
useful component containing equol from fermented soybean hypocotyl
has not been found yet.
[0012] The fermented soybean hypocotyl, which is an
equol-containing substance obtained by the fermentation method
describe above, is safer than that obtained by a chemical synthesis
method, and suitable for industrial production. However,
equol-containing substances obtained by said fermentation method
contain metabolites other than equol and various components derived
from raw materials remaining therein. Depending on the type of raw
material used for fermentation, the equol-containing substance
obtained by a fermentation method may contain material that could
become an allergen. Therefore, in order to use an equol as an
additive for foods or drugs, a technique not only for producing an
equol but also purifying the equol-containing substance so as to
obtain a highly pure equol is required. However, not many
techniques for refining equol have been reported yet, there is a
demand for establishing a technique, which is industrially
applicable and by which an equol can be purified easily and
efficiently.
[0013] The fermented soybean hypocotyl has bitterness attributable
to soybean hypocotyl or equol. Therefore, sufficient attention must
be paid to the flavor of the resulting food when the fermented
soybean hypocotyl is used as a food material. The fermented soybean
hypocotyl is a novel food material that has not been reported
before, and it is unknown which method is effective for improving
the flavors thereof. In particular, when the fermented soybean
hypocotyl is used as a material for baked confectionaries, it tends
to suffer from a deterioration of flavor after being subjected to a
baking step. Therefore, exhibiting good flavor is a very difficult
technical object.
[0014] An object of the present invention is to provide an extract,
which contains useful equol-containing components and which is
derived from an equol-containing fermented soybean hypocotyl, and a
method for producing such an extract. The present invention
provides a method for purifying an equol-containing substance to
effectively obtain a highly pure equol. Another object of the
present invention is to provide a food material obtained by
fermenting an equol-containing fermented soybean hypocotyl with an
equol-producing microorganism or its extract, in which the flavor
is improved. Still another object of the present invention is to
provide a food product that contains an equol-containing fermented
soybean hypocotyl and exhibits excellent flavor (in particular,
baked confectionaries). Still another object of the present
invention is to provide various forms of foods that contain an
equol-containing fermented soybean hypocotyl or its extract.
Means for Solving the Problem
[0015] The present inventors conducted intensive research to
achieve the above objects and found that by subjecting an
equol-containing fermented soybean hypocotyl to extraction using an
ethanol aqueous solution as an extractant, a useful
equol-containing component can be efficiently extracted from the
fermented soybean hypocotyl. The present inventors further found
that by sequentially subjecting the equol-containing fermented
soybean hypocotyl to extraction using an ethanol aqueous solution
and ethanol, the saponin content that causes an astringent taste,
bitterness, an acrid taste and like unpleasant tastes can be
reduced, while effectively extracting equol and glycitein.
[0016] The present inventors also found that a highly pure equol
can be obtained by suspending an equol-containing fermented soybean
hypocotyl in an aqueous solution having a pH of 2 to 6, collecting
insoluble matter, and then subjecting the resulting insoluble
residue to extraction using an ethanol aqueous solution.
[0017] The present inventors further found that a highly pure equol
can be efficiently and readily obtained by conducting the following
Steps (1) to (5): [0018] Step (1): subjecting an equol-containing
substance to extraction using an ethanol aqueous solution; [0019]
Step (2): subjecting the resulting extract to further extraction
using ethanol; [0020] Step (3): subjecting the resulting extract to
extraction using hexane; [0021] Step (4): subjecting the resulting
insoluble residue to extraction using a mixture of hexane and
ether; and [0022] Step (5): subjecting the resulting liquid extract
to silica gel column chromatography, and obtaining an
equol-containing fraction.
[0023] The present inventors further found that a highly pure equol
can be efficiently and readily obtained by conducting the following
Steps (i) to (iii): [0024] (i) subjecting the equol-containing
substance to extraction using a specific organic solvent or an
aqueous organic solvent, and concentrating the resulting liquid
extract; [0025] (ii) subjecting the resulting equol concentrate to
silica gel column chromatography, and obtaining an equol-containing
fraction; and [0026] (iii) removing the solvent from the resulting
equol-containing fraction, and recrystallizing the resulting
residue.
[0027] The present inventors also found that in a food material
obtained by dispersing an equol-containing fermented soybean
hypocotyl or its extract in cacao mass, the bitterness attributable
to the equol-containing fermented soybean hypocotyl can be reduced
and excellent flavor can be exhibited. The present inventors also
found that the food material is usable in various foods, including
baked confectionaries, without adversely affecting the flavor.
[0028] The present inventors successfully developed various food
products into which an equol-containing fermented soybean hypocotyl
or its extract is added.
[0029] The present invention has been accomplished based on these
findings and some improvement added thereto.
[0030] The present invention provides a method for obtaining an
equol-containing extract.
[0031] Item 1-1. A method for producing an equol-containing extract
comprising Step I-1: [0032] subjecting an equol-containing
fermented soybean hypocotyl to extraction using an ethanol aqueous
solution as an extractant; and collecting the resulting liquid
extract.
[0033] Item 1-2. The method according to Item 1-1, which further
comprises Step I-2: [0034] removing the extractant from the liquid
extract obtained in Step I-1; [0035] conducting extraction using
ethanol as an extractant; and collecting the liquid extract.
[0036] Item 1-3. The method according to Item 1-1, wherein the
ethanol aqueous solution used in Step I-1 has an ethanol
concentration of 20 to 98 wt. %.
[0037] Item 1-4. The method according to any one of Items 1-1 to
1-3, wherein the equol-containing fermented soybean hypocotyl is
obtained by fermenting a soybean hypocotyl with an equol-producing
microorganism.
[0038] Item 1-5. A method for producing an equol-containing extract
from equol-containing fermented soybean hypocotyl, comprising Steps
II-1 and III-2; [0039] Step II-1 of suspending the equol-containing
fermented soybean hypocotyl in water, adding an acid so as to
adjust the pH to 2 to 6, and collecting insoluble residue; and
[0040] Step II-2 of subjecting the insoluble residue obtained in
Step II-1 to extraction using an ethanol aqueous solution as an
extractant, and then collecting the resulting liquid extract.
[0041] Item 1-6. The method of Item 1-5, wherein the acid used in
Step II-1 is acetic acid.
[0042] The present invention provides the following
equol-containing extracts.
[0043] Item 2-1. An equol-containing extract obtained by subjecting
an equol-containing fermented soybean hypocotyl to extraction using
an ethanol aqueous solution as an extractant.
[0044] Item 2-2. An equol-containing extract obtained by
sequentially subjecting an equol-containing fermented soybean
hypocotyl to extraction using an ethanol aqueous solution and
ethanol.
[0045] Item 2-3. An equol-containing extract obtained by a method
according to any one of Items 1-1 to 1-6.
[0046] The present invention provides the following methods for
refining an equol.
[0047] Item 3-1. A method for purifying an equol-containing
substance to obtain equol comprising Steps 1-1 to 1-5 below: [0048]
Step 1-1 of subjecting an equol-containing substance to extraction
using an ethanol aqueous solution as an extractant, and collecting
the resulting liquid extract; [0049] Step 1-2 of removing the
solvent from the liquid extract obtained in Step 1-1, subjecting
the resulting residue to extraction using ethanol as an extractant,
and then collecting the liquid extract; [0050] Step 1-3 of removing
the solvent from the liquid extract obtained in Step 1-2,
suspending the resulting residue in hexane, and then collecting an
insoluble residue; [0051] Step 1-4 of subjecting the insoluble
residue obtained in Step 1-3 to extraction using a mixture of
hexane and ether as an extractant, and then collecting the liquid
extract; and [0052] Step 1-5 of subjecting the liquid extract
obtained in Step 1-4 to silica gel column chromatography, and
obtaining an equol-containing fraction.
[0053] Item 3-2. The method according to Item 3-1, wherein the
equol-containing substance is a fermented material that contains an
equol.
[0054] Item 3-3. The method according to Item 3-2, wherein the
fermented material that contains an equol is an equol-containing
fermented soybean hypocotyl material obtained by fermenting a
soybean hypocotyl with an equol-producing microorganism.
[0055] Item 3-4. The method according to any one of Items 3-1 to
3-3, wherein the extractant used in Step 1-4 is a mixture of hexane
and diethyl ether.
[0056] Item 3-5. The method according to any one of Items 3-1 to
3-4, wherein the extractant used in Step 1-4 is a mixture of hexane
and ether having a volume ratio of 10:90 to 20:80.
[0057] Item 3-6. The method according to any one of Items 3-1 to
3-5, wherein the ethanol concentration of the ethanol aqueous
solution used in Step 1-1 is 20 to 98 vol. %.
[0058] Item 3-7. A method for purifying an equol-containing
substance comprising Steps 2-1 to 2-3 of: [0059] Step 2-1 of
subjecting the equol-containing substance to extraction using, as
an extractant, an organic solvent or an aqueous organic solvent of
at least one member selected from the group consisting of ethyl
acetate, alcohol, acetone, dioxane, acetonitrile, diethyl ether,
and toluene, and collecting the resulting liquid extract; [0060]
Step 2-2 of subjecting the liquid extract obtained in Step 2-1 to
silica gel column chromatography, and obtaining an equol-containing
fraction; and [0061] Step 2-3 of removing the solvent from the
equol-containing fraction obtained in Step 2-2, recrystallizing the
resulting residue, and collecting the deposited crystals.
[0062] Item 3-8. The method according to Item 3-7, wherein Step 2-3
comprises a recrystallization using an aqueous alcohol solution
after the recrystallization using a mixture of ethyl acetate and
hexane.
[0063] The present invention provides the following food materials,
and foods using such food materials.
[0064] Item 4-1. A food material obtained by dispersing an
equol-containing fermented soybean hypocotyl or its extract in
cacao mass.
[0065] Item 4-2. The food material according to Item 4-1, wherein
the food material contains 10 to 2000 parts by weight of cacao mass
per 100 parts by weight of equol-containing fermented soybean
hypocotyl or its extract.
[0066] Item 4-3. The food material according to Item 4-1 or 4-2,
wherein the equol-containing fermented soybean hypocotyl is
obtained by fermenting soybean hypocotyl with an equol-producing
microorganism.
[0067] Item 4-4. The food material according to Item 4-3, wherein
the equol-producing microorganism is a microorganism that can
metabolize at least one daidzein compound selected from the group
consisting of daidzein glycoside, daidzein, and dihydrodaidzein to
produce equol.
[0068] Item 4-5. The food material according to any one of Items
4-1 to 4-4, which takes a granular form, a chip form, or a
plate-like form.
[0069] Item 4-6. An equol-containing food comprising a food
material of any one of Items 4-1 to 4-5.
[0070] Item 4-7. The equol-containing food according to Item 4-6,
which is a baked confectionary.
[0071] The present invention provides the following foods: Item
5-1. A food comprising an equol-containing fermented soybean
hypocotyl or its extract.
[0072] Item 5-2. A beverage comprising an equol-containing
fermented soybean hypocotyl or its extract.
[0073] Item 5-3. A dietary supplement comprising an
equol-containing fermented soybean hypocotyl or its extract.
[0074] Item 5-4. A creamy food comprising an equol-containing
fermented soybean hypocotyl or its extract.
[0075] Item 5-5. A dessert comprising an equol-containing fermented
soybean hypocotyl or its extract.
[0076] Item 5-6. A snack comprising an equol-containing fermented
soybean hypocotyl or its extract.
[0077] Item 5-7. A seasoning comprising an equol-containing
fermented soybean hypocotyl or its extract.
[0078] Item 5-8. A retort-packed food comprising an
equol-containing fermented soybean hypocotyl or its extract.
[0079] Item 5-9. A processed meat product comprising an
equol-containing fermented soybean hypocotyl or its extract.
[0080] Item 5-10. A fish paste comprising an equol-containing
fermented soybean hypocotyl or its extract.
[0081] Item 5-11. A processed egg product comprising an
equol-containing fermented soybean hypocotyl or its extract.
[0082] Item 5-12. A canned food comprising an equol-containing
fermented soybean hypocotyl or its extract.
[0083] Item 5-13. A bread product comprising an equol-containing
fermented soybean hypocotyl or its extract.
[0084] Item 5-14. A frozen dessert comprising an equol-containing
fermented soybean hypocotyl or its extract.
[0085] Item 5-15. A processed soybean food comprising an
equol-containing fermented soybean hypocotyl or its extract.
[0086] Item 5-16. A cooked rice product comprising an
equol-containing fermented soybean hypocotyl or its extract.
[0087] Item 5-17. A soup comprising an equol-containing fermented
soybean hypocotyl or its extract.
[0088] Item 5-18. A food according to any one of Items 5-1 to 5-18,
wherein the equol-containing fermented soybean hypocotyl is
obtained by fermenting a soybean hypocotyl with an equol-producing
microorganism.
[0089] Item 5-19. The food according to Item 5-18, wherein the
microorganism has an ability to metabolize at least one daidzein
compound selected from the group consisting of daidzein glycoside,
daidzein, and dihydrodaidzein to produce equol.
Effects of the Invention
[0090] The method for producing an equol-containing extract of the
present invention can produce an equol-containing extract that is
useful as a functional food material by effectively extracting
useful components containing equol from the equol-containing
fermented soybean hypocotyl. By sequentially subjecting an
equol-containing fermented soybean hypocotyl to extraction using an
ethanol aqueous solution and ethanol, an equol-containing extract
that contains a high concentration of equol and glycitein, with a
reduced amount of saponin that causes an unpleasant taste is
obtainable. Accordingly, the equol-containing extract can be added
to foods without adversely affecting the taste.
[0091] The purification method of the present invention makes it
possible to readily and efficiently obtain a highly pure equol from
the equol-containing substance. In particular, the purification
method of the present invention makes it possible to highly pure
equol even when the equol-containing substance contains isoflavones
whose structure is similar to that of the equol by removing these
isoflavones. Accordingly, the purification method of the present
invention can be suitably employed in obtaining equol from an
equol-containing fermented material that contains a large
proportion of isoflavones.
[0092] Furthermore, the food material obtained by dispersing an
equol-containing fermented soybean hypocotyl or its extract into
cacao mass has reduced bitterness and exhibits excellent flavor,
although it contains an equol-containing fermented soybean
hypocotyl or its extract. The food material of the present
invention can be used in various foods without adversely affecting
the flavor. When conventional food material is added to a baked
confectionary in a granular form or a chip form, the components in
the food material tend to diffuse into the entire baked
confectionary in the baking step, and a desirable flavor may not be
obtained. In the food material of the present invention, the
equol-containing fermented soybean hypocotyl is covered with cacao
mass. Therefore, even when the food material of the present
invention is added to a baked confectionary in a granular form or a
chip form, diffusion of the effective components within the food
material can be suppressed and a desirable flavor can be
obtained.
[0093] The useful physiological benefits of the food of the present
invention can be enjoyed based on the useful physiological activity
of the equol-containing fermented soybean hypocotyl or its
extract.
[0094] The equol-containing fermented soybean hypocotyl or its
extract has reduced soybean hypocotyl-derived allergens, and
therefore the food material and various foods of the present
invention can be safely taken by or administered to people
suffering from soybean allergy.
BEST MODE FOR CARRYING OUT THE INVENTION
[0095] The following details the present invention.
1. Production Method for Equol-Containing Extract
[0096] The present invention provides a method for producing an
equol-containing extract using an equol-containing fermented
soybean hypocotyl as a material. The production method for the
equol-containing extract of the present invention is divided into
the following Production Methods I and II. The following details an
equol-containing fermented soybean hypocotyl used as a material of
the method of the present invention, and describes in detail
Production Methods I and II.
Equol-Containing Fermented Soybean Hypocotyl
[0097] In the production method for the equol-containing extract
according to the present invention, an equol-containing fermented
soybean hypocotyl is used as a raw material. The following
describes an equol-containing fermented soybean hypocotyl.
[0098] An equol-containing fermented soybean hypocotyl is a kind of
a fermented soybean hypocotyl produced by fermenting a soybean
hypocotyl using an equol-producing microorganism.
[0099] The equol-producing microorganism used for the production of
the equol-containing fermented soybean hypocotyl is selected from
microorganisms having the ability (metabolic activity) to produce
equol by metabolizing at least one daidzein compound selected from
the group consisting of daidzein glycosides, daidzein, and
dihydrodaidzein. Examples of the daidzein glycosides here include
daidzin, malonyldaidzin, and acetyldaidzin.
[0100] The equol-producing microorganism is not limited and may be
any microorganism having the foregoing ability. For example, any
publicly known microorganisms, or any microorganisms obtained by a
general screening method, may be used. It is known that such
equol-producing microorganisms exist in the microorganisms
belonging to Lactococcus, such as Lactococcus garvieae;
microorganisms belonging to Streptococcus, such as Streptococcus
intermedius or Streptococcus constellatus; or microorganisms
belonging to Bacteroides such as Bacteroides ovatus. Among various
kinds of equol-producing microorganism, the present invention
prefers lactic acid bacteria belonging to Lactococcus or
Streptococcus, further prefers lactic acid bacteria belonging to
Lactococcus, and in particular, lactic acid bacteria belonging to
Lactococcus garvieae. The equol-producing microorganisms may be
isolated from, for example, human feces, based on the index of the
existence of an equol-producing property. The inventors of the
present invention deposited some identified bacterium that had been
isolated from human feces; namely, Lactococcus 20-92 (FEM
BP-10036), Streptococcus E-23-17 (FEM BP-6436), Streptococcus
A6G225 (FEM BP-6437), and Bacteroides E-23-15 (FEM BP-6435), that
are available to be used as the equol-producing microorganisms.
Among these, the present invention particularly prefers Lactococcus
20-92.
[0101] The equol-containing fermented soybean hypocotyl is produced
using soybean hypocotyl as a fermenting material. The soybean
hypocotyl designates a part corresponding to the plumule or the
radicle on the germination of soybean, and is known to contain a
large amount of daidzein compounds such as daidzein glycosides or
daidzein. The soybean hypocotyl used in the present invention is
not limited by the producing district of soybean or whether
processed or unprocessed, unless the daidzein compounds inside are
not significantly lost. For example, the equol-containing fermented
soybean hypocotyl may be a raw hypocotyl, or may be one isolated
from heated, dried or steam-boiled soybean, or one obtained by
heating, drying or steam-boiling hypocotyl that is isolated from
unprocessed soybean. Further, the soybean hypocotyl may be
processed by degreasing or deproteinization. The form of the
soybean hypocotyl is not particularly limited, and may be in the
form of powder, chunks, or pulverized or fragmentized grains.
Powdery soybean hypocotyl is particularly preferable because of its
efficient equol production. The fermentation of soybean hypocotyl
is carried out by first adjusting the water content of the soybean
hypocotyl by adding an appropriate amount of water, and then
inoculating the equol-producing microorganism to the hypocotyl.
[0102] The amount of water added to the soybean hypocotyl is
adjusted depending on the type of equol-producing microorganism or
the type of fermenter. The ratio of water to the soybean hypocotyl
in the beginning of fermentation is 400 to 4,000 parts by weight,
preferably 500 to 2,000 parts by weight, further preferably 600 to
1,000 parts by weight, based on 100 parts by weight (dry weight) of
the soybean hypocotyl.
[0103] Further, if necessary, some additives may be added to the
soybean hypocotyl as the raw material in the fermentation to
improve the fermentation efficiency or flavor of the product.
Examples of the additives include nitrogen sources such as a yeast
extract, polypeptone, or a meat extract; carbon sources such as
glucose or sucrose; inorganic salts such as phosphate, carbonate or
hydrosulfate; vitamins; and nutritional components such as amino
acid. Particularly, when using an equol-producing microorganism for
converting arginine into ornithine (hereinafter referred to as an
"ornithine/equol-producing microorganism"), arginine is added to
the soybean hypocotyl before fermentation so that the fermented
substance contains ornithine. In this case, the amount of arginine
is, for example, about 0.5 to 3 parts by weight, based on 100 parts
by weight (dry weight) of the soybean hypocotyl. The
ornithine/equol-producing microorganism may be obtained by a
publicly-known screening method using an index of the
equol-producing ability and the conversion ability from arginine
into ornithine. The ornithine/equol-producing microorganism may be
selected from the group of Lactococcus garvieae, typified by
Lactococcus 20-92 (FEM BP-10036).
[0104] Further, the pH of the fermenting material (a soybean
hypocotyl containing substance) is not particularly limited within
a range for the equol-producing microorganism to grow; however, to
secure excellent proliferation of the equol-producing
microorganism, the pH of the fermenting material preferably falls
to within about 6 to 7, more preferably about 6.3 to 6.8.
[0105] Further, isoflavone containing the above-mentioned daidzein
compounds may be added to the fermenting material (a soybean
hypocotyl containing substance). Such inclusion of isoflavone in
the fermenting material increases the equol content of the
fermented soybean hypocotyl, thereby improving the usability of the
fermented soybean hypocotyl.
[0106] The fermentation of the soybean hypocotyl is carried out
under appropriate conditions according to the growing
characteristic of the equol-producing microorganisms. For example,
when the equol-producing microorganisms listed above are used, the
fermentation of the soybean hypocotyl is carried out under
anaerobic conditions.
[0107] The fermentation temperature is not limited within an
appropriate range for the equol-producing microorganism to grow.
The temperature is generally 20 to 40.degree. C., preferably 35 to
40.degree. C., and more preferably 36 to 38.degree. C.
[0108] The fermentation time is determined depending on the desired
production amount of equol, the residual amount of daidzein
compounds, the type of the equol-producing microorganism or the
like. The fermentation time is generally 1 to 10 days, preferably 2
to 7 days, and further preferably 3 to 5 days.
[0109] In the fermented soybean hypocotyl obtained through the
fermentation under the foregoing conditions, the produced equol is
accumulated, and thereby the useful physiologic activity of equol
can be expressed. The equol content in the fermented soybean
hypocotyl varies depending on the type of the equol-producing
microorganism or fermentation conditions. The equol content in the
fermented soybean hypocotyl is generally 1 to 20 mg, preferably 2
to 12 mg, and more preferably 5 to 8 mg, per gram (dry weight) of
the fermented soybean hypocotyl.
[0110] Apart from equol, the equol-containing fermented soybean
hypocotyl also contains various isoflavones such as daidzein
compounds including daidzin, malonyldaidzin, acetyldaidzin,
daidzein, or dihydrodaidzein (these components are referred to as
"daidzein compounds", hereinafter); genistein compounds including
malonylgenistin, acetylgenistin, genistein, dihydrogenistein (these
components are referred to as "genistein compounds", hereinafter);
or glycitein compounds including glycitin, malonylglycitin,
acetylglycitin, glycitein, dihydroglycitein (these components are
referred to as "glycitein compounds" hereafter). The isoflavones
also express the useful physiologic activities. The content of each
isoflavone (incl. equol) in the fermented soybean hypocotyl is
generally 5 to 20 mg, preferably 5 to 15 mg, and more preferably 8
to 15 mg, per gram (dry weight) of the equol-containing fermented
soybean hypocotyl.
[0111] Further, the equol-containing fermented soybean hypocotyl
also differs from general soybean hypocotyls in terms of the
composition of isoflavones other than equol. Particularly, the
total content of genistein compounds in the equol-containing
fermented soybean hypocotyl is preferably not more than 15 wt. %,
and more preferably not more than 12 wt. %. With such a low
genistein compound content, the equol-containing fermented soybean
hypocotyl is more advantageous than the unfermented soybean
hypocotyl in terms of the composition of isoflavones. Note that
genistein compounds are kinds of isoflavone and are chemicals
suspected of endocrine disruption, and there is a demand for the
reduction in genistein compounds concentrations in food (Young H J,
Jodi F, Kimberly F A, Daniel R D, William G H. Effects of dietary
daidzein and its metabolite, equol, at physiological concentrations
on the growth of estrogen-dependent human breast cancer (MCF-7)
tumors implanted in ovariectomized athymic mice. See Carcinogenesis
127(4):856-863, 2006).
[0112] The following is an isoflavone composition example of the
equol-containing fermented soybean hypocotyl (the unit "mg"
represents the total amount of each isoflavone per gram (dry
weight) of the fermented soybean hypocotyl). [0113] Equol: 1 to 20
mg, preferably 2 to 12 mg [0114] Daidzein compounds: 0.1 to 30 mg,
preferably 0.1 to 1.5 m [0115] Genistein compounds: 0.05 to 2.5 mg,
preferably 0.05 to 2 mg [0116] Glycitein compounds: 0.1 to 4 mg,
preferably 2 to 3.5 mg.
[0117] The following is a composition ratio example of the
isoflavones in the equol-containing fermented soybean hypocotyl
(the unit "wt. %" represents a ratio relative to the total amount
of isoflavones in the equol-containing fermented soybean
hypocotyl). [0118] Equol: 30 to 75 wt. %, preferably 40 to 70 wt.
%, more preferably 45 to 70 wt. % [0119] Daidzein compounds: 1 to
20 wt. %, preferably 2 to 15 wt. %, more preferably 4 to 12 wt. %
[0120] Genistein compounds: 0.1 to 20 wt. %, preferably 1 to 15 wt.
%, more preferably 1 to 10 wt. % [0121] Glycitein compounds: 10 to
50 wt. %, preferably 15 to 35 wt. %, more preferably 25 to 35 wt.
%.
[0122] Since this equol-containing fermented soybean hypocotyl has
a particular isoflavone composition that is not found in any prior
art, it can be defined as an isoflavone-containing substance
specified by the foregoing isoflavone composition.
[0123] Lactococcus 20-92 (FEM BP-10036) is particularly preferred
as an equol-producing microorganism in the production of the
equol-containing fermented soybean hypocotyl having the foregoing
isoflavone composition.
[0124] The equol-containing fermented soybean hypocotyl also
contains saponin derived from the soybean hypocotyl. The
equol-containing fermented soybean hypocotyl generally contains
saponin in an amount of 10 to 80 mg, preferably 20 to 50 mg, and
more preferably 30 to 40 mg, per gram (in dry weight) of the
equol-containing fermented soybean hypocotyl.
[0125] Further, as described above, ornithine is contained in the
fermented soybean hypocotyl produced from fermentation with an
ornithine/equol-producing microorganism after adding arginine to
soybean hypocotyl. The ornithine content of the equol-containing
fermented soybean hypocotyl is 5 to 20 mg, preferably 8 to 15 mg,
and more preferably 9 to 12 mg, per gram (dry weight) of the
equol-containing fermented soybean hypocotyl.
[0126] The equol-containing fermented soybean hypocotyl obtained
through fermentation under the foregoing conditions may be used
directly after fermentation as a material of the production method
of the present invention, or may be dried after fermentation to be
a dry solid before being used as the material of the production
method of the present invention.
[0127] Further, the equol-containing fermented soybean hypocotyl
obtained through fermentation under the foregoing conditions may be
subjected to preprocessing, such as defatting, before the following
Step I-1. The defatting can be performed by a method using hexane
as an extractant.
1-1. Production Method I
[0128] In Production Method I, the production of the
equol-containing extract is accomplished by carrying out Step I-1,
which is described below.
Step I-1
[0129] Production Method I for producing the equol-containing
extract comprises a Step I-1 of extracting an equol-containing
fermented soybean hypocotyl using an aqueous ethanol solution as an
extractant, and collecting liquid extract.
[0130] The ethanol concentration of the aqueous ethanol solution (a
mixed solution of ethanol and water) used in Step I-1 is generally
20 to 98 vol. %, preferably 45 to 95 vol. %, and more preferably 65
to 85 vol. %. By meeting this concentration range, it becomes
possible to extract useful components including equol from the
equol-containing fermented soybean hypocotyl.
[0131] The extraction in Step I-1 uses an aqueous ethanol solution
in an amount of, for example, 1 to 1000 parts by weight, preferably
2 to 500 parts by weight, more preferably 5 to 20 parts by weight,
per part by weight (dry weight) of the equol-containing fermented
soybean hypocotyl.
[0132] The Step I-1 extraction conditions are not particularly
limited, and a warm or cold dipping extraction method, percolation
method or the like may be used. More specifically, an aqueous
ethanol solution is added to the equol-containing fermented soybean
hypocotyl, and the mixture is allowed to stand or stirred for at
least one minute at room temperature, preferably for 5 minutes to
48 hours at room temperature.
[0133] After such extraction, the liquid extract is isolated to
obtain an equol-containing extract. The collection of the liquid
extract is performed by a publicly-known method, such as filtration
or centrifugation.
[0134] The liquid extract thus isolated in Step I-1 contains useful
components including equol and glyciteins, and the amount of oil is
less than that in the equol-containing fermented soybean hypocotyl.
The liquid extract thus obtained in Step I-1 may be directly
incorporated into a food or medicine as a liquid equol-containing
extract, or may be solidified by removing the extractant before
incorporated into a food or medicine. Further, the liquid extract
thus obtained in Step I-1, either in the form of a liquid or a
solid equol-containing extract, may be subjected to equol
refining.
[0135] The liquid extract isolated in Step I-1 contains equol,
glyciteins, and saponin, which can cause unpleasant astringency,
bitterness, or acridness. It is thus preferable to subject the
liquid extract isolated in Step I-1 to the following Step I-2 so as
to remove saponin and increase the concentration of the useful
components, including equol.
Step I-2
[0136] Step I-2 comprises removing the extractant from the liquid
extract obtained in Step I-1, carrying out another extraction using
ethanol as an extractant, and collecting an extract.
[0137] In Step I-2, the removal of the extractant from the extract
obtained in Step I-1 can be performed by a publicly-known method
such as drying.
[0138] The solid obtained by the removal of the extractant from the
extract obtained in Step I-1 (this solid may be hereinafter
referred to as "Solid Material") is subjected to extraction using
ethanol as an extractant. The ethanol used as an extractant in Step
I-2 may be water-free ethanol that however may inevitably contain
vapor.
[0139] The extraction in Step I-2 uses, for example, 1-1000 parts
by weight, preferably 5-500 parts by weight, and more preferably
5-20 parts by weight of ethanol, per part by weight (dry weight) of
the solid material.
[0140] The condition in Step I-2 extraction is not particularly
limited, and may be performed by a warm or cold dipping extraction
method, percolation method or the like. More specifically, ethanol
is added to the solid material and the mixture is allowed to stand
or stirred for at least one minute at room temperature, preferably
for 5 minutes to 48 hours at room temperature. The Step I-2
extraction may be combined with supersonic treatment to increase
the equol collection rate.
[0141] After such extraction, the extract is isolated to obtain an
equol-containing extract. The collection of the extract is
performed by a publicly-known method, such as filtration or
centrifugation.
[0142] The extract thus isolated in Step I-2 contains a greater
amount of useful components including equol and glyciteins, and a
reduced amount of saponin. The usefulness of this extract is
therefore greater than the extract obtained in Step I-1. The
extract thus obtained in Step I-2 may be directly incorporated into
a food or medicine as a liquid equol-containing extract, or may be
solidified by removing the extractant before being incorporated
into a food or medicine. The extract thus obtained in Step I-2,
either in the form of a liquid or a solid equol-containing extract,
may be subjected to equol refining. Note that, in order to improve
the conservation stability of the equol-containing extract, it is
more preferable to solidify the liquid extract by removing the
extractant.
Composition and Use of Equol-Containing Extract Obtained by
Production Method I
[0143] The equol content of the liquid extract obtained in Step I-1
or Step I-2 is 0.2 to 40 wt. %, preferably 0.4 to 24 wt. %, and
more preferably 1 to 16 wt. %, based on the total amount of a dry
solid product (100 wt. %). The glycitein content of the liquid
extract obtained in Step I-1 or Step I-2 is 0.01 to 8 wt. %,
preferably 0.08 to 5 wt. %, and more preferably 0.2 to 3 wt. %,
based on the total amount of a dry solid product.
[0144] The equol-containing extract obtained in Step I-1 or Step
I-2 contains equol at a high concentration, and expresses the
useful activities based on equol. More specifically, the
equol-containing extract obtained in Step I-1 or Step I-2 has an
effect of preventing or treating various diseases or symptoms
including menopausal disorders, osteoporosis, prostatic
hypertrophy, and metabolic syndrome; or for blood cholesterol level
reduction, skin-whitening, pimples, intestinal control, obesity,
and diuresis. Since the useful activities are thus expressed, the
equol-containing extract obtained in Step I-1 or Step I-2 may be
incorporated into food or medicines.
[0145] When used for a foodstuff, the equol-containing extract
obtained in Step I-1 or Step I-2 is often incorporated into
beverages, granules, fine granules, capsules, tablets, powder,
dairy products, gums, gummy candies, puddings, bars, and other
various dry foods. The food containing the equol-containing extract
may be used as a general food product, and also as a specified
health food, nutritional supplement, functional food, invalid food
or the like. Particularly, any food containing the fermented
soybean hypocotyl of the present invention is useful as a
nutritional supplement. Note that the details of foodstuffs
containing the equol-containing extracts obtained in Step I-1 or
Step I-2 are described later.
[0146] When the equol-containing extract obtained in Step I-1 or
Step I-2 is incorporated into a foodstuff, the proportion of the
equol-containing extract in the food varies depending on the type
of food, the equol content, the age and sex of the consumer, the
desired effect etc. Typically, the total amount (dry weight base)
of the equol-containing extract is 0.1 to 90 g, per 100 g of
food.
[0147] When used for a medicinal product, the equol-containing
extract obtained in Step I-1 or Step I-2 is often incorporated into
tablets, pills, powder, liquids, suspensions, emulsions, granules,
capsules, or suppositories. Such a medicinal product containing the
equol-containing extract has an effect of preventing or treating
various diseases or symptoms including menopausal disorders
(indefinite menopausal complaints, osteoporosis, hyperlipidemia),
osteoporosis, prostatic hypertrophy, and metabolic syndrome; or for
blood cholesterol level reduction, skin-whitening, pimples,
intestinal control, obesity, and diuresis. Particularly, the
medicinal product containing the equol-containing extract is
suitable for the prevention or treatment of indefinite complaint or
postmenopausal symptoms (e.g. osteoporosis, menopausal disorder) of
middle-aged women.
[0148] The dose of the medicinal product containing the
equol-containing extract obtained in Step I-1 or Step I-2 cannot be
uniformly defined, as it varies depending on the equol content in
the equol-containing extract, age, weight, symptom of the object or
the dosage number. The adult dosage per day typically corresponds
to the amount (dry weight) of the equol-containing extract from 1
to 2000 mg, preferably 5 to 1000 mg, and more preferably 40 to 50
mg. It is particularly preferable to determine the dose of the
equol-containing extract so that 2 to 30 mg of equol is
administered per day.
1-2. Production Method II
[0149] Production Method II produces an equol-containing liquid
extract by carrying out the following Step II-1 and Step II-2. The
following details Production Method II.
Step II-1
[0150] Production Method II first suspends an equol-containing
fermented soybean hypocotyl in water, adds an acid to the
suspension to adjust the pH to a range of 2 to 6, and isolates an
insoluble residue (Step II-1). In other words, in Step II-1, an
equol-containing fermented soybean hypocotyl is suspended in an
aqueous solution (pH 2 to 6) containing an acid, thereafter
collecting an insoluble residue.
[0151] In Step II-1, an equol-containing fermented soybean
hypocotyl is suspended in water, and an acid is added to the
suspension to adjust the pH to a range of from 2 to 6.
[0152] The acid used in Step II-1 may be an organic acid or an
inorganic acid. The organic acid is not limited as long as it is
adjustable to the foregoing pH range. Examples of the organic acid
include citric acids, L-tartaric acids, fumaric acids, L-ascorbic
acids, gluconic acids, acetic acids, lactic acids, DL-malic acids,
oxalic acids, formic acids, propionic acids, butanoic acids, and
methanesulfone acids. Similarly, the inorganic acid is not limited
as long as it is adjustable to the foregoing pH range. Examples of
the inorganic acid include hydrochloric acids, sulfuric acids,
nitric acids and phosphoric acids. Among them, acetic acids,
hydrochloric acids and sulfuric acids are preferred.
[0153] In Step II-1, the amount of water in which the
equol-containing fermented soybean hypocotyl is suspended is, for
example, 3 to 30 parts by weight, more preferably 5 to 20 parts by
weight, per part by weight (dry weight) of an equol-containing
fermented soybean hypocotyl.
[0154] In Step II-1, an equol-containing fermented soybean
hypocotyl is mixed in an aqueous solution 2 to 6 in pH, and the
solution is stirred or allowed to stand at room temperature for at
least 30 minutes, more preferably 2 to 24 hours.
[0155] Such treatment leaves equol as a part of the insoluble
residue, and the equol can be obtained by collecting the insoluble
residue using a publicly-known method such as filtration or
centrifugation.
[0156] The insoluble residue thus obtained in Step II-1 is then
subjected to the following Step II-2.
Step II-2
[0157] The insoluble residue thus obtained in Step II-1 is
subjected to another extraction using an aqueous ethanol solution
as an extractant. The resulting liquid extract is isolated (Step
II-2).
[0158] The insoluble residue obtained in Step II-1 may be subjected
to Step II-2, regardless of whether it is a dry product or a wet
product. However, when the insoluble residue is a wet product, it
is preferable to adjust the water content of the aqueous ethanol
solution (described later) in consideration of the measured water
content of the insoluble matter.
[0159] The ethanol concentration of the aqueous ethanol solution (a
mixed solution of ethanol and water) used in Step II-2 is, for
example, 50 to 95 vol. %, more preferably 70 to 90 vol. %. With
this concentration range, the equol extraction obtained from the
insoluble residue in Step II-2 can be efficiently performed.
[0160] The extraction in Step II-2 uses an aqueous ethanol solution
in an amount of, for example, 2 to 30 parts by weight, more
preferably 4 to 10 parts by weight, per part by weight (dry weight)
of the insoluble residue obtained in Step II-1.
[0161] The condition in the extraction in Step II-2 is not
particularly limited, and the warm or cold dipping extraction
method, percolation method or the like may be used. More
specifically, an aqueous ethanol solution is added to the insoluble
residue obtained in Step II-1, and the mixture is allowed to stand
or stirred for at least 30 minutes at room temperature, preferably
1 minute to 50 hours at room temperature. After such extraction,
the liquid extract is isolated to obtain an equol-containing
fraction. The collection of the extract is performed by a
publicly-known method such as filtration or centrifugation.
[0162] Because of the inclusion of concentrated equol, the liquid
extract thus isolated in the Step II-2 contains equol at a high
concentration and useful components such as glysitein compounds,
the extract is more useful than the equol-containing fermented
soybean hypocotyl. The liquid extract obtained in Step II-2 may be
directly incorporated into a food or medicine as a liquid
equol-containing extract, or may be processed into powder by
removing the extractant before incorporated into a food or
medicine. The liquid extract thus obtained in Step II-2, either in
the form of a liquid or a solid equol-containing extract, may be
subjected to equol refining. Note that, in order to improve the
conservation stability of the equol-containing extract, it is more
preferable to solidify the liquid extract by removing the
extractant.
[0163] The removal of the extractant from the liquid extract
obtained in Step II-2 may be performed under the same conditions as
in Step I-2 of Production Method 1; however, the removal is
preferably performed by spray-drying.
Composition and Use of Equol-Containing Extract Obtained by
Production Method II
[0164] By drying the liquid extract obtained in Method II (the
liquid extract obtained in the foregoing Step II-2), it is possible
to isolate equol with a collection rate of 90% or greater. Thus
obtained is equol-containing powder that contains equol ten times
greater than the equol in an equol-containing fermented soybean
hypocotyl. With this high equol content, the equol-containing
powder more effectively expresses the useful activities derived
from equol. The purpose or method of use of the equol-containing
powder obtained in Method II is the same as that of the
equol-containing powder obtained in Method I.
2. Equol Purification Method
[0165] The present invention also provides a method for purifying
an equol-containing substance to obtain equol. The equol
purification method of the present invention is divided into the
following Purification Methods 1 and 2. The following details an
equol-containing fermented soybean hypocotyl to be refined by the
purification method of the present invention and describes in
detail Purification Methods 1 and 2.
Equol-Containing Substance
[0166] In the purification method of the present invention, the
equol-containing substance to be subjected to the purification
process is not limited as long as it contains equol. For example,
the equol-containing substance used for the method may be a
reaction product containing equol generated by chemical synthesis,
or an equol-containing fermented substance produced by
fermentation. The equol-containing fermented substance obtained
through fermentation contains many kinds of isoflavone similar in
structure to equol. However, since the method of the present
invention can advantageously separate equol from isoflavone in a
efficient manner, an equol-containing fermented substance is
suitable for an equol-containing substance for the purification
method of the present invention.
[0167] The following explains an equol-containing fermented
substance.
[0168] The equol-containing fermented substance was produced
through a publicly-known fermentation method using an
equol-producing microorganism. More specifically, a microorganism
with the ability (metabolic activity) to produce equol by
metabolizing at least one daidzein compound selected from the group
consisting of daidzein glycosides, daidzein, and dihydrodaidzein,
is inoculated in a fermenting material (material to be subjected to
the fermentation) containing the daidzeins. The sample is then
fermented (cultured) under the conditions suitable to grow the
microorganism. The resulting fermented substance contains
equol.
[0169] The equol-producing microorganism is selected from the list
of "Equol-Containing Fermented Soybean Hypocotyl" in the section
"1. Production Method for Equol-Containing Extract".
[0170] The fermenting material containing daidzein compounds is not
limited by other factors; however, the material is preferably
approved for its safety as a food material. Examples of the
fermenting material containing daidzein compounds include soybeans,
soybean hypocotyl, soybean hypocotyl extract, tofu, deep-fried
tofu, soy milk, fermented soybeans, soy sauce, bean paste, a
tempeh, and a red clove or its extract, alfalfa or its extract. A
suitable fermenting material containing daidzein compounds is a
soybean hypocotyl because of its high daidzein content.
[0171] Further, an isoflavone containing any of above-described
daidzein compound may be added in advance to the fermenting
material containing daidzein compounds. With this extra addition of
isoflavone to the fermenting material, the equol content of the
resulting fermented substance increases.
[0172] Further, as required, some additives may be used in the
fermenting material containing daidzein compounds to improve
fermentation efficiency etc. Examples of the additives include
nitrogen sources such as a yeast extract, poly peptone, or a meat
extract; carbon sources such as glucose or sucrose; inorganic salts
such as phosphate, carbonate or hydrosulfate; vitamins; and
nutritional components such as amino acid.
[0173] Various fermentation conditions including water content,
time, temperature, atmosphere in the production of an
equol-containing fermented substance are determined depending on
the type of the equol-producing microorganism, the type of the
fermenting material, the production amount of equol, the residual
daidzein compound amount, or the like.
[0174] For the equol-containing substance used for the purification
method of the present invention, the equol-containing fermented
soybean hypocotyl described in the section "1. Production Method
for Equol-Containing Extract" is particularly preferable.
2-1. Purification Method 1
[0175] The Purification Method 1 refines equol by carrying out the
following Steps 1-1 to 1-5.
Step 1-1
[0176] The purification method of the present invention carries out
extraction with respect to an equol-containing substance using an
aqueous ethanol solution as an extractant, and then collects the
resulting liquid extract (Step 1-1).
[0177] A typical ethanol concentration of the aqueous ethanol
solution used in Step 1-1 is 20 to 98 vol. %, preferably 45 to 95
vol. %, and more preferably 45 to 60 vol. %. Ensuring this ethanol
concentration, it is possible to efficiently extract equol from the
equol-containing substance.
[0178] The extraction in Step 1-1 uses an aqueous ethanol solution
in an amount of, for example, 1 to 1000 parts by weight, preferably
2 to 500 parts by weight, more preferably 5 to 20 parts by weight,
per part by weight (dry weight) of the equol-containing
substance.
[0179] The conditions in the extraction in Step 1-1 are not
particularly limited, and a warm or cold dipping extraction method,
percolation method or the like may be used. More specifically, an
aqueous ethanol solution is added to the equol-containing
substance, and the mixture is allowed to stand or stirred for at
least one minute at room temperature, more preferably 5 minutes to
48 hours at room temperature.
[0180] After such extraction, the liquid extract is isolated to
obtain an equol-containing fraction. The collection of the liquid
extract is performed by a publicly-known method such as filtration
or centrifugation.
[0181] The liquid extract thus isolated in Step 1-1 contains a
reduced amount of oil in the equol-containing substance, and
isoflavone including glycitein compounds and saponin, as well as
equol.
[0182] The liquid extract thus obtained in Step 1-1 is subjected to
the following Step 1-2.
Step 1-2
[0183] After removing the extractant from the liquid extract
obtained in Step 1-1, the residue is subjected to extraction using
ethanol as an extractant, and the liquid extract is isolated (Step
1-2).
[0184] In Step 1-2, the removal of the extractant from the liquid
extract obtained in Step 1-1, and another extraction of the
thus-obtained residue using an ethanol, and the collection of the
liquid extract are carried out in the same conditions as those of
Step I-2 of Production Method I in the section "1. Production
Method for Equol-Containing Extract".
[0185] The liquid extract collected in Step 1-2 contains a reduced
amount of saponin, and contains isoflavone including glycitein
compounds, as well as equol. The liquid extract thus obtained in
Step 1-2 is subjected to the following Step 1-3.
Step 1-3
[0186] After removing the extractant from the liquid extract
obtained in Step 1-2, the residue is suspended in hexane to isolate
an insoluble matter (Step 1-3).
[0187] In Step 1-3, the removal of extractant from the liquid
extract obtained in Step 1-2 may be performed by a publicly-known
method such as distillation under reduced pressure, drying or the
like.
[0188] The extractant is removed from the liquid extract obtained
in Step 1-2, and hexane is added to the residue (hereinafter
occasionally referred to as "second material residue") to suspend
the residue in hexane.
[0189] The extraction in Step 1-3 uses hexane in an amount of 1 to
50 parts by weight, and more preferably 5-10 parts by weight, per
part by weight (dry weight) of the second material residue.
[0190] The conditions for the extraction in Step 1-3 are not
particularly limited. For example, a warm or cold dipping
extraction method may be adopted. More specifically, hexane is
added to the second material residue, and the mixture is allowed to
stand or stirred for at least 5 minute at room temperature,
preferably 3 minutes to an hour at room temperature.
[0191] After such suspension using hexane, equol is extracted as an
insoluble matter. Therefore, by collecting the insoluble matter, an
equol-containing extract is obtained. The collection of the
insoluble residue is performed by a publicly-known method such as
filtration or centrifugation.
[0192] Through Step 1-3, lipid-soluble impurities are removed from
the liquid extract obtained in Step 1-2. The insoluble residue
obtained in Step 1-3 is subjected to the following Step 1-4.
Step 1-4
[0193] Step 1-4 comprises subjecting the insoluble residues
obtained in Step 1-3 to extraction using a mixed solution of hexane
and ether as an extractant, and collecting the resulting liquid
extract (Step 1-4).
[0194] The ether, as one of the components of the extractant used
in Step 1-4, is not particularly specified. Examples of the ether
include dimethylether, methylethylether, and diethylether, and they
may be used singly or in combination. Among them, diethylether is
particularly preferred because of its equol refining
efficiency.
[0195] The mixture ratio of the mixed solution of hexane and ether
used in Step 1-4 is not particularly limited; however, to obtain
equol of desired purity, the ratio of hexane to ether in volume is
10:90 to 20:80, and preferably 30:70 to 25:75.
[0196] The extraction in Step 1-4 uses the mixed solution of hexane
and ether in an amount of 50 to 5000 parts by weight, more
preferably 200 to 1000 parts by weight, per part by weight (dry
weight) of the insoluble residue obtained in Step 1-3.
[0197] The conditions for the extraction in Step 1-4 are not
particularly limited. For example, a warm or cold dipping
extraction method or percolation method may be adopted. More
specifically, the mixed solution of hexane and ether is added to
the insoluble matter obtained in Step 1-3, and the mixture is
allowed to stand or stirred for at least 10 minutes at room
temperature, preferably 15 minutes to 30 hours at room
temperature.
[0198] After such extraction, the liquid extract is isolated to
obtain equol. The collection of the liquid extract is performed by
a publicly-known method such as filtration or centrifugation.
Step 1-5
[0199] The liquid extract obtained in Step 1-4 is subjected to
silica gel chromatography to obtain an equol-containing fraction
(Step 1-5).
[0200] Step 1-5 may adopt any kind of silica gel chromatography
that uses separate columns each containing silica gel as a filler.
Examples of silica gel column chromatography include open column
chromatography that causes a solvent flow by gravity drop, flash
column chromatography that makes a solvent flow using a pump, and
high-performance liquid chromatography. Open column chromatography
and flash column chromatography are particularly preferred, as they
can be easily carried out in an industrial manner. When carrying
out silica gel column chromatography, it is preferable to use a
mixed solution of hexane and ether as a mobile phase (developing
solvent) to further improve the equol refining degree. The ether
used for the mobile phase is not limited. Examples of the ether
include dimethylether, methylether, and diethylether, and they may
be used solely or in combination. To improve the equol refining
degree, the ether is preferably the same as that used for the mixed
solution of hexane and ether in Step 1-4. In the elution of equol
using silica gel column chromatography in Step 1-5, it is
preferable to use a mixed solution of hexane and ether identical in
composition to that used in Step 1-4 as the initial mobile phase,
and the ratio of ether is gradually increased according to the
gradient method. Such an increase of the ratio of ether in the
mobile phase causes the equol to separate from other impurities,
together with the mobile phase in which hexane and ether is mixed
at 40:60 to 20:80, more preferably 30:70 to 25:50. Thereafter, the
fraction containing equol having been eluted by silica gel column
chromatography is isolated to obtain highly pure equol. Then, the
extractant is removed from the equol-containing fraction by a
publicly-known method such as distillation under reduced pressure,
drying or the like. This removes most of the impurities from the
fraction, thereby leaving a highly pure equol solid.
2-2. Prification Method 2
Step 2-1
[0201] Purification method 2 of the present invention carries out
extraction with respect to an equol-containing substance using, as
an extractant, an organic solvent or an aqueous organic solvent of
at least one member selected from the group consisting of ethyl
acetate, alcohol, acetone, dioxane, acetonitrile, diethylether, and
toluene, and then concentrates the liquid extract (Step 2-1).
[0202] In Step 2-1, the extractant may be an organic solvent of at
least one member selected from the group consisting of ethyl
acetate, alcohol, acetone, dioxane, acetonitrile, diethylether, and
toluene. However, to ensure efficient equol extraction, it is more
preferable to use an aqueous organic solvent that is obtained by
adding water to the aforementioned organic solvent.
[0203] Examples of the alcohol used as an extractant include
methanol, ethanol, isopropyl alcohol, and n-propyl alcohol.
[0204] A typical concentration of the aqueous organic solvent is 80
to 99 vol. %, preferably 90 to 97 vol. %. Ensuring this
concentration, it is possible to obtain a liquid extract with a
high equol content.
[0205] One preferable extractant used in Step 2-1 is aqueous ethyl
acetate.
[0206] The extraction conditions are not particularly limited, and
a warm or cold dipping extraction method or the like may be used.
More specifically, an extractant is added to the equol-containing
substance, and the mixture is allowed to stand or stirred for at
least 30 minutes at room temperature, more preferably for 1 hour to
50 hours at room temperature.
[0207] The conditions in the Step 1-1 extraction are also not
particularly limited, and a warm or cold dipping extraction method,
percolation method or the like may be used.
[0208] After such extraction, equol educed in the liquid extract is
isolated by a publicly-known solid-liquid dividing method such as
filtration or centrifugation. The obtained liquid extract is then
concentrated, for example, under reduced pressure to obtain an
equol concentrate. The equol concentrate thus obtained is subjected
to the following Step 2-2.
Step 2-2
[0209] The equol concentrate obtained in Step 2-1 is subjected to
silica gel column chromatography to obtain an equol-containing
fraction (Step 2-2).
[0210] In Step 2-2, the equol concentrate obtained in Step 2-1 may
be directly subjected to silica gel column chromatography.
[0211] Further, if the extraction in Step 2-1 is carried out with
an aqueous solvent containing alcohol, an equivalent amount of
ethyl acetate and water that is 5 to 10 times in volume the
concentrated liquid extract are added to the concentrate, and the
ethyl acetate fraction is separated and concentrated to be
subjected to column chromatography. This column chromatography in
Step 2-2 may be any kind of chromatography that uses separate
columns each containing silica gel as a filler. Examples of silica
gel column chromatography include open column chromatography that
causes a solvent flow by gravity drop, flash column chromatography
that makes a solvent flow using a pump, and high-performance liquid
chromatography. Open column chromatography and flash column
chromatography are particularly preferred, as they can be easily
carried out in an industrial manner.
[0212] In the silica gel column chromatography in Step 2-2, it is
preferable to use a mixed solution of an ethyl ether or ethyl
acetate solution (Solution A, hereinafter) and an n-hexane,
petroleum ether or n-heptane solution (Solution B, hereinafter) as
a mobile phase. The ratio of Solution B in the mixed solution is 2
to 50 parts by volume, more preferably 3 to 10 parts by volume, per
part by volume of Solution A.
[0213] The liquid extract obtained in Step 2-1 is thus subjected to
silica gel column chromatography to isolate the equol-containing
fraction.
[0214] The equol-containing fraction obtained in Step 2-2 is
subjected to Step 2-3
Step 2-3
[0215] The solvent is removed from the equol-containing fraction
obtained in Step 2-2, and the resulting residue is subjected to
recrystallization. The precipitated crystals are then isolated
(Step 2-3).
[0216] The removal of a solvent from the equol-containing fraction
obtained in Step 2-2 may be performed by a publicly-known solvent
removal method, such as concentration under reduced pressure.
[0217] The solvent used for the recrystallization in Step 2-3 is
not particularly limited as long as it can be used for the
recrystallization of equol. A typical solvent is a mixed solution
of ethanol and water (aqueous ethanol solution), ethyl acetate, a
mixed solution of ethyl acetate and hexane, or a mixed solution of
ethanol and hexane. The mixed solution of ethyl acetate and hexane
and the aqueous ethanol solution are particularly preferable. More
specifically, the mixed solution of ethyl acetate and hexane
typically contains ethyl acetate and hexane at a volume ratio of
10:1 to 1:10. The ethanol concentration of the aqueous ethanol
solution used for the recrystallization is typically 30 to 90 vol.
%, preferably 40 to 70 vol. %. By carrying out the
recrystallization using the mixed solution of ethyl acetate and
hexane or an aqueous ethanol solution adjusted to the foregoing
concentration, the purity of the resultant equol further
increases.
[0218] The recrystallization in Step 2-3 can be carried out using a
general method. For example, a solvent for recrystallization is
added to the residue resulted from the removal of solvent from the
equol-containing fraction obtained in Step 2-2, and the residue is
dissolved in the solvent by heating. The solution is then cooled to
precipitate equol crystals. To further facilitate or accelerate
equol crystallization, equol crystal seeds may be inoculated in the
sample.
[0219] The temperature condition in the crystallization is
determined according to the boiling point and the freezing point of
the solvent. Generally, the residue is dissolved at about the
boiling point of the solvent, and crystals are precipitated at
about 0 to 25.degree. C.
3. Food Material
[0220] The present invention further provides a food material in
which the equol-containing fermented soybean hypocotyl or its
extract is dispersed in cacao mass. The following describes the
details, such as the ingredients, of the food material of the
present invention.
Equol-Containing Fermented Soybean Hypocotyl and Extraction
Thereof
[0221] The equol-containing fermented soybean hypocotyl used for
the food material of the present invention is the same as the
equol-containing fermented soybean hypocotyl described above in the
section "1. Production Method for Equol-Containing Extract". The
equol-containing fermented soybean hypocotyl may be used for the
food material of the present invention immediately after the
fermentation, or after dried into a dry solid as required. To
improve the conservation stability of the equol-containing
fermented soybean hypocotyl, it is more preferable to solidify the
equol-containing fermented soybean hypocotyl by heated air-drying
or the like. The air-dried equol-containing fermented soybean
hypocotyl may be further processed into powder as required.
[0222] The extract of the equol-containing fermented soybean
hypocotyl designates an equol-containing extract resulted from
solvent extraction with respect to the equol-containing fermented
soybean hypocotyl.
[0223] The extract of the equol-containing fermented soybean
hypocotyl is not particularly limited; however, the extract is
preferably the equol-containing extract obtained by the method
described above in the section "1. Production Method for
Equol-Containing Extract", more preferably the equol-containing
extract obtained by Production Method I described above. The
extract of the equol-containing fermented soybean hypocotyl of the
present invention may be used in a liquid state; however, it is
more preferable to solidify the liquid extract by removing the
extractant.
[0224] The food material of the present invention may contain
either of the equol-containing fermented soybean hypocotyl or the
extract thereof; however, the equol-containing fermented soybean
hypocotyl is nutritionally preferred.
Cacao Mass
[0225] Cacao mass is a food material mainly used as an ingredient
of chocolate. The cacao mass is obtained by processing cacao beans
with fermentation, torrefaction, and grinding. The cacao mass used
as a food material of the present invention is not particularly
limited, as long as its safety as a food material is approved.
Proportion of Equol-Containing Fermented Soybean Hypocotyl or
Extract Thereof and Cacao Mass
[0226] In the food material of the present invention, the
proportion of the equol-containing fermented soybean hypocotyl or
extract thereof and cacao mass is not limited; however, in order to
improve the flavor of the food more preferably, the proportion of
cacao mass is generally 10 to 2000 parts by weight (dry weight),
preferably 20 to 900 parts by weight, more preferably 20 to 400
parts by weight, based on 100 parts by weight of the
equol-containing fermented soybean hypocotyl or extract thereof.
Particularly, when using the equol-containing fermented soybean
hypocotyl, the proportion of cacao mass is generally 100 to 2000
parts by weight (dry weight), preferably 200 to 900 parts by
weight, and more preferably 200 to 400 parts by weight, based on
100 parts by weight of the equol-containing fermented soybean
hypocotyl or extract thereof.
[0227] The food material of the present invention may consist only
of the equol-containing fermented soybean hypocotyl or extract
thereof and cacao mass, but may also contain other ingredients as
detailed below. The sum proportion of the equol-containing
fermented soybean hypocotyl or extract thereof and cacao mass is 40
to 100 wt. %, preferably 60 to 95 wt. %, and more preferably 70 to
95 wt. %, based on the gross amount of the food material of the
present invention. By thus adjusting the proportion of the
ingredients, the useful bioactivity derived from the
equol-containing fermented soybean hypocotyl can be effectively
expressed, and the food will have a good flavor.
Arbitrary Ingredients
[0228] Insofar as the effects of the invention are not impaired,
the food material of the present invention may contain various
ingredients in addition to the (a) equol-containing fermented
soybean hypocotyl or extract thereof, and (b) cacao mass.
[0229] Examples of suitable ingredients for the food material of
the present invention include sweeteners such as sugar, trehalose,
acesulfam K, xylitol, scralose, erythritol, aspartame and starch
syrup; oils and fats such as cacao butter or vegetable oils;
emulsifiers such as soybean lecithin; perfume; perfume
preparations; milk; and water.
Form of Food Material
[0230] The shape of the food material of the present invention is
not particularly limited, and may be a form of powder, granule,
chip, or plate, depending on the target product form of the
food.
[0231] To make good use of the texture characteristic of the food
material of the present invention in the product, the food material
is preferably processed in the form of a granule or a chip,
particularly preferably a granule or a chip of about 30 to 1000 mg
per piece.
Production Method for Food Material of the Present Invention
[0232] The food material of the present invention is prepared by
mixing the equol-containing fermented soybean hypocotyl or extract
thereof, cacao mass, and other necessary ingredients at a
temperature where the cacao butter in the cacao mass dissolves
(preferably 30 to 50.degree. C.), and then cooling and solidifying
the mixture. As a result, the food material of the present
invention in which the equol-containing fermented soybean hypocotyl
is dispersed in cacao mass is prepared. As required, the food
material of the present invention is subjected to pulverization,
trituration, sizing or the like to be formed into a desired shape.
Alternatively, the food material of the present invention may be
shaped using extrusion granulation during the production.
Use of Food Material
[0233] The food material of the present invention may be
incorporated into various kinds of foodstuffs, as a raw material of
an equol-containing food, or as a food additive. The present
invention also provides such equol-containing foods made of the
above-described food material.
[0234] The equol-containing food of the present invention is not
particularly limited, and includes baked confectionaries and frozen
desserts.
[0235] Examples of the baked confectionaries include various
glutinous rice crackers; wheat crackers such as wafers or waffles;
biscuits; cookies; soda crackers; pies; cakes; doughnuts; and
bread. Examples of the frozen desserts include ice cream, shaved
ice, and ice pops.
[0236] Among them, baked confectionaries are one of the suitable
forms of equol-containing food. Particularly, a baked confectionary
made of soybean powder contains not only equol but also other
soybean-derived useful components; therefore, the baked
confectionary is highly useful, providing a superior health
maintenance effect.
[0237] One preferred example of the baked confectionary made of
soybean powder contains soybean powder, egg, and butter, but no
wheat. The soybean powder to be used for this type of cookie is
preferably processed by heating or the like to deactivate or
diminish the lipoxygenases and to remove or reduce the soybean
smell. Such a baked confectionary can be produced by, for example,
using 0 to 35 parts by weight of egg, 10 to 65 parts by weight of
butter, 0.1 to 50 parts by weight of the food material of the
present invention, and other ingredients, per 100 parts by weight
of soybean powder. These ingredients are mixed with an appropriate
amount of water to make the dough. The dough is then formed into a
predetermined shape, and baked at about 60 to 250.degree. C. for 2
to 60 minutes.
[0238] The proportion of the food material of the present invention
in the equol-containing food is not particularly limited, and is
adjusted depending on the equol content in the food material of the
present invention, the form of the equol-containing food or the
like. The proportion of the food material of the present invention
is generally 3 to 30 wt. %, preferably 5 to 20 wt. %, and more
preferably 5 to 8 wt. %, based on the total amount of the materials
of the equol-containing food.
[0239] Further, the proportion of equol in the equol-containing
food is generally 0.002 to 0.1 wt. %, preferably 0.004 to 0.05 wt.
%, and more preferably 0.005 to 0.03 wt. %, based on the total
amount of the materials of the equol-containing food. By ensuring
this proportion of the food material of the present invention, the
useful bioactivity derived from the equol-containing fermented
soybean hypocotyl is expressed in the food, while maintaining the
good flavor of the equol-containing food.
[0240] The equol-containing food is produced by mixing the food
material of the present invention with a predetermined amount of
the other ingredients, and subjecting the mixture to forming,
baking, cooling etc. depending on the target form of the food
product. The equol-containing food is made of an equol-containing
fermented soybean hypocotyl, and contains various useful
bioactivity materials including equol, thereby expressing various
bioactivities or pharmacological activities. Therefore, the
equol-containing food is useful not only as a general food but also
as a food for specified health use, nutritional supplement,
functional food, invalid food or the like. The food containing the
fermented soybean hypocotyl of the present invention is
particularly useful as a nutraceutical product.
[0241] For example, the equol-containing food has an effect of
preventing or treating various diseases or symptoms including
menopausal disorders, osteoporosis, prostatic hypertrophy, and
metabolic syndrome; or for blood cholesterol level reduction,
skin-whitening, pimples, intestinal control, obesity, and diuresis.
Particularly, the equol-containing food is suitable for prevention
or treatment of indefinite complaint or postmenopausal symptoms
(e.g., osteoporosis, menopausal disorder) of middle-aged women.
[0242] The intake of the equol-containing food per day is
determined depending on the equol content in the food, age, weight,
symptom of the consumer, or the dosage number.
4. Foods of Various Forms
[0243] The present invention further provides various forms of food
containing the equol-containing fermented soybean hypocotyl or
extract thereof. Examples of the foods of the present invention
includes beverages, dietary supplements, creamy foods, desserts,
confectionaries, seasonings, retort-packed foods, processed meat
products, fish pastes, processed egg products, canned foods,
breads, frozen desserts, processed soybean foods, cooked rice
products, and soup.
[0244] The following explains in detail the equol-containing
fermented soybean hypocotyl and the extract thereof, and each form
of food provided by the present invention.
Equol-Containing Fermented Soybean Hypocotyl and Extract
Thereof
[0245] The equol-containing fermented soybean hypocotyl or extract
thereof used for the present invention is the same as that used in
the section "3. Food Material".
[0246] The food of the present invention may contain any of the
items in the foregoing list of equol-containing fermented soybean
hypocotyls or extracts thereof. The food of the present invention
preferably contains an equol-containing fermented soybean
hypocotyl.
Forms of Food
[0247] The equol-containing fermented soybean hypocotyl or extract
thereof is effective in preventing or treating various diseases or
symptoms including menopausal disorders, osteoporosis, prostatic
hypertrophy, and metabolic syndrome; or for blood cholesterol level
reduction, skin-whitening, pimples, intestinal control, obesity,
and diuresis. Particularly, the equol-containing food is suitable
for the prevention or treatment of indefinite complaints or post
menopausal symptoms (e.g. osteoporosis, menopausal disorders) of
middle-aged women. Accordingly, the various forms of food of the
present invention may be provided as functional foods effective for
those diseases or symptoms. More specifically, the food of the
present invention containing fermented soybean hypocotyl is not
only useful as a general food, but also useful as a food for
specified health use, a nutritional supplement, a functional food,
an invalid diet food or the like. The food of the present invention
containing fermented soybean hypocotyl is particularly useful as a
nutritional supplement.
[0248] The following explains specific examples of the food forms
of the present invention.
(1) Beverage
[0249] The present invention provides a beverage containing the
equol-containing fermented soybean hypocotyl or extract
thereof.
[0250] The beverage of the present invention may be any kind of
beverage; however, it is suitable for beverages containing fruit
juice; beverages containing vegetable juice; carbonated beverages
such as sparkling water, cola, a fruit-juice-containing carbonated
beverage, ginger ale, soda; soft drinks such as a sports drink;
coffee beverages such as coffee or milk coffee; teas such as
English-style tea, green tea, or oolong tea; cocoa; lactic acid
bacteria beverages; milk beverages such as cow's milk,
fruit-flavored milk, or sour milk beverages; alcoholic beverages
such as beer, shochu (distilled spirit), whiskey, cocktails, or
shochu mixed with soda water.
[0251] The content of the equol-containing fermented soybean
hypocotyl or extract thereof in each beverage of the present
invention is not limited, and is individually determined according
to the equol content in the fermented soybean hypocotyl or extract
thereof, the type of beverage, the expected effect or the like. For
example, the content of the fermented soybean hypocotyl or extract
thereof in the beverage is generally 0.02 to 15 wt. %, preferably
0.1 to 1 wt. %, and more preferably 0.2 to 0.5 wt. % (dry
weight).
(2) Dietary Supplement
[0252] The present invention provides a dietary supplement
containing the equol-containing fermented soybean hypocotyl or
extract thereof.
[0253] The form of the dietary supplement of the present invention
is not limited, and includes a softgel, a tablet, powder or the
like, among which a softgel is particularly preferred. The softgel
or powder is preferably contained in a capsule at a predetermined
amount.
[0254] One of the suitable embodiments of the softgel of the
present invention is a softgel that contains gelatin, water and
glycerin having the equol-containing fermented soybean hypocotyl or
extract therein. The softgel may further contain other additives
such as soybean oil, beeswax, lecithin, or (3-carotin.
[0255] The content of the equol-containing fermented soybean
hypocotyl or extract thereof in each dietary supplement of the
present invention is not limited, and is individually determined
according to the equol content in the fermented soybean hypocotyl
or extract thereof, the type of supplement, the expected effect or
the like. For example, the content of the fermented soybean
hypocotyl or extract thereof in the dietary supplement is 2 to 100
wt. % (dry weight).
(3) Creamy Food
[0256] The present invention provides a creamy food containing the
equol-containing fermented soybean hypocotyl or extract
thereof.
[0257] The creamy food of the present invention is not particularly
limited and may include whipped cream, custard cream, butter cream,
fresh cream, and coffee whitener.
[0258] The content of the equol-containing fermented soybean
hypocotyl or extract thereof in each creamy food of the present
invention is not limited, and is individually determined according
to the equol content in the fermented soybean hypocotyl or extract
thereof, the type of creamy food, the expected effect or the like.
For example, the content of the fermented soybean hypocotyl or
extract thereof in the creamy food is generally 0.03 to 5 wt. %,
preferably 0.15 to 1 wt. %, and more preferably 0.3 to 0.5 wt. %
(dry weight).
(4) Dessert
[0259] The present invention provides a dessert containing the
equol-containing fermented soybean hypocotyl or extract
thereof.
[0260] The dessert is not particularly limited and may include
yoghurt, jelly, pudding, mousse or the like.
[0261] The content of the equol-containing fermented soybean
hypocotyl or extract thereof in each dessert of the present
invention is not limited, and is individually determined according
to the equol content in the fermented soybean hypocotyl or extract
thereof, the type of dessert, the expected effect or the like. For
example, the content of the fermented soybean hypocotyl or extract
thereof in the dessert is generally 0.03 to 5 wt. %, preferably
0.15 to 1 wt. %, and more preferably 0.3 to 0.5 wt. % (dry
weight).
(5) Snack
[0262] The present invention provides a snack containing the
equol-containing fermented soybean hypocotyl or extract
thereof.
[0263] The snack is not particularly limited and may include any
western or Japanese sweets including baked or steamed
confectionary. Examples of the snack of the present invention
include candies, lollies, caramels, pretzels, tablets, gums,
chocolates, cookies, bean confectionery, wafer, cheese crackers,
gummy candies, ordinary rice crackers, and Suhama (Japanese
traditional confectionery), dry confectionery, and Yokan (azuki
bean jelly), steamed bean-jam bun, bracken-starch dumplings, pound
cakes, apple pie fillings, savory snacks, and glutinous rice
crackers. Among them, one of the suitable embodiments of the snack
is a baked confectionery containing soy flour.
[0264] The content of the equol-containing fermented soybean
hypocotyl or extract thereof in each snack of the present invention
is not limited, and is individually determined according to the
equol content in the fermented soybean hypocotyl or extract
thereof, the type of snack, the expected effect or the like. For
example, the content of the fermented soybean hypocotyl or extract
thereof in the snack is generally 0.01 to 99 wt. %, preferably 0.04
to 36 wt. %, and more preferably 0.08 to 18 wt. % (dry weight).
(6) Seasoning
[0265] The present invention provides a seasoning containing the
equol-containing fermented soybean hypocotyl or extract
thereof.
[0266] The seasoning is not particularly limited and may include
barbecue sauce, sauce for Kabayaki (broiled seafood), seasoning of
squid-chinmi, ketchup, tsuyu (soup), dressing, sesame dipping
sauce, snack seasonings and soy sauce.
[0267] The content of the equol-containing fermented soybean
hypocotyl or extract thereof in each seasoning of the present
invention is not limited, and is individually determined according
to the equol content in the fermented soybean hypocotyl or extract
thereof, the type of seasoning, the expected effect or the like.
For example, the content of the fermented soybean hypocotyl or
extract thereof in the seasoning is generally 0.04 to 40 wt. %,
preferably 0.2 to 8 wt. %, and more preferably 0.4 to 4 wt. % (dry
weight).
(7) Retort-Packed Food
[0268] The present invention provides a retort-packed food (retort
pouch food) containing the equol-containing fermented soybean
hypocotyl or extract thereof.
[0269] The retort-packed food is not particularly limited and may
include various seasoning materials including a sauce premix for
curry sauce, hashed rice sauce, stew, meat sauce, or white sauce; a
premix for Chinese-style donburi, beef donburi, and chicken and egg
donburi; a premix for Ma-po Dou-fu (Tofu with chili and bean-based
sauce), and a premix for Kamameshi (a Japanese rice dish cooked
with various kinds of meat, seafood, and vegetables).
[0270] The content of the equol-containing fermented soybean
hypocotyl or extract thereof in each retort-packed food of the
present invention is not limited, and is individually determined
according to the equol content in the fermented soybean hypocotyl
or extract thereof, the type of retort-packed food, the expected
effect or the like. For example, the content of the fermented
soybean hypocotyl or extract thereof in the retort-packed food is
generally 0.003 to 4 wt. %, preferably 0.01 to 0.8 wt. %, and more
preferably 0.03 to 0.4 wt. % (dry weight).
(8) Processed Meat Product
[0271] The present invention provides a processed meat product
containing the equol-containing fermented soybean hypocotyl or
extract thereof.
[0272] The processed meat product of the present invention is not
particularly limited and may include sausages (dry, semi-dry,
bologna, wiener, smoke), liver paste, bacon, shoulder bacon, ham,
hamburger steak, and meatballs.
[0273] The content of the equol-containing fermented soybean
hypocotyl or extract thereof in each processed meat product of the
present invention is not limited, and is individually determined
according to the equol content in the fermented soybean hypocotyl
or extract thereof, the type of processed meat product, the
expected effect or the like. For example, the content of the
fermented soybean hypocotyl or extract thereof in the processed
meat product is generally 0.02 to 0.4 wt. %, preferably 0.1 to 0.8
wt. %, and more preferably 0.2 to 0.4 wt. % (dry weight).
(9) Fish Paste Product
[0274] The present invention provides a fish paste product
containing the equol-containing fermented soybean hypocotyl or
extract thereof.
[0275] The fish paste product of the present invention is not
particularly limited and may include boiled fish paste and fishcake
tubes.
[0276] The content of the equol-containing fermented soybean
hypocotyl or extract thereof in each fish paste product of the
present invention is not limited, and is individually determined
according to the equol content in the fermented soybean hypocotyl
or extract thereof, the type of fish paste product, the expected
effect or the like. For example, the content of the fermented
soybean hypocotyl or extract thereof in the fish paste product is
generally 0.03 to 5 wt. %, preferably 0.1 to 1 wt. %, and more
preferably 0.3 to 0.5 wt. % (dry weight).
(10) Processed Egg Product
[0277] The present invention provides a processed egg product
containing the equol-containing fermented soybean hypocotyl or
extract thereof.
[0278] The processed egg product of the present invention is not
particularly limited and may include Japanese rolled omelet, thick
Japanese omelet, western-style omelet, and scrambled eggs.
[0279] The content of the equol-containing fermented soybean
hypocotyl or extract thereof in each processed egg product of the
present invention is not limited, and is individually determined
according to the equol content in the fermented soybean hypocotyl
or extract thereof, the type of processed egg product, the expected
effect or the like. For example, the content of the fermented
soybean hypocotyl or extract thereof in the processed egg product
is generally 0.03 to 5 wt. %, preferably 0.1 to 1 wt. %, and more
preferably 0.3 to 0.5 wt. % (dry weight).
(11) Canned Food
[0280] The present invention provides a canned food containing the
equol-containing fermented soybean hypocotyl or extract
thereof.
[0281] The canned food of the present invention is not particularly
limited and may include canned fruit such as canned oranges or
canned peaches; canned seafood such as canned tuna or canned
sardines; canned jam; canned soup; and canned sauce.
[0282] The content of the equol-containing fermented soybean
hypocotyl or extract thereof in each canned food of the present
invention is not limited, and is individually determined according
to the equol content in the fermented soybean hypocotyl or extract
thereof, the type of canned food, the expected effect or the like.
For example, the content of the fermented soybean hypocotyl or
extract thereof in the canned food is generally 0.03 to 5 wt. %,
preferably 0.1 to 1 wt. %, and more preferably 0.3 to 0.5 wt. %
(dry weight).
(12) Bread
[0283] The present invention provides bread containing the
equol-containing fermented soybean hypocotyl or extract
thereof.
[0284] The bread of the present invention is not particularly
limited and may include a loaf of bread, French bread, stuffed
bread, melon bread, bean-jam bread, cream bread, jam bread, bread
rolls, and croissants.
[0285] The content of the equol-containing fermented soybean
hypocotyl or extract thereof in each bread of the present invention
is not limited, and is individually determined according to the
equol content in the fermented soybean hypocotyl or extract
thereof, the type of bread, the expected effect or the like. For
example, the content of the fermented soybean hypocotyl or extract
thereof in the bread is generally 0.01 to 50 wt. %, preferably 0.05
to 0.6 wt. %, and more preferably 0.1 to 0.3 wt. % (dry
weight).
(13) Frozen Dessert
[0286] The present invention provides a frozen dessert containing
the equol-containing fermented soybean hypocotyl or extract
thereof.
[0287] The bread of the present invention is not particularly
limited and may include ice cream and sherbet.
[0288] The content of the equol-containing fermented soybean
hypocotyl or extract thereof in each frozen dessert of the present
invention is not limited, and is individually determined according
to the equol content in the fermented soybean hypocotyl or extract
thereof, the type of frozen dessert, the expected effect or the
like. For example, the content of the fermented soybean hypocotyl
or extract thereof in the frozen dessert is generally 0.03 to 5 wt.
%, preferably 0.1 to 1 wt. %, and more preferably 0.3 to 0.5 wt. %
(dry weight).
(14) Processed Soybean Food
[0289] The present invention provides a processed soybean food
containing the equol-containing fermented soybean hypocotyl or
extract thereof.
[0290] The processed soybean food of the present invention is not
particularly limited and may include tofu, thick deep-fried tofu,
fried tofu mixed with vegetables, soy milk skin, fermented
soybeans, and soy milk.
[0291] The content of the equol-containing fermented soybean
hypocotyl or extract thereof in each processed soybean food of the
present invention is not limited, and is individually determined
according to the equol content in the fermented soybean hypocotyl
or extract thereof, the type of processed soybean food, the
expected effect or the like. For example, the content of the
fermented soybean hypocotyl or extract thereof in the processed
soybean food is generally 0.03 to 0.5 wt. %, preferably 0.1-1 wt.
%, and more preferably 0.3-0.5 wt. % (dry weight).
(15) Cooked Rice Product
[0292] The present invention provides a cooked rice product
containing the equol-containing fermented soybean hypocotyl or
extract thereof. The cooked rice product of the present invention
is not particularly limited and may include fried rice, pilaf, rice
gruel, steamed glutinous rice with chestnut, cooked white rice, red
beans rice, and rice steamed with meat and vegetables.
[0293] The content of the equol-containing fermented soybean
hypocotyl or extract thereof in each cooked rice product of the
present invention is not limited, and is individually determined
according to the equol content in the fermented soybean hypocotyl
or extract thereof, the type of cooked rice product, the expected
effect or the like. For example, the content of the fermented
soybean hypocotyl or extract thereof in the cooked rice product is
generally 0.01 to 2.5 wt. %, preferably 0.02 to 0.5 wt. %, and more
preferably 0.04 to 0.25 wt. % (dry weight).
(16) Soup
[0294] The present invention provides soup containing the
equol-containing fermented soybean hypocotyl or extract
thereof.
[0295] The soup of the present invention is not particularly
limited and may include corn cream soup, consomme, minestrone,
onion soup, and potage.
[0296] The content of the equol-containing fermented soybean
hypocotyl or extract thereof in each soup of the present invention
is not limited, and is individually determined according to the
equol content in the fermented soybean hypocotyl or extract
thereof, the type of soup, the expected effect or the like. For
example, the content of the fermented soybean hypocotyl or extract
thereof in the soup is generally 0.02 to 15 wt. %, preferably 0.1
to 1 wt. %, and more preferably 0.2 to 0.5 wt. % (dry weight).
EXAMPLES
[0297] The following more specifically explains the present
invention with reference to Reference Examples, Examples etc. The
present invention is however not limited to these examples.
Reference Examples 1-1 to 1-3
Production of Equol-Containing Fermented Soybean Hypocotyl
[0298] Soybean hypocotyl powder, arginine, and water were mixed to
prepare a soybean hypocotyl solution (material) having a
composition as shown in Table 1. A Lactococcus 20-92 strain (FERM
BP-10036) was inoculated in this soybean hypocotyl solution of 5
ml, and the sample was subjected to stationary culture for 96 hours
at 37.degree. C. under anaerobic conditions. After that, the
resulting fermentation solution (culture solution) was sterilized
by heating for a minute at 100.degree. C., followed by drying at
80.degree. C. The dried product was processed into a powder using a
homogenizer to obtain fermented soybean hypocotyl powder.
[0299] Table 1 shows the viable cell count and pH of the culture
after 96 hours culture, the amount of fermented soybean hypocotyl
powder collected and the equol content of the fermented soybean
hypocotyl powder. From the data, it is shown that fermentation of
soybean hypocotyl powder using equol-producing bacteria produces
equol with high efficiency.
TABLE-US-00001 TABLE 1 Ref.Ex. 1-1 Ref.Ex. 1-2 Ref.Ex. 1-3
Composition Soybean 0.25 g 0.5 g 0.75 g of Soybean Hypocotyl
Hypocotyl Powder Solution (dry wt.) (Material) Arginine 0.005 g
0.005 g 0.005 g Water Appropriate Appropriate Appropriate Quantity
Quantity Quantity Total 5 ml 5 ml 5 ml Amount PH 6.75 .+-. 0.03
6.54 .+-. 0.02 6.39 .+-. 0.03 Analytical Viable 7.9 .+-. 0.1 8.2
.+-. 0.1 8.3 .+-. 0.2 Result of Bacterial Fermented Counts of
Liquid Fermented Liquid (log cfu/ml) PH of 7.00 .+-. 0.03 6.88 .+-.
0.01 6.76 .+-. 0.02 Fermented Liquid Analysis of Equol 3.85 mg (--)
3.44 mg (--) 5.38 mg Composition (48.9 of wt. %) Fermented Daidzein
n. d. (--) n. d. 1.18 mg Soybean Compounds (10.7 wt. %) Hypocotyl
Genistein n. d. (--) n. d. 1.45 mg Powder in Compounds (13.2 terms
of wt. %) Isoflavone Glycitein n. d. (--) n. d. 3.00 mg Compounds
(27.2 wt. %)
[0300] Each Example was performed using three lots of soybean
hypocotyl powder (N=3). In the analysis of isoflavone composition
in the table, the value on the left denotes a content (mg) of each
isoflavone per gram of the fermented soybean hypocotyl, and the
value in the bracket on the right denotes the ratio (wt. %) of each
isoflavone to the gross isoflavone amount (100 wt. %) in the
fermented soybean hypocotyl. "n.d." indicates that the measurement
is not done, and "-" in the bracket indicates that the calculation
is not done.
Reference Example 1-4
Production of Equol-containing Fermented Soybean Hypocotyl
[0301] A Lactococcus 20-92 strain (FEM BP-10036) was inoculated to
a 5 ml soybean hypocotyl solution containing 10 wt % of soybean
hypocotyl powder and 0.1 wt. % of arginine, and the sample was
subjected to fermentation by carrying out stationary culture for 96
hours at 37.degree. C. under anaerobic conditions. After that, the
resulting fermentation solution (culture solution) was sterilized
by heating for a minute at 100.degree. C., followed by drying at
80.degree. C. The dried product was processed into powder using a
homogenizer to obtain fermented soybean hypocotyl powder.
[0302] The respective components of the soybean hypocotyl powder
("Pre-fermentation" in Table 2 and Table 3) and of the fermented
soybean hypocotyl powder ("Post-fermentation" in Table 2 and Table
3) were analyzed. Table 2 shows an analysis regarding soy
isoflavone, and Table 3 shows an analysis regarding nutritional
components. These tables show that the fermented soybean hypocotyl
obtained by the fermentation of soybean hypocotyl using a
Lactococcus 20-92 strain has a high equol content. Further,
oligosaccharide content for such oligosaccharides as raffinose or
stachyose remains substantially the same after fermentation, which
indicates that the fermentation did not influence the
oligosaccharide content. Meanwhile, the tables show that arginine
was converted into ornithine by the fermentation. Accordingly, by
adding arginine to the soybean hypocotyl before fermentation using
a Lactococcus 20-92 strain, the resulting fermented substance
contains not only equol but also ornithine.
TABLE-US-00002 TABLE 2 Soybean Isoflavones Per 100 g Component
Pre-fermentation Post-fermentation Equol N.D. 632.0 mg Daidzin
566.4 mg 29.7 mg Malonyldaidzin 124.9 mg N.D. Acetyldaidzin 364.8
mg 25.4 mg Daidzein 7.1 mg 24.4 mg Dihydrodaidzein N.D. 49.4 mg
Genistin 111.7 mg 3.2 mg Malonylgenistin 35.1 mg N.D.
Acetylgenistin 146.1 mg 3.7 mg Genistein 0.9 mg 22.5 mg
Dihydrogenistein N.D. 112.0 mg Glycitin 331.7 mg 53.6 mg
Malonylglycitin 65.0 mg N.D. Acetylglycitin 169.2 mg 34.8 mg
Glycitein 19.1 mg 292.3 mg Dihydroglycitein N.D. 8.2 mg Total
Isoflavones 1942.0 mg 1291.2 mg N.D. refers to "Not Detected"
TABLE-US-00003 TABLE 3 Nutritional Component Per 100 g Component
Pre-fermentation Post-fermentation Moisture 3.2 g 6.2 g Protein
38.1 g 38.3 g Fat 13.0 g 14.5 g Ash 4.3 g 4.0 g Saccharide 30.9 g
26.8 g Dietary Fiber 10.5 g 10.2 g Energy 414 kcal 411 kcal Sucrose
7.95 g 7.42 g Raffinose 1.37 g 1.34 g Stachyose 9.04 g 8.38 g Trans
Fatty Acids N.D. N.D. Phospholipids (as stear-, ol 3.33 g 2.92 g
e-, and lecithin) Free Arginine 881 mg 12 mg Free Ornithine N.D.
1.06 g Soyasapogenol A N.D. N.D. Soyasapogenol B N.D. N.D. Soybean
Saponin 3.6 g 3.8 g N.D. refers to "Not Detected"
Reference Examples 1-5 to 1-11
Production of Equol-Containing Fermented Soybean Hypocotyl
[0303] In these examples, fermented soybean hypocotyl powder
(Reference Examples 1-5 to 1-11) were produced in the same manner
as in Reference Examples 1-3 except that seven lots of soybean
hypocotyl powder different from the three lots of Reference Example
1-3 were used. The compositions of isoflavone in the obtained
fermented soybean hypocotyl was analyzed as shown in Table 4. This
result also shows that each of the fermented soybean hypocotyls of
Reference Examples 1-5 to 1-11 has a high equol content with
isoflavone proportion that was not attained by any known arts.
TABLE-US-00004 TABLE 4 Isoflavone Proportions Daidzein Genistein
Glycitein Equol Compounds Compounds Compounds Ref.Ex. 6.51 mg 0.71
mg 0.53 mg 2.71 mg 1-5 (62.2 wt. %) (6.8 wt. %) (5.1 wt. %) (25.9
wt. %) Ref.Ex. 6.25 mg 0.48 mg 0.35 mg 3.12 mg 1-6 (61.3 wt. %)
(4.7 wt. %) (3.4 wt. %) (30.6 wt. %) Ref.Ex. 5.38 mg 1.18 mg 1.45
mg 3.00 mg 1-7 (48.9 wt. %) (10.7 wt. %) (13.2 wt. %) (27.2 wt. %)
Ref.Ex. 6.43 mg 0.61 mg 0.48 mg 2.62 mg 1-8 (63.4 wt. %) (6.0 wt.
%) (4.7 wt. %) (25.8 wt. %) Ref.Ex. 6.05 mg 0.51 mg 0.30 mg 2.57 mg
1-9 (64.2 wt. %) (5.4 wt. %) (3.2 wt. %) (27.3 wt. %) Ref.Ex. 6.11
mg 0.37 mg 0.10 mg 2.74 mg 1-10 (65.6 wt. %) (4.0 wt. %) (1.1 wt.
%) (29.4 wt. %) Ref.Ex. 6.3 mg 0.49 mg 0.37 mg 3.19 mg 1-11 (60.9
wt. %) (4.73 wt. %) (3.6 wt. %) (30.8 wt. %)
[0304] The value in the upper column denotes a content (mg) of each
isoflavone per gram of the fermented soybean hypocotyl, and the
value in the lower column denotes the ratio (wt. %) of each
isoflavone to the gross isoflavone amount (100 wt. %) in the
fermented soybean hypocotyl.
Reference Experiment Example 1
Allergen Confirmation Test
[0305] Soybean hypocotyl is known to contain allergens including
Gym4, Gm30K, Gm28K, 7S globulin mix (3-conglycine), oleosin, and
trypsin inhibitors. The existence of allergens in the
equol-containing fermented soybean hypocotyl produced in Reference
Example 1-1 was examined using the following test.
[0306] First, an appropriate amount of the equol-containing
fermented soybean hypocotyl obtained in Reference Example 1-1 was
added to an extraction buffer (Tris HCl, pH 7.5, containing 1M EDTA
and an appropriate amount of protease inhibitor). The sample was
stirred well to extract a water-soluble component. The resulting
solution was filtered to remove the solid content to obtain a
liquid extract. All proteins contained in this liquid extract were
detected using a protein assay system (Bio-Rad). Further, the major
allergens (Gym4, Gm30K, Gm28K, 7S globulin mix, oleosin, and
trypsin inhibitor) contained in the liquid extract were detected
using western blotting. Further, for comparison, the detection of
total protein and allergen was carried out in the same manner using
soybean cotyledon powder and soybean hypocotyl powder, instead of
the fermented soybean hypocotyl.
[0307] FIGS. 1 to 3 show the result. FIG. 1 shows the result for
the detection of total protein; FIG. 2 shows the result for the
detection of Gym4, Gm30K, and Gm28K; and FIG. 3 shows the result
for the detection of 7S globulin mix, oleosin, and trypsin
inhibitor.
[0308] These results show that the major allergen content of a
soybean or a soybean hypocotyl is reduced in the equol-containing
fermented soybean hypocotyl.
Example 1: Production of Equol-Containing Extract
Example 1-1: Production of Equol-containing Extract by Production
Method I
[0309] One gram of the equol-containing fermented soybean hypocotyl
powder produced under the same conditions as in Reference Examples
1 to 3 was added to 20 ml of the extractant according to Table 5;
and 120 minutes of shaking was performed at room temperature.
Thereafter, the residue ("Residue 1", hereinafter) was removed by
centrifugation to isolate a liquid extract ("Liquid Extract 1-1",
hereinafter). Next, Liquid Extract 1-1 was evaporated to dryness
using an evaporator, 5 ml of ethanol was added thereto, and 10
minutes of shaking was performed at room temperature. Thereafter,
the residue ("Residue 2", hereinafter) was removed by
centrifugation to isolate a liquid extract ("Liquid Extract 1-2",
hereinafter). Liquid Extract 1-2 was then evaporated to dryness
using an evaporator to obtain a solid extract.
[0310] Further, another 20 ml of extractant of the same composition
was added to Residue 1, and 120 minutes shaking was performed at
room temperature. Thereafter, the residue was removed by
centrifugation to isolate a liquid extract (Liquid Extract 2-1,
hereinafter). Liquid Extract 2-1 was then evaporated to dryness
using an evaporator. 5 ml of ethanol was added thereto, and 10
minutes of shaking was performed at room temperature. Thereafter,
the residue was removed by centrifugation to isolate an extract
("Liquid Extract 2-2", hereinafter). Liquid Extract 2-2 was
evaporated to dryness using an evaporator to obtain a solid
extract.
TABLE-US-00005 TABLE 5 Name of Extractant Composition of Extractant
25Et Ethanol Solution containing 25 vol. % of Ethanol and 75 vol. %
of Purified Water 50Et Ethanol Solution containing 50 vol. % of
Ethanol and 50 vol. % of Purified Water 75Et Ethanol Solution
containing 75 vol. % of Ethanol and 25 vol. % of Purified Water
100Et 100 vol. % of Ethanol
[0311] Liquid Extracts 1-1 and 1-2, Liquid Extracts 2-1 and 2-2,
and Residue 2 were subjected to thin-layer chromatography (TLC)
analysis (as a mobile phase, hexane:chloroform:ethyl acetate=1:1:1
(volume ratio) was used for equol detection; and ethyl
acetate:propanol:chloroform:methanol:0.9% KCl=25:25:25:10:9 (volume
ratio) was used for saponin detection) so as to measure saponin
content and equol content. The weight of the solid extract obtained
from Liquid Extract 1-2 and its equol concentration were measured.
FIGS. 4 to 7 and Table 6 show the results. FIG. 4 shows the results
of TLC analysis of the equol contents in Liquid Extracts 1-2 and
2-2; FIG. 5 shows the results of TLC analysis of the saponin
contents in Liquid Extracts 1-2 and 2-2; FIG. 6 shows the results
of TLC analysis of the equol contents in Liquid Extract 1-2 and
Residue 2; and FIG. 7 shows the results of TLC analysis of the
saponin contents in Liquid Extract 1-2 and Residue 2.
[0312] According to the figures, it was found that when the
equol-containing fermented soybean hypocotyl was subjected to the
extraction using ethanol, the extract contained a large amount of
oil, and the equol recovery rate was low.
[0313] In contrast, when the equol-containing fermented soybean
hypocotyl was subjected to the extraction using an aqueous ethanol,
the inclusion of oil was suppressed, and the equol recovery rate
increased. Particularly, it became clear that use of an aqueous
ethanol solution having an ethanol concentration of 75 vol. %
significantly increases the equol collection rate. Further, it was
also found that extraction using a 75 vol. % ethanol minimizes the
extraction of fat components. Furthermore, according to the fact
that saponin in a concentrated amount was detected in Residue 2,
saponin can be removed without reducing the equol amount by
subjecting the residue, which was obtained by removing solvent from
Liquid Extract 1-1, to extraction using ethanol.
TABLE-US-00006 TABLE 6 EQL-containing Fermented Soybean Solid
Extract Collected from Hypocotyl Liquid Extract 1-2 Amount EQL EQL
EQL Used for EQL Concen- Dry EQL Concen- EQL Concen- Extract
Extraction Content tration Weight Content tration Collection
tration Type (g) (mg) (wt. %) (mg) (mg) (wt. %) Rate (%) Rate 25Et
1 5.38 0.54 43 1.78 4.14 33.1 7.7 50Et 1 5.38 0.54 36 2.68 7.44
49.8 13.8 75Et 1 5.38 0.54 121 5.56 4.60 103.4 8.5 100Et 1 5.38
0.54 146 1.50 1.03 27.9 1.9
[0314] EQL represents equol. The "concentration rate" represents
the EQL concentration rate of the solid extract when assuming the
EQL concentration in the EQL-containing fermented soybean hypocotyl
is 1.
Example 1-2
Production of Equol-Containing Extract by Production Method I
[0315] In this example, equol-containing fermented soybean
hypocotyl powder was produced under the same conditions as in
Reference Example 1-3 except that a different lot of soybean
hypocotyl powder was used.
[0316] 30 g of the equol-containing fermented soybean hypocotyl
powder was added to 150 ml of a 70 vol. % aqueous ethanol solution
(100 ml consisting of ethanol and 30 ml of water) or 150 ml of a 90
vol. % aqueous ethanol solution (100 ml consisting of ethanol and
10 ml of water); and eight hours stirring was performed at room
temperature. After removing the residue by filtration, the residue
was washed with a 20 ml of aqueous ethanol solution having the same
composition as that of the extractant. The filtrate and the liquid
after washing were collected to obtain a liquid extract. The liquid
extract was concentrated under reduced pressure to obtain a solid
extract.
[0317] The weight of the obtained solid extract and the weight of
equol contained therein and the concentrations of daidzein,
dihydrodaidzein, genistein, dihydrogenistein, glystein,
dihydroglystein were measured, as shown in Tables 7 and 8.
TABLE-US-00007 TABLE 7 Extraction Results for 70 vol. % Ethanol
Solution EQL-containing Fermented Soybean Hypocotyl Collected Solid
Extract Amount for Dry Weight Weight Extraction (g) 30 (mg) 8000
EQL EQL EQL EQL EQL EQL EQL Content Concentration Content
Concentration Collection Concentration (mg) (wt. %) (mg) (wt. %)
Rate (%) Rate (%) 161.40 0.54 158.98 1.98 98.5 3.7 DZN DZN DZN DZN
DZN DZN DZN Content Concentration Content Concentration Collection
Concentration (mg) (wt. %) (mg) (wt. %) Rate (%) Rate (%) 6.00 0.02
0.00 0.00 0.0 0.0 DHD DHD DHD DHD DHD DHD DHD Content Concentration
Content Concentration Collection Concentration (mg) (wt. %) (mg)
(wt. %) Rate (%) Rate (%) 9.00 0.03 0.09 0.00 0.00 0.00 GNT GNT GNT
GNT GNT GNT GNT Content Concentration Content Concentration
Collection Concentration (mg) (wt. %) (mg) (wt. %) Rate (%) Rate
(%) 6.90 0.02 0.31 0.00 4.5 0.2 DHG DHG DHG DHG DHG DHG DHG Content
Concentration Content Concentration Collection Concentration (mg)
(wt. %) (mg) (wt. %) Rate (%) Rate (%) 32.7 0.11 6.96 0.09 21.3 0.5
GLY GLY GLY GLY GLY GLY GLY Content Concentration Content
Concentration Collection Concentration (mg) (wt. %) (mg) (wt. %)
Rate (%) Rate (%) 62.70 0.21 33.17 0.41 52.9 2.0 DHG DHG DHG DHG
DHG DHG DHG Content Concentration Content Concentration Collection
Concentration (mg) (wt. %) (mg) (wt. %) Rate (%) Rate (%) 0.9 0.00
0.00 0.00 0.00 0.00
[0318] EQL represents equol; DZN represents daidzein; DHD
represents dihydrodaidzein; GNT represents genistein; DHG
represents dihydrogenistein; GLY represents glystein; and DHG
represents dihydroglystein.
[0319] The isoflavone content (mg) denotes the gross weight of each
isoflavone contained in the EQL-containing fermented soybean
hypocotyl (material: 30 g) or in the obtained isolated extract
(8000 mg); and the isoflavone concentration (wt. %) denotes the
proportion of each isoflavone contained in the EQL-containing
fermented soybean hypocotyl (material) or in the obtained isolated
extract. The "Concentration Rate" represents the concentration rate
of each component in the solid extract when assuming the
concentration of the component in the EQL-containing fermented
soybean hypocotyl is 1.
TABLE-US-00008 TABLE 8 Extraction Results for 90 vol. % Ethanol
Solution EQL-containing Fermented Soybean Hypocotyl Collected Solid
Extract Amount for Dry Weight Weight Extraction (g) 30 (mg) 4100
EQL EQL EQL EQL EQL EQL EQL Content Concentration Content
Concentration Collection Concentration (mg) (wt. %) (mg) (wt. %)
Rate (%) Rate (%) 161.40 0.54 158.13 3.89 98.0 7.2 DZN DZN DZN DZN
DZN DZN DZN Content Concentration Content Concentration Collection
Concentration (mg) (wt. %) (mg) (wt. %) Rate (%) Rate (%) 6.00 0.02
0.00 0.00 0.0 0.0 DHD DHD DHD DHD DHD DHD DHD Content Concentration
Content Concentration Collection Concentration (mg) (wt. %) (mg)
(wt. %) Rate (%) Rate (%) 9.00 0.03 0.12 0.00 1.3 0.1 GNT GNT GNT
GNT GNT GNT GNT Content Concentration Content Concentration
Collection Concentration (mg) (wt. %) (mg) (wt. %) Rate (%) Rate
(%) 6.90 0.02 0.34 0.01 4.9 0.4 DHG DHG DHG DHG DHG DHG DHG Content
Concentration Content Concentration Collection Concentration (mg)
(wt. %) (mg) (wt. %) Rate (%) Rate (%) 32.7 0.11 6.89 0.17 21.1 1.4
GLY GLY GLY GLY GLY GLY GLY Content Concentration Content
Concentration Collection Concentration (mg) (wt. %) (mg) (wt. %)
Rate (%) Rate (%) 62.70 0.21 32.64 0.80 52.1 3.8 DHG DHG DHG DHG
DHG DHG DHG Content Concentration Content Concentration Collection
Concentration (mg) (wt. %) (mg) (wt. %) Rate (%) Rate (%) 0.9 0.00
0.00 0.00 0.00 0.00
[0320] EQL represents equol; DZN represents daidzein; DHD
represents dihydrodaidzein; GNT represents genistein; DHG
represents dihydrogenistein; GLY represents glystein; and DHG
represents dihydroglystein.
[0321] The isoflavone content (mg) denotes the gross weight of each
isoflavone contained in the EQL-containing fermented soybean
hypocotyl (material: 30 g) or in the obtained isolated extract
(4100 mg); and the isoflavone concentration (wt. %) denotes the
proportion of each isoflavone contained in the EQL-containing
fermented soybean hypocotyl (material) or in the obtained isolated
extract. The "Concentration Rate" represents the concentration rate
of each component in the solid extract when assuming the
concentration of the component in the EQL-containing fermented
soybean hypocotyl is 1.
[0322] The tables show that, by carrying out extraction on an
equol-containing fermented soybean hypocotyl using an aqueous
ethanol solution, the equol collection rate increases and glystein
was also collected. Meanwhile, the concentrations of other
isoflavones were all very low in this extract.
Example 1-3
Production of Equol-Containing Extract by Production Method I
[0323] In this example, equol-containing fermented soybean
hypocotyl powder was produced under the same conditions as in
Reference Example 1-3 except that a different lot of soybean
hypocotyl powder was used.
[0324] 50 g of the equol-containing fermented soybean hypocotyl
powder was mixed with 7 ml of purified water, and then further
mixed with 250 ml of n-hexane; and seven hours of stirring was
performed at room temperature. Insoluble residue ware isolated by
filtration, and seven hours of drying was performed at 60.degree.
C. to obtain defatted equol-containing fermented soybean hypocotyl
powder.
[0325] 20 g of the defatted equol-containing fermented soybean
hypocotyl powder was added to 100 ml of a 80 vol. % aqueous ethanol
solution (100 ml consisting of ethanol and 20 ml of water) or 100
ml of a 90 vol. % aqueous ethanol solution (100 ml consisting of
ethanol and 10 ml of water), and fourteen hours of stirring was
performed at room temperature. After removing the residue by
filtration, the residue was washed using 10 ml of an aqueous
ethanol solution having the same composition as that of the
extractant. The filtrate and the liquid after washing were isolated
to obtain a liquid extract.
[0326] The liquid extract was dried for six hours at 60.degree. C.
to obtain a solid extract.
[0327] The weight of the obtained solid extract and the weight of
equol contained therein and the concentrations of daidzein,
dihydrodaidzein, genistein, dihydrogenistein, glystein,
dihydroglystein were measured, as shown in Tables 9 and 10.
TABLE-US-00009 TABLE 9 Extraction Results for 80 vol. % Ethanol
Solution Defatted EQL-containing Fermented Soybean Hypocotyl powder
Collected Solid Extract Amount for Dry Weight Weight Extraction (g)
20 (mg) 4000 EQL EQL EQL EQL EQL EQL EQL Content Concentration
Content Concentration Collection Concentration (mg) (wt. %) (mg)
(wt. %) Rate (%) Rate (%) 107.60 0.54 114.89 2.88 106.8 5.3 DZN DZN
DZN DZN DZN DZN DZN Content Concentration Content Concentration
Collection Concentration (mg) (wt. %) (mg) (wt. %) Rate (%) Rate
(%) 4.00 0.02 0.00 0.00 0.0 0.0 DHD DHD DHD DHD DHD DHD DHD Content
Concentration Content Concentration Collection Concentration (mg)
(wt. %) (mg) (wt. %) Rate (%) Rate (%) 6.00 0.03 0.10 0.00 1.7 0.1
GNT GNT GNT GNT GNT GNT GNT Content Concentration Content
Concentration Collection Concentration (mg) (wt. %) (mg) (wt. %)
Rate (%) Rate (%) 4.60 0.02 0.28 0.01 0.01 0.4 DHG DHG DHG DHG DHG
DHG DHG Content Concentration Content Concentration Collection
Concentration (mg) (wt. %) (mg) (wt. %) Rate (%) Rate (%) 32.7 0.11
6.96 0.13 21.3 0.9 GLY GLY GLY GLY GLY GLY GLY Content
Concentration Content Concentration Collection Concentration (mg)
(wt. %) (mg) (wt. %) Rate (%) Rate (%) 41.80 0.21 24.03 0.60 57.5
2.9 DHG DHG DHG DHG DHG DHG DHG Content Concentration Content
Concentration Collection Concentration (mg) (wt. %) (mg) (wt. %)
Rate (%) Rate (%) 0.6 0.00 0.00 0.00 0.00 0.00
[0328] EQL represents equol; DZN represents daidzein; DHD
represents dihydrodaidzein; GNT represents genistein; DHG
represents dihydrogenistein; GLY represents glystein; and DHG
represents dihydroglystein.
[0329] The isoflavone content (mg) denotes the total weight of each
isoflavone contained in the defatted EQL-containing fermented
soybean hypocotyl (material: 20 g) or in the collected extract
(4000 mg); and the isoflavone concentration (wt. %) denotes the
proportion of each isoflavone contained in the defatted
EQL-containing fermented soybean hypocotyl (material) or in the
collected extract. The "Concentration Rate" represents the
concentration rate of each component in the solid extract when
assuming the concentration of the component in the defatted
EQL-containing fermented soybean hypocotyl is 1.
TABLE-US-00010 TABLE 10 Extraction Results for 90 vol. % Ethanol
Solution Defatted EQL-containing Fermented Soybean Hypocotyl powder
Collected Solid Extract Amount for Dry Weight Weight Extraction (g)
20 (mg) 2200 EQL EQL EQL EQL EQL EQL EQL Content Concentration
Content Concentration Collection Concentration (mg) (wt. %) (mg)
(wt. %) Rate (%) Rate (%) 107.60 0.54 106.88 4.81 99.3 8.9 DZN DZN
DZN DZN DZN DZN DZN Content Concentration Content Concentration
Collection Concentration (mg) (wt. %) (mg) (wt. %) Rate (%) Rate
(%) 4.00 0.02 0.00 0.00 0.0 0.0 DHD DHD DHD DHD DHD DHD DHD Content
Concentration Content Concentration Collection Concentration (mg)
(wt. %) (mg) (wt. %) Rate (%) Rate (%) 6.00 0.03 0.10 0.00 1.7 0.2
GNT GNT GNT GNT GNT GNT GNT Content Concentration Content
Concentration Collection Concentration (mg) (wt. %) (mg) (wt. %)
Rate (%) Rate (%) 4.60 0.02 0.21 0.01 0.01 0.4 DHG DHG DHG DHG DHG
DHG DHG Content Concentration Content Concentration Collection
Concentration (mg) (wt. %) (mg) (wt. %) Rate (%) Rate (%) 32.7 0.11
6.96 0.21 21.3 1.5 GLY GLY GLY GLY GLY GLY GLY Content
Concentration Content Concentration Collection Concentration (mg)
(wt. %) (mg) (wt. %) Rate (%) Rate (%) 41.80 0.21 21.97 0.99 52.6
4.7 DHG DHG DHG DHG DHG DHG DHG Content Concentration Content
Concentration Collection Concentration (mg) (wt. %) (mg) (wt. %)
Rate (%) Rate (%) 0.6 0.00 0.00 0.00 0.00 0.00
[0330] EQL represents equol; DZN represents daidzein; DHD
represents dihydrodaidzein; GNT represents genistein; DHG
represents dihydrogenistein; GLY represents glystein; and DHG
represents dihydroglystein.
[0331] The isoflavone content (mg) denotes the gross weight of each
isoflavone contained in the defatted EQL-containing fermented
soybean hypocotyl (material: 20 g) or in the collected extract
(2200 mg); and the isoflavone concentration (wt. %) denotes the
proportion of each isoflavone contained in the defatted
EQL-containing fermented soybean hypocotyl (material) or in the
collected extract. The "Concentration Rate" represents the
concentration rate of each component in the solid extract when
assuming the concentration of the component in the defatted
EQL-containing fermented soybean hypocotyl is 1.
[0332] The tables show that the high equol collection rate obtained
in Example 1-2 was ensured even when defatting the equol-containing
fermented soybean hypocotyl in advance.
Example 1-4
Production of Equol-containing Extract by Production Method II
[0333] In this example, equol-containing fermented soybean
hypocotyl powder (equol concentration=6.2 mg/g) was produced under
the same conditions as in Reference Example 1-3 except that a
different lot of soybean hypocotyl powder was used.
[0334] 80 g of the equol-containing fermented soybean hypocotyl
powder was suspended in 900 ml of water. 5 ml of acetic acid was
added to the suspension while it was being stirred, until the pH
became approximately 4.0. After 24 hours standing at room
temperature, insoluble residue was collected by filtration, and
dried. 50.33 g of insoluble residue was thus obtained.
[0335] Then, 250 ml of an aqueous ethanol solution (ethanol
concentration=75 vol. %) was added to the 50.33 g of insoluble
residue, and two days stirring was performed at room temperature.
After removing the residue by filtration under reduced pressure,
the filtrate was collected. The filtrate was evaporated to dryness
into a solid with an evaporator to obtain 7.53 g of
equol-containing powder.
[0336] The equol concentration of the obtained equol-containing
powder was 60.0 mg/g, which was about ten times the equol
concentration (6.2 mg/g) of the equol-containing fermented soybean
hypocotyl which was used as the starting material. The recovery
rate of equol was 91%.
Example 2: Equol Purification
Example 2-1: Equol Purification Method 1
[0337] Equol-containing fermented soybean hypocotyl powder (equol
concentration=6.2 mg/g) was produced under the same conditions as
in Reference Example 1-3 except that a different lot of soybean
hypocotyl powder was used.
[0338] 10 g of the equol-containing fermented soybean hypocotyl
powder was added to 50 ml of a 50 vol. % aqueous ethanol solution,
and two hours shaking was performed at room temperature.
Thereafter, the residue ("Residue 1", hereinafter) was removed by
centrifugation (2500 rpm, 25 minutes) to isolate a liquid extract
("Liquid Extract A-1", hereinafter). Next, 50 ml of a 50 vol. %
aqueous ethanol solution was added to Residue 1, and an hour of
shaking was performed at room temperature. Thereafter, the residue
was removed by centrifugation (2500 rpm, 25 minutes) to isolate a
liquid extract ("Liquid Extract A-2", hereinafter). Extracts A-1
and A-2 were mixed and then evaporated to dryness using an
evaporator to obtain a residual solid ("Residual Solid 1",
hereinafter).
[0339] Then, 10 ml of ethanol was added to Residual Solid 1, and 30
minutes of shaking was performed at room temperature under
supersonic treatment. Thereafter, the residue was removed by
centrifugation (1000 rpm, 10 minutes) to isolate a liquid extract
("Liquid Extract B", hereinafter). Liquid Extract B was evaporated
to dryness using an evaporator to obtain about 1 g of a residual
solid ("Residual Solid 2", hereinafter).
[0340] Next, 10 ml of hexane was added to the 1 g of Residual Solid
2, and 30 minutes of shaking was performed at room temperature.
Then, the liquid extract was removed by centrifugation (1000 rpm,
10 minutes) to collect 0.3 g of insoluble residue.
[0341] 2 ml of a mixed solution of hexane and dimethylether
(hexane:dimethylether=2:3 (volume ratio)) was added to the obtained
0.3 mg of insoluble residue, and 30 minutes shaking was performed
at room temperature. Then, the residue was removed by
centrifugation (1000 rpm, 10 minutes) to collect a liquid extract.
The obtained liquid extract was subjected to silica gel
chromatography under the following conditions.
[0342] Column: A column 2 cm in internal diameter, filled with
silica gel.
[0343] Mobile Phase: [0344] 200 ml of a mixed solution of hexane
and diethylether (hexane:diethylether=40:60 (volume ratio)); [0345]
200 ml of a mixed solution of hexane and diethylether
(hexane:diethylether=30:70 (volume ratio)); [0346] 200 ml of a
mixed solution of hexane and diethylether
(hexane:diethylether=25:75 (volume ratio)); [0347] 200 ml of a
mixed solution of hexane and diethylether
(hexane:diethylether=20:80 (volume ratio)); [0348] and 200 ml of
hexane were supplied sequentially.
[0349] Every 50 ml elution from the column was collected. The
fraction eluted by the mixed solution of hexane and diethylether
(hexane:diethylether=40:60 (volume ratio)) and the mixed solution
of hexane and diethylether (hexane:diethylether=25:75 (volume
ratio)) contained 40 mg of equol (recovery rate=80%), and the
purity of equol was 99.9% per dry solid. FIG. 8 shows the TLC
Analysis of the fraction eluted using silica gel chromatography. As
shown in the figure, the fraction eluted by the mixed solution of
hexane and diethylether (hexane:diethylether=30:70 (volume ratio))
and the mixed solution of hexane and diethylether
(hexane:diethylether=25:75 (volume ratio)) contained equol with
high purity. Table 11 details the samples supplied to the lanes of
FIG. 8.
TABLE-US-00011 TABLE 11 Lane in FIG. 8 Sample Details Lane 1
Standard Product of Equol Lane 2 Elution Fraction obtained while
the elution amount of Mixed Solution of Hexane and Diethylether
(Hexane:Diethylether = 40:60 (Capacity Ratio)) ranges between 0 to
50 ml Lane 3 Elution Fraction obtained while the elution amount of
Mixed Solution of Hexane and Diethylether (Hexane:Diethylether =
40:60 (Capacity Ratio)) ranges between 50 to 100 ml Lane 4 Elution
Fraction obtained while the elution amount of Mixed Solution of
Hexane and Diethylether (Hexane:Diethylether = 30:70 (Capacity
Ratio)) ranges between 0 to 50 ml Lane 5 Elution Fraction obtained
while the elution amount of Mixed Solution of Hexane and
Diethylether (Hexane:Diethylether = 30:70 (Capacity Ratio)) ranges
between 50 to 100 ml Lane 6 Elution Fraction obtained while the
elution amount of Mixed Solution of Hexane and Diethylether
(Hexane:Diethylether = 25:75 (Capacity Ratio)) ranges between 0 to
50 ml Lane 7 Elution Fraction obtained while the elution amount of
Mixed Solution of Hexane and Diethylether (Hexane:Diethylether =
25:75 (Capacity Ratio))ranges between 50 to 100 ml Lane 8 Elution
Fraction obtained while the elution amount of Mixed Solution of
Hexane and Diethylether (Hexane:Diethylether = 20:80 (Capacity
Ratio)) ranges between 0 to 50 ml
Example 2-2: Equol Purification Method 2
[0350] Equol-containing fermented soybean hypocotyl powder (equol
concentration=6.2 mg/g) was produced under the same conditions as
in Reference Example 1-3 except that a different lot of soybean
hypocotyl powder was used.
[0351] 100 g of the equol-containing fermented soybean hypocotyl
powder was added to 500 ml of a 95 vol. % aqueous ethyl acetate
solution, and four hours of stirring was performed at room
temperature. Thereafter, insoluble residue were removed by
filtration to collect an extract. The separated insoluble residue
was washed with 80 ml of ethyl acetate, and the liquid after
washing was mixed with the filtrate, and the mixture was
concentrated under reduced pressure to collect 12 g of oily matter.
The oily matter was subjected to silica gel chromatography.
[0352] Column: A column 2 cm in internal diameter, filled with 70 g
of silica gel using hexane.
[0353] Mobile phase: Supplied with 500 ml of hexane, and a mixed
solution of ethyl acetate and hexane (ethyl acetate:hexane=1:5
(volume ratio)) subsequently.
[0354] The solvent was distilled off from the equol-containing
elution solvent thus collected, and 630 mg of a yellow crystalline
solid was obtained.
[0355] The crystalline solid was added to 2 ml of ethyl acetate and
dissolved by heating at 80.degree. C. 8 ml of hexane was added
thereto and 30 minutes of stirring was performed while cooling the
mixture with ice to precipitate equol crystals. The obtained equol
crystals were collected by filtration, and washed with hexane. Then
the crystals were dried to obtain 530 mg of equol-containing solid
(pale yellow crystals).
[0356] Further, the 530 mg equol-containing solid was added to 4 ml
of aqueous ethanol solution (ethanol concentration=62.5 vol. %),
and heated to 80.degree. C. to be dissolved in the solution. The
solution was allowed to stand for twelve hours at 10.degree. C. to
recrystallize equol, and then filtered and dried. As a result, 390
mg of white needle-shaped equol crystals were obtained.
[0357] The angle of rotation [.alpha.].sub.D.sup.25 of the obtained
equol crystals was -21.6.degree. in a 25.degree. C. ethanol. The
fusing point was 188-190.degree. C. It was confirmed that the
obtained equol was nearly 100% pure.
Example 3: Food Material Containing an Equol-Containing Fermented
Soybean Hypocotyl or Extract Thereof, and Foods Containing the Food
Material
Example 3-1 Production of Food Material Containing an
Equol-Containing Fermented Soybean Hypocotyl and Cacao Mass
[0358] An equol-containing fermented soybean hypocotyl produced
under the same conditions as in Reference Examples 1-3 was mixed
with cacao mass having been dissolved by heat at a ratio of 1:3
(weight ratio). The mixture was cooled to obtain a food material in
which an equol-containing fermented soybean hypocotyl is dispersed
in cacao mass.
[0359] The taste of the food material thus produced was evaluated
by 10 panelists, with the result that all of the panelists
concluded that the bitter taste derived from the equol-containing
fermented soybean hypocotyl was suppressed, and the food material
possessed a pleasing flavor.
Example 3-2 Food Material Containing an Extract of Equol-Containing
Fermented Soybean Hypocotyl
[0360] A solid extract ("75Et extract", hereinafter) obtained by
evaporating Liquid Extract 2-1 produced in Example 1-1 using 75Et
was mixed with cacao mass having been dissolved by heat at a ratio
of 1:24 (weight ratio). The mixture was cooled to obtain a food
material in which a 75Et extract was dispersed in cacao mass.
[0361] The taste of the food material thus produced was evaluated
by 10 panelists, with the result that all of the panelists
concluded that the bitter taste derived from the 75Et extract was
suppressed, and the food material possessed a pleasing flavor.
Example 3-3 Baked Confectionery
[0362] The ingredients shown in Table 12 were mixed, and the
mixture was formed into a rectangle (about 1.5 cm.times.1.5
cm.times.10 cm) and baked in an oven at 180.degree. C. to produce a
baked confectionery.
[0363] The taste of the baked confectionery thus produced was
evaluated by 10 panelists, with the result that all of the
panelists concluded that the bitter taste derived from the
equol-containing fermented soybean hypocotyl was suppressed, and
the baked confectionery possessed a pleasing chocolate flavor and
an original pleasing taste.
TABLE-US-00012 TABLE 12 Proportion (wt. %) Chip-shaped Food
Material 5 Obtained in Example 3-1 (about 0.1 g per chip) Soy Flour
25 Raisins q.s. Butter q.s. Sugar q.s. Egg q.s. Orange Peel q.s.
Pineapple q.s. Indigestible Dextrin q.s. Cacao Mass q.s. Sliced
Almonds q.s. Cocoa Powder q.s. Salt q.s. Baking Powder q.s. Flavor
q.s. Total 100
Comparative Example 3-1: Baked Confectionery
[0364] A baked confectionery was produced under the same conditions
as in Example 3-3 except that 1 g of the equol-containing fermented
soybean hypocotyl produced under the same conditions as in Example
1-3 was mixed with 5 g of chocolate chips instead of the
chip-shaped food material obtained in Example 3-1.
[0365] The taste of the baked confectionery thus produced was
evaluated by 10 panelists, with the result that all of the
panelists concluded that the pleasing taste disappeared and the
bitter taste derived from the equol-containing fermented soybean
hypocotyl remained throughout the confectionary.
Example 3-4: Baked Confectionery
[0366] A baked confectionery was produced under the same conditions
as in Example 3-3 except for the inclusion of the same amount of
chip-shaped food material obtained in Example 3-2 instead of the
chip-shaped food materials obtained in Example 3-1.
[0367] The taste of the baked confectionery thus produced was
evaluated by 10 panelists, with the result that all of the
panelists concluded that the bitter taste derived from the 75Et
extract was suppressed, and the baked confectionery possessed a
pleasing chocolate flavor and an original pleasing taste.
Example 4 Food Comprising Equol-Containing Fermented Soybean
Hypocotyl Material or its Extract
Example 4-A Beverage
[0368] Hereinafter, specific production examples of the beverage
according to the present invention are given.
Example 4-A-1 Sparkling Water
[0369] Sparkling water was obtained by preparing a mint-flavored
syrup having the following composition, and mixing the resultant
mint-flavored syrup with carbonated water in a volumetric ratio of
3:1.
TABLE-US-00013 Composition of Mint-flavored Syrup Equol-containing
fermented soybean 0.27 (Kg) hypocotyl material (Reference Example
1-3) Sugar 0.60 Citric acid (crystalline) 0.01 Table Salt 0.012
Potassium chloride 0.005 Calcium lactate 0.008 Peppermint extract
0.03 Peppermint flavor 0.20 Water Balance Total 100.00 L
Example 4-A-2 Sparkling Water
[0370] Sparkling water was produced under the same conditions as in
Example 4-A-1, except that the mint-flavored syrup was prepared by
adding, instead of the equol-containing fermented soybean hypocotyl
material, 0.034 kg of the solid extract prepared in Example 1-1 by
evaporating Liquid Extract 1-2 obtained using 75Et to dryness.
Example 4-A-3 Orange-flavored Carbonated Beverage Containing No
Orange Juice
[0371] An orange-flavored carbonated beverage containing no orange
juice was obtained by preparing an orange-flavored syrup having the
following composition, and mixing the resultant orange-flavored
syrup with carbonated water in a volumetric ratio of 9:11.
TABLE-US-00014 Composition of Orange-flavored Syrup
Equol-containing fermented soybean 0.20 (Kg) hypocotyl material
(Reference-Example 1-3) Sugar 10.00 Citric acid (crystalline) 0.03
DL-malic acid 0.07 Food coloring 0.05 Flavor 0.10 Water Balance
Total 45.00 L
Example 4-A-4 Orange-flavored Carbonated Beverage Containing No
Orange Juice
[0372] An orange-flavored carbonated beverage containing no orange
juice was produced under the same conditions as in Example 4-A-3,
except that the orange-flavored syrup was prepared by adding,
instead of the equol-containing fermented soybean hypocotyl
material, 0.025 kg of the solid extract prepared in Example 1-1 by
evaporating Liquid Extract 1-2 obtained using 75Et to dryness.
Example 4-A-5 Low-Calorie Cola
[0373] Sugarless cola was obtained by preparing a cola syrup having
the following composition, and mixing the resultant cola syrup with
carbonated water in a volumetric ratio of 8:12.
TABLE-US-00015 Composition of Cola Syrup Equol-containing fermented
soybean hypocotyl 0.20 (Kg) material (Reference Example 1-2)
Sucralose 0.02 Phosphoric acid 0.05 Citric acid (crystalline) 0.07
Caffeine 0.10 Caramel coloring 0.20 Flavor (cola flavor) 0.10 Water
Balance Total 40.00 L
Example 4-A-6 Low-Calorie Cola
[0374] Low-calorie cola was produced under the same conditions as
in Example 4-A-5, except that the cola syrup was prepared by
adding, instead of the equol-containing fermented soybean hypocotyl
material, 0.03 kg of the solid extract prepared in Example 1-1 by
evaporating Liquid Extract 1-2 obtained using 75Et to dryness.
Example 4-A-7 Carbonated Apple Beverage
[0375] A carbonated apple beverage was obtained by preparing
apple-flavored syrup having the following composition, and mixing
the resultant apple-flavored syrup with carbonated water in a
volumetric ratio of 13:7.
TABLE-US-00016 Composition of Apple-flavored Syrup Equol-containing
fermented soybean hypocotyl 0.20 (Kg) material (Reference Example
1-2) 5 times concentrated apple juice 11.00 Fructose/glucose syrup
5.00 Citric acid (crystalline) 0.10 Food coloring 0.05 Flavor 0.10
Water Balance Total 65.00 L
Example 4-A-8 Carbonated Apple Beverage
[0376] A carbonated apple beverage was produced under the same
conditions as in Example 4-A-7, except that the apple-flavored
syrup was prepared by adding, instead of the equol-containing
fermented soybean hypocotyl material, 0.03 kg of the solid extract
prepared in Example 1-1 by evaporating Liquid Extract 1-2 obtained
using 75Et to dryness.
Example 4-A-9 Carbonated Grape Beverage
[0377] A carbonated grape beverage was obtained by preparing a
grape-flavored syrup having the following composition, and mixing
the resultant grape-flavored syrup with carbonated water in a
volumetric ratio of 9:11.
TABLE-US-00017 Composition of Grape-flavored Syrup Equol-containing
fermented soybean hypocotyl 0.20 (Kg) material (Reference Example
1-1) 5 times concentrated clear grape juice 4.40 Citric acid
(crystalline) 0.25 Food coloring 0.05 Flavor 0.20 Water Balance
Total 45.00 L
Example 4-A-10 Carbonated Grape Beverage
[0378] A carbonated grape beverage was produced under the same
conditions as in Example 4-A-9, except that the grape-flavored
syrup was prepared by adding, instead of the equol-containing
fermented soybean hypocotyl material, 0.03 kg of the solid extract
prepared in Example 1-1 by evaporating Liquid Extract 1-2 obtained
using 75Et to dryness.
Example 4-A-11 Beverage Containing Apple Juice
[0379] A beverage containing apple juice having the following
composition was produced.
TABLE-US-00018 Equol-containing fermented soybean hypocotyl 0.40
(Kg) material (Reference Example 1-3) 5 times concentrated apple
juice 22.00 Flavor 0.05 Water Balance Total 100.00 L
Example 4-A-12 Beverage Containing Apple Juice
[0380] A beverage containing apple juice was produced under the
same conditions as in Example 4-A-11, except for using, instead of
the equol-containing fermented soybean hypocotyl material, 0.05 kg
of the solid extract prepared in Example 1-1 by evaporating Liquid
Extract 1-2 obtained using 75Et to dryness.
Example 4-A-13 Beverage Containing Orange Juice
[0381] A beverage containing orange juice having the following
composition was produced.
TABLE-US-00019 Equol-containing fermented soybean hypocotyl 0.40
(Kg) material (Reference Example 1-3) Concentrated Valencia orange
juice 4.40 (Brix: 55 degree) Sugar 5.00 Citric acid (crystalline)
0.16 Native gellan gum 0.025 Vitamin C 0.03 Pectin 0.0025 Orange
flavor 0.25 Water Balance Total 100.00 L
Example 4-A-14 Beverage Containing Orange Juice
[0382] A beverage containing orange juice was produced under the
same conditions as in Example 4-A-13, except for using, instead of
the equol-containing fermented soybean hypocotyl material, 0.05 kg
of the solid extract prepared in Example 1-1 by evaporating Liquid
Extract 1-2 obtained using 75Et to dryness.
Example 4-A-15 Low-calorie Beverage Containing Fruit Juice
[0383] A low-calorie beverage containing fruit juice having the
following composition was produced.
TABLE-US-00020 Equol-containing fermented soybean hypocotyl 0.40
(Kg) material (Reference Example 1-1) 5 times concentrated citrus
juice 4.40 Sucralose 0.009 Citric acid (crystalline) 0.18
L-ascorbic acid 0.03 Gellan gum 0.024 Flavor 0.10 Water Balance
Total 100.00 L
Example 4-A-16 Low-Calorie Beverage Containing Fruit Juice
[0384] A low-calorie beverage containing fruit juice was produced
under the same conditions as in Example 4-A-15, except for using,
instead of the equol-containing fermented soybean hypocotyl
material, 0.05 kg of the solid extract prepared in Example 1-1 by
evaporating Liquid Extract 1-2 obtained using 75Et to dryness.
Example 4-A-17 Sugarless Near Water (Soft Drink Similar to
Water)
[0385] Sugarless near water (soft drink similar to water) having
the following composition was produced.
TABLE-US-00021 Equol-containing fermented soybean hypocotyl 0.40
(Kg) material (Reference Example 1-3) Sucralose 0.008 Polydextrose
2.20 5 times concentrated grapefruit juice 0.44 Citric acid
(crystalline) 0.53 Calcium lactate 0.06 Potassium chloride 0.01
L-ascorbic acid 0.03 Dibenzoyl thiamine hydrochloride 0.0002
Pyridoxine hydrochloride 0.00015 Nicotinamide 0.0015 Flavor 0.10
Water Balance Total 100.00 L
Example 4-A-18 Sugarless Near Water (Soft Drink Similar to
Water)
[0386] Sugarless near water (soft drink similar to water) was
produced under the same conditions as in Example 4-A-17, except for
using, instead of the equol-containing fermented soybean hypocotyl
material, 0.05 kg of the solid extract prepared in Example 1-1 by
evaporating Liquid Extract 1-2 obtained using 75Et to dryness.
Example 4-A-19 Sports Drink
[0387] A sports drink having the following composition was
produced.
TABLE-US-00022 Equol-containing fermented soybean hypocotyl 0.40
(Kg) material (Reference Example 1-2) Clear lemon juice 0.50
Sucralose 0.02 Lactic minerals 1.00 Vitamin mix 0.25 Sodium
L-ascorbate 0.05 Citric acid 0.125 Water Balance Total 100.00 L
Example 4-A-20 Sports Drink
[0388] A sports drink was produced under the same conditions as in
Example 4-A-19, except for using, instead of the equol-containing
fermented soybean hypocotyl material, 0.05 kg of the solid extract
prepared in Example 1-1 by evaporating Liquid Extract 1-2 obtained
using 75Et to dryness.
Example 4-A-21 Powdered Green Tea-Milk Drink
[0389] A powdered green tea-milk drink having the following
composition was produced.
TABLE-US-00023 Equol-containing fermented soybean 0.40 (Kg)
hypocotyl material (Reference Example 1-2) Cow's milk 10.00 Skimmed
milk 3.50 Powdered green tea 0.50 (RS-20, produced by ITO EN, LTD.)
Sugar 7.00 Emulsifier 0.70 Food coloring 0.03 Flavor 0.20 Sodium
hydrogen carbonate for adjusting pH to 6.8 Water Balance Total
100.00 L
Example 4-A-22 Powdered Green Tea-Milk Drink
[0390] A powdered green tea-milk drink was produced under the same
conditions as in Example 4-A-21, except for using, instead of the
equol-containing fermented soybean hypocotyl material, 0.05 kg of
the solid extract prepared in Example 1-1 by evaporating Liquid
Extract 1-2 obtained using 75Et to dryness.
Example 4-A-23 Sugarless Yogurt Drink
[0391] A sugarless yogurt drink having the following composition
was produced.
TABLE-US-00024 Equol-containing fermented soybean hypocotyl 0.40
(Kg) material (Reference Example 1-3) Fermented milk (SNF20%) 40.00
Water-soluble soybean polysaccharide 0.20 Pectin 0.20 Flavor 0.20
Lactic acid for adjusting pH to 4.2 Water Balance Total 100.00
L
Example 4-A-24 Sugarless Yogurt Drink
[0392] A sugarless yogurt drink was produced under the same
conditions as in Example 4-A-23, except for using, instead of the
equol-containing fermented soybean hypocotyl material, 0.05 kg of
the solid extract prepared in Example 1-1 by evaporating Liquid
Extract 1-2 obtained using 75Et to dryness.
Example 4-A-25 Pasteurized Lactic Acid Bacteria Beverage
[0393] A pasteurized lactic acid bacteria beverage having the
following composition was produced.
TABLE-US-00025 Equol-containing fermented soybean hypocotyl 0.40
(Kg) material (Reference Example 1-3) Fermented milk (SNF20%) 15.00
Water-soluble soybean polysaccharide 0.40 Sugar 8.00 Flavor 0.10
Lactic acid for adjusting pH to 3.8 Water Balance Total 100.00
L
Example 4-A-26 Pasteurized Lactic Acid Bacteria Beverage
[0394] A pasteurized lactic acid bacteria beverage was produced
under the same conditions as in Example 4-A-25, except for using,
instead of the equol-containing fermented soybean hypocotyl
material, 0.05 kg of the solid extract prepared in Example 1-1 by
evaporating Liquid Extract 1-2 obtained using 75Et to dryness.
Example 4-A-27 Low-Calorie Sour Milk Beverage
[0395] A low-calorie sour milk beverage having the following
composition was produced.
TABLE-US-00026 Equol-containing fermented soybean hypocotyl 0.40
(Kg) material (Reference Example 1-3) Skimmed milk 3.50
Water-soluble soybean polysaccharide 0.40 Sugar 5.00 Sucralose
0.0006 Fructose/glucose syrup (75%) 5.00 Citric acid (crystalline)
0.45 Flavor 0.10 Lactic acid for adjusting pH to 3.8 Water Balance
Total 100.00 L
Example 4-A-28 Low-Calorie Sour Milk Beverage
[0396] Low-calorie sour milk beverage was produced under the same
conditions as in Example 4-A-27, except for using, instead of the
equol-containing fermented soybean hypocotyl material, 0.05 kg of
the solid extract prepared in Example 1-1 by evaporating Liquid
Extract 1-2 obtained using 75Et to dryness.
Example 4-A-29 Strawberry-Flavored Milk Drink
[0397] A strawberry-flavored milk drink having the following
composition was produced.
TABLE-US-00027 Equol-containing fermented soybean hypocotyl 0.40
(Kg) material (Reference Example 1-3) Skimmed milk 2.80 Whole milk
powder 3.00 Sugar 12.00 Emulsifier 0.06 Food coloring 0.05 Flavor
(strawberry flavor) 0.10 Water Balance Total 100.00 L
Example 4-A-30 Strawberry-Flavored Milk Drink
[0398] Strawberry-flavored milk drink was produced under the same
conditions as in Example 4-A-29, except for using, instead of the
equol-containing fermented soybean hypocotyl material, 0.05 kg of
the solid extract prepared in Example 1-1 by evaporating Liquid
Extract 1-2 obtained using 75Et to dryness.
Example 4-A-31 Non-Fat Milk
[0399] Non-fat milk having the following composition was
produced.
TABLE-US-00028 Equol-containing fermented soybean hypocotyl 0.40
(Kg) material (Reference Example 1-1) Skimmed milk 12.00 Egg
calcium 1.00 Emulsifier 0.06 Flavor 0.10 Water Balance Total 100.00
L
Example 4-A-32 Non-Fat Milk
[0400] Non-fat milk was produced under the same conditions as in
Example 4-A-31, except for using, instead of the equol-containing
fermented soybean hypocotyl material, 0.05 kg of the solid extract
prepared in Example 1-1 by evaporating Liquid Extract 1-2 obtained
using 75Et to dryness.
Example 4-A-33 Cream Soda
[0401] A cream soda having the following composition was
produced.
TABLE-US-00029 Equol-containing fermented soybean hypocotyl 0.20
(Kg) material (Reference Example 1-1) Skimmed sweetened condensed
milk 3.00 Citric acid (crystalline) 0.04 Sugar 15.00 Food coloring
0.60 Flavor 0.10 Water Balance Total 40.00 L
Example 4-A-34 Cream Soda
[0402] A cream soda was produced under the same conditions as in
Example 4-A-33, except for using, instead of the equol-containing
fermented soybean hypocotyl material, 0.025 kg of the solid extract
prepared in Example 1-1 by evaporating Liquid Extract 1-2 obtained
using 75Et to dryness.
Example 4-A-35 Powdered Green Tea-Milk Drink
[0403] A powdered green tea-milk drink having the following
composition was produced.
TABLE-US-00030 Equol-containing fermented soybean hypocotyl 0.40
(Kg) material (Reference Example 1-2) Skimmed milk 3.50 Cow's milk
10.00 Emulsifier 0.70 Sugar 6.00 Water Balance Total 100.00 L
Example 4-A-36 Powdered Green Tea-milk Drink
[0404] Powdered green tea-milk drink was produced under the same
conditions as in Example 4-A-35, except for using, instead of the
equol-containing fermented soybean hypocotyl material, 0.05 kg of
the solid extract prepared in Example 1-1 by evaporating Liquid
Extract 1-2 obtained using 75Et to dryness.
Example 4-A-37 Black Tea
[0405] Black tea having the following composition was produced.
TABLE-US-00031 Equol-containing fermented soybean hypocotyl 0.40
(Kg) material (Reference Example 1-2) Darjeeling tea extract (40
times) 20.00 Sugar 5.00 Flavor 0.05 Water Balance Total 100.00
L
Example 4-A-38 Black Tea
[0406] Black tea was produced under the same conditions as in
Example 4-A-37, except for using, instead of the equol-containing
fermented soybean hypocotyl material, 0.05 kg of the solid extract
prepared in Example 1-1 by evaporating Liquid Extract 1-2 obtained
using 75Et to dryness.
Example 4-A-39 Beverage Containing Apple Juice
[0407] A beverage containing apple juice having the following
composition was produced.
TABLE-US-00032 Equol-containing fermented soybean hypocotyl 0.40
(Kg) material (Reference Example 1-3) 5 times concentrated clear
apple juice 2.20 Oligosaccharide 1.00 Reducing maltose syrup 1.50
Vitamin C 0.05 Citric acid 0.10 Water-soluble dietary fiber 8.00
Water Balance Total 100.00 L
Example 4-A-40 Beverage Containing Apple Juice
[0408] A beverage containing apple juice was produced under the
same conditions as in Example 4-A-39, except for using, instead of
the equol-containing fermented soybean hypocotyl material, 0.05 kg
of the solid extract prepared in Example 1-1 by evaporating Liquid
Extract 1-2 obtained using 75Et to dryness.
Example 4-A-41 Soft Drink
[0409] A soft drink having the following composition was
produced.
TABLE-US-00033 Equol-containing fermented soybean hypocotyl 0.40
(Kg) material (Reference Example 1-3) Fructose 3.70
Fructose/glucose syrup 8.00 Sugar 0.30 DL-malic acid 0.07 Sodium
citrate 0.03 Emulsifier 0.052 Flavor 0.22 Water Balance Total
100.00 L
Example 4-A-42 Soft Drink
[0410] A soft drink was produced under the same conditions as in
Example 4-A-41, except for using, instead of the equol-containing
fermented soybean hypocotyl material, 0.05 kg of the solid extract
prepared in Example 1-1 by evaporating Liquid Extract 1-2 obtained
using 75Et to dryness.
Example 4-A-43 Beverage Containing Orange Juice
[0411] A beverage containing orange juice having the following
composition was produced.
TABLE-US-00034 Equol-containing fermented soybean hypocotyl 0.40
(Kg) material (Reference Example 1-3) Fructose/glucose syrup 2.50
Beet oligosaccharide 0.026 Sugar 4.00 Concentrated orange juice
4.40 Citric acid 0.026 Sodium citrate 0.003 L-ascorbic acid 0.01
Flavor 0.10 Water Balance Total 100.00 L
Example 4-A-44 Beverage Containing Orange Juice
[0412] A beverage containing orange juice was produced under the
same conditions as in Example 4-A-43, except for using, instead of
the equol-containing fermented soybean hypocotyl material, 0.05 kg
of the solid extract prepared in Example 1-1 by evaporating Liquid
Extract 1-2 obtained using 75Et to dryness.
Example 4-A-45 Beverage Containing Apple Juice
[0413] A beverage containing apple juice having the following
composition was produced.
TABLE-US-00035 Equol-containing fermented soybean hypocotyl 0.40
(Kg) material (Reference Example 1-3) Concentrated apple juice 4.40
Sugar 8.00 Citric acid 0.15 Flavor 0.12 Water Balance Total 100.00
L
Example 4-A-46 Beverage Containing Apple Juice
[0414] A beverage containing apple juice was produced under the
same conditions as in Example 4-A-45, except for using, instead of
the equol-containing fermented soybean hypocotyl material, 0.05 kg
of the solid extract prepared in Example 1-1 by evaporating Liquid
Extract 1-2 obtained using 75Et to dryness.
Example 4-A-47-1 Vegetable Juice
[0415] Vegetable juice having the following composition was
produced.
TABLE-US-00036 Equol-containing fermented soybean hypocotyl 0.40
(Kg) material (Reference Example 1-2) Vegetable fruit juice 20.00
Water Balance Total 100.00 L
Example 4-A-47-2 Vegetable Juice
[0416] Vegetable juice was produced under the same conditions as in
Example 4-A-47-1, except for using, instead of the equol-containing
fermented soybean hypocotyl material, 0.05 kg of the solid extract
prepared in Example 1-1 by evaporating Liquid Extract 1-2 obtained
using 75Et to dryness.
Example 4-A-48-1 Sports Drink
[0417] A sports drink having the following composition was
produced.
TABLE-US-00037 Equol-containing fermented soybean hypocotyl 0.40
(Kg) material (Reference Example 1-2) Lactic minerals 1.00 Vitamin
mix 0.25 Citric acid 0.125 Sodium L-ascorbate 0.05 Lemon juice 0.50
Sugar 12.00 Water Balance Total 100.00 L
Example 4-A-48-2 Sports Drink
[0418] A sports drink was produced under the same conditions as in
Example 4-A-48-1, except for using, instead of the equol-containing
fermented soybean hypocotyl material, 0.05 kg of the solid extract
prepared in Example 1-1 by evaporating Liquid Extract 1-2 obtained
using 75Et to dryness.
Example 4-A-49 Oolong Tea
[0419] Oolong tea having the following composition was
produced.
TABLE-US-00038 Equol-containing fermented soybean hypocotyl 0.40
(Kg) material (Reference Example 1-3) Oolong tea extract 3.00
Sodium L-ascorbate 0.01 Flavor 0.20 Water Balance Total 100.00
L
Example 4-A-50 Oolong Tea
[0420] Oolong tea was produced under the same conditions as in
Example 4-A-49, except for using, instead of the equol-containing
fermented soybean hypocotyl material, 0.05 kg of the solid extract
prepared in Example 1-1 by evaporating Liquid Extract 1-2 obtained
using 75Et to dryness.
Example 4-A-51 Tea with Lemon
[0421] Tea with lemon having the following composition was
produced.
TABLE-US-00039 Equol-containing fermented soybean hypocotyl 0.40
(Kg) material (Reference Example 1-3) Black tea extract 6.00
Reducing maltose syrup powder 3.00 Sugar 3.00 Lemon juice 0.50
L-ascorbic acid 0.02 Flavor 0.10 Water Balance Total 100.00 L
Example 4-A-52 Tea with Lemon
[0422] Tea with lemon was produced under the same conditions as in
Example 4-A-51, except for using, instead of the equol-containing
fermented soybean hypocotyl material, 0.05 kg of the solid extract
prepared in Example 1-1 by evaporating Liquid Extract 1-2 obtained
using 75Et to dryness.
Example 4-A-53 Tea with Milk
[0423] Tea with milk having the following composition was
produced.
TABLE-US-00040 Equol-containing fermented soybean hypocotyl 0.40
(Kg) material (Reference Example 1-3) Cow's milk 7.00 Whole milk
powder 0.25 Sugar 4.00 Tea extract 35.00 Emulsifier 0.05 Flavor
0.10 Water Balance Total 100.00 L
Example 4-A-54 Tea with Milk
[0424] Tea with milk was produced under the same conditions as in
Example 4-A-53, except for using, instead of the equol-containing
fermented soybean hypocotyl material, 0.05 kg of the solid extract
prepared in Example 1-1 by evaporating Liquid Extract 1-2 obtained
using 75Et to dryness.
Example 4-A-55 Coffee Drink
[0425] A coffee drink having the following composition was
produced.
TABLE-US-00041 Equol-containing fermented soybean hypocotyl 0.40
(Kg) material (Reference Example 1-1) Coffee extract 30.00 Cow's
milk 25.00 Emulsifier 0.10 Sodium bicarbonate 0.12 Stevia 0.01
Sugar 3.00 L-rhamnose 0.01 Flavor 0.05 Water Balance Total 100.00
L
Example 4-A-56 Coffee Drink
[0426] A coffee drink was produced under the same conditions as in
Example 4-A-55, except for using, instead of the equol-containing
fermented soybean hypocotyl material, 0.05 kg of the solid extract
prepared in Example 1-1 by evaporating Liquid Extract 1-2 obtained
using 75Et to dryness.
Example 4-A-57 Cocoa Milk Drink
[0427] A Cocoa milk drink having the following composition was
produced.
TABLE-US-00042 Equol-containing fermented soybean hypocotyl 0.40
(Kg) material (Reference Example 1-1) Cow's milk 10.00 Skimmed milk
3.50 Fructose 5.50 Sugar 0.40 Cocoa powder 1.00 Emulsifier 0.45
Flavor 0.08 Water Balance Total 100.00 L
Example 4-A-58 Cocoa Milk Drink
[0428] A cocoa milk drink was produced under the same conditions as
in Example 4-A-57, except for using, instead of the
equol-containing fermented soybean hypocotyl material, 0.05 kg of
the solid extract prepared in Example 1-1 by evaporating Liquid
Extract 1-2 obtained using 75Et to dryness.
Example 4-A-59 Powdered Green Tea-Milk Drink
[0429] A powdered green tea-milk drink having the following
composition was produced.
TABLE-US-00043 Equol-containing fermented soybean hypocotyl 0.40
(Kg) material (Reference Example 1-1) Cow's milk 10.00 Skimmed milk
3.50 Fructose 5.80 Sugar 0.40 Powdered green tea 0.90 Emulsifier
0.48 Flavor 0.27 Food coloring 0.03 Water Balance Total 100.00
L
Example 4-A-60 Powdered Green Tea-Milk Drink
[0430] A powdered green tea-milk drink was produced under the same
conditions as in Example 4-A-59, except for using, instead of the
equol-containing fermented soybean hypocotyl material, 0.05 kg of
the solid extract prepared in Example 1-1 by evaporating Liquid
Extract 1-2 obtained using 75Et to dryness.
Example 4-A-61 Sour Milk Beverage
[0431] A sour milk beverage having the following composition was
produced.
TABLE-US-00044 Equol-containing fermented soybean hypocotyl 0.40
(Kg) material (Reference Example 1-3) Skimmed milk 1.50 Fructose
2.50 Sugar 0.50 Concentrated strawberry juice 0.22 Sodium citrate
0.32 Emulsifier 0.40 Flavor 0.12 Water Balance Total 100.00 L
Example 4-A-62 Sour Milk Beverage
[0432] A sour milk beverage was produced under the same conditions
as in Example 4-A-61, except for using, instead of the
equol-containing fermented soybean hypocotyl material, 0.05 kg of
the solid extract prepared in Example 1-1 by evaporating Liquid
Extract 1-2 obtained using 75Et to dryness.
Example 4-A-63 Beverage Containing Vitamins
[0433] A beverage containing vitamins having the following
composition was produced.
TABLE-US-00045 Equol-containing fermented soybean hypocotyl
material (Reference Example 1-3) 0.40 (Kg) Sugar 24.00 Citric acid
0.20 L-ascorbic acid 0.30 Vitamin mix 0.06 Flavor 0.10 Carbonated
water Balance Total 100.00 L
Example 4-A-64 Beverage Containing Vitamins
[0434] A beverage containing vitamins was produced under the same
conditions as in Example 4-A-63, except for using, instead of the
equol-containing fermented soybean hypocotyl material, 0.05 kg of
the solid extract prepared in Example 1-1 by evaporating Liquid
Extract 1-2 obtained using 75Et to dryness.
Example 4-A-65 Red Bean Soup
[0435] A red bean soup of the following composition was
produced.
TABLE-US-00046 Equol-containing fermented soybean hypocotyl
material (Reference Example 1-3) 0.40 (Kg) Red bean paste 10.00
Sugar alcohol 6.50 Sugar 24.00 Table salt 0.10 Emulsifier 0.60
Flavor 0.10 Water Balance Total 100.00 L
Example 4-A-66 Red Bean Soup
[0436] A red bean soup was produced under the same conditions as in
Example 4-A-65, except for using, instead of the equol-containing
fermented soybean hypocotyl material, 0.05 kg of the solid extract
prepared in Example 1-1 by evaporating Liquid Extract 1-2 obtained
using 75Et to dryness.
Example 4-A-67 Green Tea
[0437] Green tea having the following composition was produced.
TABLE-US-00047 Equol-containing fermented soybean hypocotyl
material (Reference Example 1-1) 0.40 (Kg) Green tea extract 3.00
Sodium L-ascorbate 0.01 Flavor 0.20 Water Balance Total 100.00
L
Example 4-A-68 Green Tea
[0438] Green tea was produced under the same conditions as in
Example 4-A-67, except for using, instead of the equol-containing
fermented soybean hypocotyl material, 0.05 kg of the solid extract
prepared in Example 1-1 by evaporating Liquid Extract 1-2 obtained
using 75Et to dryness.
Example 4-A-69 Powdered Orange Juice Drink
[0439] A powdered orange juice drink having the following
composition was produced.
TABLE-US-00048 Equol-containing fermented soybean hypocotyl
material (Reference Example 1-2) 0.40 (Kg) Fructose/glucose syrup
2.50 Beet oligosaccharide 0.026 Sugar 4.00 Powdered orange juice
2.00 Citric acid 0.026 Sodium citrate 0.003 L-ascorbic acid 0.01
Flavor 0.10 Water Balance Total 100.00 L
Example 4-A-70 Powdered Orange Juice Drink
[0440] A powdered orange juice drink was produced under the same
conditions as in Example 4-A-69, except for using, instead of the
equol-containing fermented soybean hypocotyl material, 0.05 kg of
the solid extract prepared in Example 1-1 by evaporating Liquid
Extract 1-2 obtained using 75Et to dryness.
Example 4-A-71 Amino Acid Drink
[0441] An amino acid drink having the following composition was
produced.
TABLE-US-00049 Equol-containing fermented soybean hypocotyl
material (Reference Example 1-3) 0.40 (Kg) Sugar 15.00 Fructose
15.00 Citric acid 4.00 Sodium citrate 1.00 L-valine 2.00 L-leucine
4.00 L-isoleucine 2.00 L-arginine 2.00 Flavor 0.10 Water Balance
Total 100.00 L
Example 4-A-72 Amino Acid Drink
[0442] An amino acid drink was produced under the same conditions
as in Example 4-A-71, except for using, instead of the
equol-containing fermented soybean hypocotyl material, 0.05 kg of
the solid extract prepared in Example 1-1 by evaporating Liquid
Extract 1-2 obtained using 75Et to dryness.
Example 4-A-73 Soya Milk Drink
[0443] A soya milk drink having the following composition was
produced.
TABLE-US-00050 Equol-containing fermented soybean hypocotyl
material (Reference Example 1-3) 0.40 (part by weight) Soya Milk
100.00
Example 4-A-74 Soya Milk Drink
[0444] A soya milk drink was produced under the same conditions as
in Example 4-A-73, except for using, instead of the
equol-containing fermented soybean hypocotyl material, 0.05 parts
by weight of the solid extract prepared in Example 1-1 by
evaporating Liquid Extract 1-2 obtained using 75Et to dryness.
Example 4-A-75 Energy Drink
[0445] An energy drink having the following composition was
produced.
TABLE-US-00051 Equol-containing fermented soybean hypocotyl
material (Reference Example 1-3) 0.40 (Kg) Taurine 1.00
Nicotinamide 0.02 Vitamin B1 0.005 Vitamin B2 0.005 Vitamin B6
0.005 Caffeine 0.05 Sugar 15.00 Fructose 15.00 Citric acid 0.10
Flavor 0.10 Water Balance Total 100.00 L
Example 4-A-76 Amino Acid Drink
[0446] An amino acid drink was produced under the same conditions
as in Example 4-A-75, except for using, instead of the
equol-containing fermented soybean hypocotyl material, 0.05 kg of
the solid extract prepared in Example 1-1 by evaporating Liquid
Extract 1-2 obtained using 75Et to dryness.
Example 4-B Dietary Supplement
[0447] Hereinafter, specific production examples of the dietary
supplement according to the present invention are given.
Example 4-B-1 Softgel
[0448] A softgel was produced according to a known method using
equol-containing fermented soybean hypocotyl material (Reference
Example 1-3), gelatin, glycerine, citric acid, soybean oil,
beeswax, lecithin, .beta. carotene, and water.
Example 4-B-2 Softgel
[0449] A softgel was produced under the same conditions as in
Example 4-B-1, except for using, instead of the equol-containing
fermented soybean hypocotyl material, the solid extract prepared in
Example 1-1 by evaporating Liquid Extract 1-2 obtained using 75Et
to dryness.
Example 4-C Creamy Food
[0450] Hereinafter, specific production examples of the creamy food
according to the present invention are given.
Example 4-C-1 Whipped Cream
[0451] Whipped cream having the following composition was
produced.
TABLE-US-00052 Equol-containing fermented soybean hypocotyl
material (Reference Example 1-3) 0.40 (g) Coconut oil (mp:
35.degree. C.) 27.00 Glycerine fatty acid ester 0.10 Skimmed milk
3.30 Sucralose 0.012 Reducing starch sugar 25.00 Lecithin 0.15
Emulsifier 0.70 Sodium metaphosphate 0.10 Water Balance Total
100.00 g
Example 4-C-2 Whipped Cream
[0452] Whipped cream was produced under the same conditions as in
Example 4-C-1, except for using, instead of the equol-containing
fermented soybean hypocotyl material, 0.05 g of the solid extract
prepared in Example 1-1 by evaporating Liquid Extract 1-2 obtained
using 75Et to dryness.
Example 4-C-3 Coffee Whitener
[0453] A coffee whitener having the following composition was
produced.
TABLE-US-00053 Equol-containing fermented soybean hypocotyl
material (Reference Example 1-3) 0.40 (g) Purified coconut oil
36.00 Sodium casein 4.50 Skimmed milk 5.00 Lecithin 0.05 Trisodium
citrate 0.20 Sucrose fatty acid ester 0.40 Milk flavor 0.10 Water
Balance Total 100.00 g
Example 4-C-4 Coffee Whitener
[0454] A coffee whitener was produced under the same conditions as
in Example 4-C-3, except for using, instead of the equol-containing
fermented soybean hypocotyl material, 0.05 g of the solid extract
prepared in Example 1-1 by evaporating Liquid Extract 1-2 obtained
using 75Et to dryness.
Example 4-C-5 Custard
[0455] Custard having the following composition was produced.
TABLE-US-00054 Equol-containing fermented soybean hypocotyl
material (Reference Example 1-3) 0.40 (Kg) Margarine 20.00 Starch
syrup 5.00 Whole egg 3.00 Cornstarch 4.00 Modified starch 6.00
Lactic protein 2.00 Sugar 30.00 Water Balance Total 100.00 kg
Example 4-C-6 Custard
[0456] Custard was produced under the same conditions as in Example
4-C-5, except for using, instead of the equol-containing fermented
soybean hypocotyl material, 0.05 kg of the solid extract prepared
in Example 1-1 by evaporating Liquid Extract 1-2 obtained using
75Et to dryness.
Example 4-C-7 Custard
[0457] Custard having the following composition was produced.
TABLE-US-00055 Equol-containing fermented soybean hypocotyl
material (Reference Example 1-3) 0.40 (Kg) Carrageenan 0.40 Gelatin
0.50 Cornstarch 4.00 Corn syrup solids 3.00 Granulated sugar 10.00
Glucose 7.00 Skimmed milk 4.00 Whole egg 3.50 Isomerized sugar 7.00
Skimmed sweetened condensed 3.00 milk Unsalted butter 6.50 Water
50.00
Example 4-C-8 Custard
[0458] Custard was produced under the same conditions as in Example
4-7, except for using, instead of the equol-containing fermented
soybean hypocotyl material, 0.05 kg of the solid extract prepared
in Example 1-1 by evaporating Liquid Extract 1-2 obtained using
75Et to dryness.
Example 4-D Dessert
[0459] Hereinafter, specific production examples of the dessert
according to the present invention are given.
Example 4-D-1 Hard Yogurt
[0460] The ingredients of the following composition were
homogenized, and a starter yogurt was added thereto to be
fermented, thereby producing hard yogurt.
TABLE-US-00056 Equol-containing fermented soybean hypocotyl
material (Reference Example 1-3) 0.40 (Kg) Skimmed milk 9.00
Unsalted butter 5.00 Sugar 6.50 Gelling agent 0.60 Water Balance
Total 100.00 kg
Example 4-D-2 Hard Yogurt
[0461] Hard yogurt was produced under the same conditions as in
Example 4-D-1, except for using, instead of the equol-containing
fermented soybean hypocotyl material, 0.05 kg of the solid extract
prepared in Example 1-1 by evaporating Liquid Extract 1-2 obtained
using 75Et to dryness.
Example 4-D-3 Soft Yogurt
[0462] The ingredients of the following composition were
homogenized, and a starter yogurt was added thereto to be
fermented, thereby producing soft yogurt.
TABLE-US-00057 Equol-containing fermented soybean hypocotyl
material (Reference Example 1-3) 0.40 (Kg) Skimmed milk 9.00 Butter
5.25 Sugar 6.50 Locust bean gum 0.30 Water Balance Total 100.00
kg
Example 4-D-4 Soft Yogurt
[0463] Soft yogurt was produced under the same conditions as in
Example 4-D-3, except for using, instead of the equol-containing
fermented soybean hypocotyl material, 0.05 kg of the solid extract
prepared in Example 1-1 by evaporating Liquid Extract 1-2 obtained
using 75Et to dryness.
Example 4-D-5 White Peach Jelly
[0464] A white peach jelly was produced in accordance with a
general production method of a jelly, using the ingredients of the
following composition.
TABLE-US-00058 Equol-containing fermented soybean hypocotyl
material (Reference Example 1-3) 0.40 (part by weight) Sugar 14.40
.kappa.-Carrageenan 0.60 White peach puree 20.00 Water Balance
Total 100.00 parts by weight
Example 4-D-6 White Peach Jelly
[0465] A white peach jelly was produced under the same conditions
as in Example 4-D-5, except for using, instead of the
equol-containing fermented soybean hypocotyl material, 0.05 parts
by weight of the solid extract prepared in Example 1-1 by
evaporating Liquid Extract 1-2 obtained using 75Et to dryness.
Example 4-D-7 Grapefruit Jelly
[0466] A grapefruit jelly was produced in accordance with a general
production method of a jelly, using the ingredients of the
following composition.
TABLE-US-00059 Equol-containing fermented soybean hypocotyl
material (Reference Example 1-2) 0.40 (part by weight) Sugar 18.00
Gellan gum 0.25 Grapefruit juice 20.00 Calcium lactate 0.10 Citric
acid 0.20 Flavor 0.10 Water Balance Total 100.00 parts by
weight
Example 4-D-8 Grapefruit Jelly
[0467] A grapefruit jelly was produced under the same conditions as
in Example 4-D-7, except for using, instead of the equol-containing
fermented soybean hypocotyl material, 0.05 parts by weight of the
solid extract prepared in Example 1-1 by evaporating Liquid Extract
1-2 obtained using 75Et to dryness.
Example 4-D-9 Coffee Jelly
[0468] A coffee jelly was produced in accordance with a general
production method of a jelly, using the ingredients of the
following composition.
TABLE-US-00060 Equol-containing fermented soybean 0.40 (part by
hypocotyl material (Reference Example 1-2) weight) Reducing starch
syrup 10.00 Xylitol 5.00 .kappa.-Carrageenan 0.50 Agar 0.20 Locust
bean gum 0.20 Sugar 3.00 Flavor 0.15 Water Balance Total 100.00
parts by weight
Example 4-D-10 Coffee Jelly
[0469] A coffee jelly was produced under the same conditions as in
Example 4-D-9, except for using, instead of the equol-containing
fermented soybean hypocotyl material, 0.05 parts by weight of the
solid extract prepared in Example 1-1 by evaporating Liquid Extract
1-2 obtained using 75Et to dryness.
Example 4-D-11 Strawberry Milk Jelly
[0470] A strawberry milk jelly was produced in accordance with a
general production method of a jelly, using the ingredients of the
following composition.
TABLE-US-00061 Equol-containing fermented soybean 0.40 (part by
hypocotyl material (Reference Example weight) 1-2) Purified coconut
oil 30.00 Emulsifier 0.25 Skimmed milk 3.30 Sugar 3.50 Reducing
starch syrup 20.00 Lactitol 8.00 Stabilizer 0.70 Xylitol 5.00
Gelatin 0.90 Strawberry puree 5.00 Concentrated strawberry juice
5.00 Citric acid 0.10 Water Balance Total 100.00 parts by
weight
Example 4-D-12 Strawberry Milk Jelly
[0471] A strawberry milk jelly was produced under the same
conditions as in Example 4-D-11, except for using, instead of the
equol-containing fermented soybean hypocotyl material, 0.05 parts
by weight of the solid extract prepared in Example 1-1 by
evaporating Liquid Extract 1-2 obtained using 75Et to dryness.
Example 4-E Snack
[0472] Hereinafter, specific production examples of the snack
according to the present invention are given.
Example 4-E-1 Hard Candy
[0473] A hard candy was produced in accordance with a general
production method of a hard candy, using the ingredients of the
following composition.
TABLE-US-00062 Equol-containing fermented soybean hypocotyl 4.00
(g) material (Reference Example 1-3) Sugar 70.00 Starch syrup 40.00
Water 20.00
Example 4-E-2 Hard Candy
[0474] A hard candy was produced under the same conditions as in
Example 4-E-1, except for using, instead of the equol-containing
fermented soybean hypocotyl material, 0.5 g of the solid extract
prepared in Example 1-1 by evaporating Liquid Extract 1-2 obtained
using 75Et to dryness.
Example 4-E-3 Ginger Candy
[0475] A ginger candy was produced in accordance with a general
production method of a candy, using the ingredients of the
following composition.
TABLE-US-00063 Equol-containing fermented soybean hypocotyl 4.00
(g) material (Reference Example 1-1) Ginger paste 1.50 Reducing
lactose 60.00 Reducing maltose syrup 56.00 Sucralose 0.03 Flavor
0.075 Water 30.00
Example 4-E-4 Ginger Candy
[0476] A ginger candy was produced under the same conditions as in
Example 4-E-3, except for using, instead of the equol-containing
fermented soybean hypocotyl material, 0.5 g of the solid extract
prepared in Example 1-1 by evaporating Liquid Extract 1-2 obtained
using 75Et to dryness.
Example 4-E-5 Throat Soothing Herb Candy
[0477] A throat soothing herb candy was produced in accordance with
a general production method of a candy, using the ingredients of
the following composition.
TABLE-US-00064 Equol-containing fermented soybean hypocotyl 4.00
(g) material (Reference Example 1-2) Mixed herb extract 0.60 Sugar
40.00 Reducing maltose syrup 56.00 Peppermint Flavor 0.20 Caramel
coloring 0.20 Water 20.00
Example 4-E-6 Throat Soothing Herb Candy
[0478] A throat soothing herb candy was produced under the same
conditions as in Example 4-E-5, except for using, instead of the
equol-containing fermented soybean hypocotyl material, 0.5 g of the
solid extract prepared in Example 1-1 by evaporating Liquid Extract
1-2 obtained using 75Et to dryness.
Example 4-E-7 Lemon Gummy Candy
[0479] A lemon gummy candy was produced in accordance with a
general production method of a gummy candy, using the ingredients
of the following composition.
TABLE-US-00065 Equol-containing fermented soybean hypocotyl 4.00
(g) material (Reference Example 1-2) Acid gelatin (250 blooms) 7.00
Reducing starch syrup 100.00 4 times concentrated lemon juice 1.25
Citric acid (crystalline) 1.00 Sucralose 0.02 Food coloring 0.02
Flavor 0.02 Water 12.00
Example 4-E-8 Lemon Gummy Candy
[0480] A lemon gummy candy was produced under the same conditions
as in Example 4-E-7, except for using, instead of the
equol-containing fermented soybean hypocotyl material, 0.5 g of the
solid extract prepared in Example 1-1 by evaporating Liquid Extract
1-2 obtained using 75Et to dryness.
Example 4-E-9 Caramel Candy
[0481] A caramel candy was produced in accordance with a general
production method of a caramel candy, using the ingredients of the
following composition.
TABLE-US-00066 Equol-containing fermented soybean hypocotyl 4.00
(g) material (Reference Example 1-1) Sweetened condensed milk 45.00
Starch syrup 45.00 Vegetable fat and oil 3.50 Lecithin 0.20 Fondant
5.00 Flavor (caramel flavor) 0.10
Example 4-E-10 Caramel Candy
[0482] A caramel candy was produced under the same conditions as in
Example 4-E-9, except for using, instead of the equol-containing
fermented soybean hypocotyl material, 0.5 g of the solid extract
prepared in Example 1-1 by evaporating Liquid Extract 1-2 obtained
using 75Et to dryness.
Example 4-E-11 Pretzel
[0483] The following ingredients were mixed to prepare dough, which
was baked in an oven; thereby, a pretzel was produced.
TABLE-US-00067 Equol-containing fermented soybean 4.00 (g)
hypocotyl material (Reference Example 1-1) Strong flour 100.00
Baking powder 1.35 Unsalted butter 25.00 Table salt 2.00 Sugar
17.50 Soybean dietary fiber 0.80 Sucrose fatty acid ester 0.30
Flavor 0.20 Water 40.00
Example 4-E-12 Pretzel
[0484] A pretzel was produced under the same conditions as in
Example 4-E-11, except for using, instead of the equol-containing
fermented soybean hypocotyl material, 0.5 g of the solid extract
prepared in Example 1-1 by evaporating Liquid Extract 1-2 obtained
using 75Et to dryness.
Example 4-E-13 Mint Tablet
[0485] A mint tablet was produced in accordance with a general
production method of a tablet, using the ingredients of the
following composition.
TABLE-US-00068 Equol-containing fermented soybean hypocotyl 4.00
(g) material (Reference Example 1-3) Sorbitol 90.00 Sucralose 0.15
Peppermint flavor 5.00 Sucrose fatty acid ester 1.00
Example 4-E-14 Mint Tablet
[0486] A mint tablet was produced under the same conditions as in
Example 4-E-13, except for using, instead of the equol-containing
fermented soybean hypocotyl material, 0.5 g of the solid extract
prepared in Example 1-1 by evaporating Liquid Extract 1-2 Obtained
Using 75Et to Dryness.
Example 4-E-15 Yogurt Tablet
[0487] A yogurt tablet was produced in accordance with a general
production method of a tablet, using the ingredients of the
following composition.
TABLE-US-00069 Equol-containing fermented soybean hypocotyl 4.00
(g) material (Reference Example 1-2) Trehalose (granulation) 88.60
Sucralose 0.08 Powdered fermented milk 5.00 Anhydrous citric acid
1.00 Yogurt flavor 0.20 Sucrose fatty acid ester 1.50
Example 4-E-16 Yogurt Tablet
[0488] A yogurt tablet was produced under the same conditions as in
Example 4-E-15, except for using, instead of the equol-containing
fermented soybean hypocotyl material, 0.5 g of the solid extract
prepared in Example 1-1 by evaporating Liquid Extract 1-2 obtained
using 75Et to dryness.
Example 4-E-17 Lemon Tablet
[0489] A lemon tablet was produced in accordance with a general
production method of a tablet, using the ingredients of the
following composition.
TABLE-US-00070 Equol-containing fermented soybean hypocotyl 4.00
(g) material (Reference Example 1-1) Sugar (granulation) 89.50
Powdered lemon juice 2.00 L-ascorbic acid 3.00 Anhydrous citric
acid 1.00 Sucrose fatty acid ester 1.00
Example 4-E-18 Lemon Tablet
[0490] A lemon tablet was produced under the same conditions as in
Example 4-E-17, except for using, instead of the equol-containing
fermented soybean hypocotyl material, 0.5 g of the solid extract
prepared in Example 1-1 by evaporating Liquid Extract 1-2 obtained
using 75Et to dryness.
Example 4-E-19 Mint Chewing Gum
[0491] Mint chewing gum was produced in accordance with a general
production method of a chewing gum, using the ingredients of the
following composition.
TABLE-US-00071 Equol-containing fermented soybean hypocotyl 4.00
(g) material (Reference Example 1-1) Gum base 25.00 Lactitol 63.20
Sugar alcohol 5.00 Sucralose 0.13 Glycerine 0.50 Peppermint oil
1.50 Mint Flavor 0.50 Food coloring 0.20
Example 4-E-20 Mint Chewing Gum
[0492] Mint chewing gum was produced under the same conditions as
in Example 4-E-19, except for using, instead of the
equol-containing fermented soybean hypocotyl material, 0.5 g of the
solid extract prepared in Example 1-1 by evaporating Liquid Extract
1-2 obtained using 75Et to dryness.
Example 4-E-21 Apple Bubble Gum
[0493] Apple bubble gum was produced in accordance with a general
production method of a chewing gum, using the ingredients of the
following composition.
TABLE-US-00072 Equol-containing fermented soybean hypocotyl 4.00
(g) material (Reference Example 1-1) Gum base 30.00 Palatinit 70.00
Sucralose 0.084 Glycerine 0.50 Citric acid (crystalline) 1.00 Apple
flavor 0.80
Example 4-E-22 Apple Bubble Gum
[0494] Apple bubble gum was produced under the same conditions as
in Example 4-E-21, except for using, instead of the
equol-containing fermented soybean hypocotyl material, 0.5 g of the
solid extract prepared in Example 1-1 by evaporating Liquid Extract
1-2 obtained using 75Et to dryness.
Example 4-E-23 Strawberry Chewing Gum
[0495] Strawberry chewing gum was produced in accordance with a
general production method of a chewing gum, using the ingredients
of the following composition.
TABLE-US-00073 Equol-containing fermented soybean hypocotyl 4.00
(g) material (Reference Example 1-3) Gum base 23.00 Powdered sugar
62.00 Starch syrup 5.00 Sucralose 0.001 Citric acid (crystalline)
1.20 Red cabbage color 0.20 Strawberry flavor 0.20
Example 4-E-24 Strawberry Chewing Gum
[0496] Strawberry chewing gum was produced under the same
conditions as in Example 4-E-23, except for using, instead of the
equol-containing fermented soybean hypocotyl material, 0.5 g of the
solid extract prepared in Example 1-1 by evaporating Liquid Extract
1-2 obtained using 75Et to dryness.
Example 4-E-25 Chocolate
[0497] Chocolate was produced in accordance with a general
production method of chocolate, using the ingredients of the
following composition.
TABLE-US-00074 Equol-containing fermented soybean hypocotyl 4.00
(g) material (Reference Example 1-3) Cacao mass 40.00 Cocoa butter
25.00 Reduced powder sugar 50.00 Sucralose 0.03 Lecithin 0.40
Vanillin 0.05
Example 4-E-26 Chocolate
[0498] Chocolate was produced under the same conditions as in
Example 4-E-25, except for using, instead of the equol-containing
fermented soybean hypocotyl material, 0.5 g of the solid extract
prepared in Example 1-1 by evaporating Liquid Extract 1-2 obtained
using 75Et to dryness.
Example 4-E-27 Strawberry-Flavored Chocolate
[0499] Strawberry-flavored chocolate was produced in accordance
with a general production method of chocolate, using the
ingredients of the following composition.
TABLE-US-00075 Equol-containing fermented soybean hypocotyl
material (Reference 4.00 (g) Example 1-3) Cocoa butter 25.00 Hard
butter 7.00 Whole milk powder 27.00 Sugar 41.00 Strawberry flavor
0.20 Beet red 0.50 Sucralose 0.001 Lecithin 0.30 Vanillin 0.02
Example 4-E-28 Strawberry-Flavored Chocolate
[0500] Strawberry-flavored chocolate was produced under the same
conditions as in Example 4-E-27, except for using, instead of the
equol-containing fermented soybean hypocotyl material, 0.5 g of the
solid extract prepared in Example 1-1 by evaporating Liquid Extract
1-2 obtained using 75Et to dryness.
Example 4-E-29 Cookie
[0501] The following ingredients were mixed to prepare dough, which
was baked in an oven to produce a cookie.
TABLE-US-00076 Equol-containing fermented soybean hypocotyl 0.40
(part by weight) material (Reference Example 1-3) Soft flour 48.50
Baking powder 0.30 Unsalted margarine 24.00 Table salt 0.20 Sugar
12.00 Egg yolk 4.70 Trehalose 22.00 Flavor 0.20 Water 40.00
Example 4-E-30 Cookie
[0502] A cookie was produced under the same conditions as in
Example 4-E-29, except for using, instead of the equol-containing
fermented soybean hypocotyl material, 0.05 parts by weight of the
solid extract prepared in Example 1-1 by evaporating Liquid Extract
1-2 obtained using 75Et to dryness.
Example 4-E-31 Skin of Manju (a Bun Filled with Steamed Azuki Bean
Jam)
[0503] The skin of a manju (a bun filled with steamed azuki bean
jam) was produced in accordance with a general production method of
a skin of manju, using the ingredients of the following
composition.
TABLE-US-00077 Equol-containing fermented soybean hypocotyl 4.00
(part by weight) material (Reference Example 1-2) Soft flour 500.00
Baking powder 2.50 Xanthan gum 1.00 Table salt 6.00 Sugar 30.00
Trehalose 30.00 Dry yeast 7.50 Lard 15.00 Water 200.00
Example 4-E-32 Skin of Manju (a Bun Filled with Steamed Azuki Bean
Jam)
[0504] The skin of a manju (a bun filled with steamed azuki bean
jam) was produced under the same conditions as in Example 4-E-31,
except for using, instead of the equol-containing fermented soybean
hypocotyl material, 0.5 parts by weight of the solid extract
prepared in Example 1-1 by evaporating Liquid Extract 1-2 obtained
using 75Et to dryness.
Example 4-E-33 Apple Pie Filling
[0505] An apple pie filling was produced using the following
ingredients.
TABLE-US-00078 Equol-containing fermented soybean hypocotyl 0.40
(part by weight) material (Reference Example 1-1) Apple 200.00
Maltose 150.00 Sucralose 0.10 Lemon juice 10.00 Walnut 60.00 Raisin
60.00 Apple flavor 0.20
Example 4-E-34 Apple Pie Filling
[0506] An apple pie filling was produced under the same conditions
as in Example 4-E-33, except for using, instead of the
equol-containing fermented soybean hypocotyl material, 0.05 parts
by weight of the solid extract prepared in Example 1-1 by
evaporating Liquid Extract 1-2 obtained using 75Et to dryness.
Example 4-E-35 Pound Cake
[0507] The following ingredients were mixed to prepare dough, which
was baked in an oven to produce a pound cake.
TABLE-US-00079 Equol-containing fermented soybean hypocotyl 0.40
(part by weight) material (Reference Example 1-1) Butter 100.00
Reducing maltose syrup 100.00 Sucralose 0.03 Stevia 0.03 Whole egg
100.00 Flavor 0.22 Soft flour 120.00 Baking powder 1.50
Example 4-E-36 Pound Cake
[0508] A pound cake was produced under the same conditions as in
Example 4-E-35, except for using, instead of the equol-containing
fermented soybean hypocotyl material, 0.05 parts by weight of the
solid extract prepared in Example 1-1 by evaporating Liquid Extract
1-2 obtained using 75Et to dryness.
Example 4-E-37 Bracken-Starch Dumpling
[0509] A bracken-starch dumpling was produced using the following
ingredients.
TABLE-US-00080 Equol-containing fermented soybean hypocotyl 0.40
(part by weight) material (Reference Example 1-1) Rice starch 20.00
Sweet potato starch 40.00 Sugar 6.00 Water Balance Total 100.00
parts by weight
Example 4-E-38 Bracken-Starch Dumpling
[0510] A bracken-starch dumpling was produced under the same
conditions as in Example 4-E-37, except for using, instead of the
equol-containing fermented soybean hypocotyl material, 0.05 parts
by weight of the solid extract prepared in Example 1-1 by
evaporating Liquid Extract 1-2 obtained using 75Et to dryness.
Example 4-E-39 Bean Confectionary
[0511] A bean confectionary was produced using the following
ingredients.
TABLE-US-00081 Equol-containing fermented soybean hypocotyl 6.00
(part by weight) material (Reference Example 1-3) Kanbaiko 7.00
(ground toasted rice cake) Sugar 53.00 Kinako (roasted soybean
powder) 4.00 Baking powder q.s. Flavor q.s. Sweetener q.s. Total
100.00 parts by weight
Example 4-E-40 Bean Confectionary
[0512] A bean confectionary was produced under the same conditions
as in Example 4-E-39, except for using, instead of the
equol-containing fermented soybean hypocotyl material, 0.75 parts
by weight of the solid extract prepared in Example 1-1 by
evaporating Liquid Extract 1-2 obtained using 75Et to dryness.
Example 4-E-41 Bean Confectionary
[0513] A bean confectionary was produced using the following
ingredients.
TABLE-US-00082 Equol-containing fermented soybean hypocotyl 12.00
(part by weight) material (Reference Example 1-2) Rice flour 13.00
Sugar 45.00 Kinako (roasted soybean powder) 15.00 Powdered green
tea 15.00 Baking powder q.s. Flavor q.s. Sweetener q.s. Total
100.00 parts by weight
Example 4-E-42 Bean Confectionary
[0514] A bean confectionary was produced under the same conditions
as in Example 4-E-41, except for using, instead of the
equol-containing fermented soybean hypocotyl material, 1.5 parts by
weight of the solid extract prepared in Example 1-1 by evaporating
Liquid Extract 1-2 obtained using 75Et to dryness.
Example 4-E-43 Bean Confectionary
[0515] A bean confectionary was produced using the following
ingredients.
TABLE-US-00083 Equol-containing fermented soybean hypocotyl 18.00
(part by weight) material (Reference Example 1-1) Kanbaiko 15.00
(ground toasted rice cake) Sugar 52.00 Kinako (roasted soybean
powder) 15.00 Powdered green tea 12.00 Baking powder q.s. Flavor
q.s. Sweetener q.s. Total 100.00 parts by weight
Example 4-E-44 Bean Confectionary
[0516] A bean confectionary was produced under the same conditions
as in Example 4-E-43, except for using, instead of the
equol-containing fermented soybean hypocotyl material, 2.25 parts
by weight of the solid extract prepared in Example 1-1 by
evaporating Liquid Extract 1-2 obtained using 75Et to dryness.
Example 4-E-45 Bean Confectionary
[0517] A bean confectionary was produced using the following
ingredients.
TABLE-US-00084 Equol-containing fermented soybean hypocotyl 6.00
(part by weight) material (Reference Example 1-3) Starch 20.00
Sugar 32.00 Kinako (roasted soybean powder) 10.00 Black sesame
seeds 20.00 Baking powder q.s. Flavor q.s. Sweetener q.s. Total
100.00 parts by weight
Example 4-E-46 Bean Confectionary
[0518] A bean confectionary was produced under the same conditions
as in Example 4-E-45, except for using, instead of the
equol-containing fermented soybean hypocotyl material, 0.75 parts
by weight of the solid extract prepared in Example 1-1 by
evaporating Liquid Extract 1-2 obtained using 75Et to dryness.
Example 4-E-47 Nama-Fu Confectionary (Fresh Gluten Cake)
[0519] A nama-fu confectionary (fresh gluten cake) was produced
using the following ingredients.
TABLE-US-00085 Equol-containing fermented soybean hypocotyl 10.00
(part by weight) material (Reference Example 1-3) Gluten 20.00 Rice
flour 15.00 Kinako (roasted soybean powder) q.s. Emulsifier q.s.
Honey q.s. Sorbitol 20.00 Chocolate 30.00
Example 4-E-48 Nama-Fu Confectionary (Fresh Gluten Cake)
[0520] A nama-fu confectionary (fresh gluten cake) was produced
under the same conditions as in Example 4-E-47, except for using,
instead of the equol-containing fermented soybean hypocotyl
material, 1.25 parts by weight of the solid extract prepared in
Example 1-1 by evaporating Liquid Extract 1-2 obtained using 75Et
to dryness.
Example 4-E-49 Wafer (Rich Type)
[0521] A wafer (rich type) was produced in accordance with a
general production method of a wafer, using the following
ingredients.
TABLE-US-00086 Equol-containing fermented soybean hypocotyl 0.40
(part by weight) material (Reference Example 1-3) Wheat flour
100.00 Sugar 2.00 Fats and oils 3.00 Whole egg 3.00 Skimmed milk
2.00 Sodium bicarbonate 0.30 Cow's milk 50.00 Water 100.00
Example 4-E-50 Wafer (Rich Type)
[0522] A wafer (rich type) was produced under the same conditions
as in Example 4-E-49, except for using, instead of the
equol-containing fermented soybean hypocotyl material, 0.05 parts
by weight of the solid extract prepared in Example 1-1 by
evaporating Liquid Extract 1-2 obtained using 75Et to dryness.
Example 4-E-51 Wafer (Plain)
[0523] A wafer (plain) was produced in accordance with a general
production method of a wafer, using the following ingredients.
TABLE-US-00087 Equol-containing fermented soybean hypocotyl 0.40
(part by weight) material (Reference Example 1-3) Wheat flour
100.00 Fats and oils 2.50 Whole egg 0.30 Ammonium carbonate 0.40
Sodium bicarbonate 0.30 Lecithin 0.05 Water 150.00
Example 4-E-52 Wafer (Plain)
[0524] A wafer (plain) was produced under the same conditions as in
Example 4-E-51, except for using, instead of the equol-containing
fermented soybean hypocotyl material, 0.05 parts by weight of the
solid extract prepared in Example 1-1 by evaporating Liquid Extract
1-2 obtained using 75Et to dryness.
Example 4-E-53 Cheese Cracker
[0525] A cheese cracker was produced in accordance with a general
production method, using the following ingredients.
TABLE-US-00088 Equol-containing fermented soybean hypocotyl 0.40
(part by weight) material (Reference Example 1-3) Wheat flour
100.00 Fats and and oils 9.00 Malt extract 1.30 Sodium bicarbonate
0.60 Spice 0.10 Cheese powder 13.00 Sugar 80.00 Table salt 0.90
Ammonium carbonate 0.60 Lecithin 0.05 Water 33.00
Example 4-E-54 Cheese Cracker
[0526] A cheese cracker was produced under the same conditions as
in Example 4-E-53, except for using, instead of the
equol-containing fermented soybean hypocotyl material, 0.05 parts
by weight of the solid extract prepared in Example 1-1 by
evaporating Liquid Extract 1-2 obtained using 75Et to dryness.
Example 4-E-55 Yokan (Azuki Bean Jelly)
[0527] Yokan (azuki bean jelly) was produced in accordance with a
general production method, using the following ingredients.
TABLE-US-00089 Equol-containing fermented soybean hypocotyl 4.00
(part by weight) material (Reference Example 1-3) Agar 7.50
Granulated sugar 320.00 Red bean paste 620.00 Starch syrup 50.00
Water 300.00
Example 4-E-56 Yokan (Azuki Bean Jelly)
[0528] Yokan (azuki bean jelly) was produced under the same
conditions as in Example 4-E-55, except for using, instead of the
equol-containing fermented soybean hypocotyl material, 0.5 parts by
weight of the solid extract prepared in Example 1-1 by evaporating
Liquid Extract 1-2 obtained using 75Et to dryness.
Example 4-E-57 Soft Cookie
[0529] A soft cookie was produced in accordance with a general
production method, using the following ingredients.
TABLE-US-00090 Equol-containing fermented 4.00 (part by weight)
soybean hypocotyl material (Reference Example 1-3) Wheat flour
200.00 Butter 150.00 Sugar 80.00 Whole egg 150.00
Example 4-E-58 Soft Cookie
[0530] A soft cookie was produced under the same conditions as in
Example 4-E-57, except for using, instead of the equol-containing
fermented soybean hypocotyl material, 0.5 parts by weight of the
solid extract prepared in Example 1-1 by evaporating Liquid Extract
1-2 obtained using 75Et to dryness.
Example 4-E-59 Rice Cracker
[0531] A rice cracker was produced in accordance with a general
production method, using the following ingredients.
TABLE-US-00091 Equol-containing fermented 0.40 (part by weight)
soybean hypocotyl material (Reference Example 1-3) Regular rice
100.00 Hot water q.s.
Example 4-E-60 Rice Cracker
[0532] A rice cracker was produced under the same conditions as in
Example 4-E-59, except for using, instead of the equol-containing
fermented soybean hypocotyl material, 0.5 parts by weight of the
solid extract prepared in Example 1-1 by evaporating Liquid Extract
1-2 obtained using 75Et to dryness.
Example 4-E-61 Suhama (Japanese Traditional Confectionary)
[0533] Suhama (Japanese traditional confectionary) was produced in
accordance with a general production method, using the following
ingredients.
TABLE-US-00092 Equol-containing fermented 4.00 (part by weight)
soybean hypocotyl material (Reference Example 1-3) White superior
soft sugar 600.00 Kinako (roasted soybean powder) 400.00 Starch
syrup 130.00 Water 180.00
Example 4-E-62 Suhama (Japanese Traditional Confectionary)
[0534] Suhama (Japanese traditional confectionary) was produced
under the same conditions as in Example 4-E-61, except for using,
instead of the equol-containing fermented soybean hypocotyl
material, 0.5 parts by weight of the solid extract prepared in
Example 1-1 by evaporating Liquid Extract 1-2 obtained using 75Et
to dryness.
Example 4-E-63 Dry Confectionary
[0535] A dry confectionary was produced in accordance with a
general production method, using the following ingredients.
TABLE-US-00093 Equol-containing fermented 4.00 (part by weight)
soybean hypocotyl material (Reference Example 1-3) Wasanbon sugar
500.00 Shitori (for making the ingredients 25.00 wet during the
process) Kanbaiko (ground toasted rice cake) 75.00
Example 4-E-64 Dry Confectionary
[0536] A dry confectionary was produced under the same conditions
as in Example 4-E-63, except for using, instead of the
equol-containing fermented soybean hypocotyl material, 0.5 parts by
weight of the solid extract prepared in Example 1-1 by evaporating
Liquid Extract 1-2 obtained using 75Et to dryness.
Example 4-E-65 Fruit Gummy Candy
[0537] A fruit gummy candy was produced in accordance with a
general production method, using the following ingredients.
TABLE-US-00094 Equol-containing fermented 4.00 (part by weight)
soybean hypocotyl material (Reference Example 1-3) Sugar 30.00
Starch syrup 50.00 Gelatin 7.00 Fruit juice 5.00 Citric acid 1.50
Flavor 0.20
Example 4-E-66 Fruit Gummy Candy
[0538] A fruit gummy candy was produced under the same conditions
as in Example 4-E-65, except for using, instead of the
equol-containing fermented soybean hypocotyl material, 0.5 parts by
weight of the solid extract prepared in Example 1-1 by evaporating
Liquid Extract 1-2 obtained using 75Et to dryness.
Example 4-E-67 Soy Flour-containing Baked Confectionary
[0539] An amount of 1.25 wt. % of equol-containing fermented
soybean hypocotyl material (Reference Example 1-3) and 25 wt. % of
soy flour, as well as raisins, butter, sugar, egg, orange peel,
pineapple, hard-to-digest dextrin, cacao mass, almond slices, cocoa
powder, table salt, baking powder, and flavors were mixed in a
suitable amount. The resultant mixture (100 wt. %) was then formed
into a rectangle (about 1.5 cm.times.1.5 cm.times.10 cm) using a
mold to be baked in an oven at 180.degree. C., thereby producing a
soy flour-containing baked confectionary.
Example 4-E-68 Soy Flour-Containing Baked Confectionary
[0540] A soybean flour-containing baked confectionary was produced
under the same conditions as in Example 4-E-67, except for using,
instead of the equol-containing fermented soybean hypocotyl
material, 0.25 parts by weight of the solid extract prepared in
Example 1-1 by evaporating Liquid Extract 1-2 obtained using 75Et
to dryness.
Example 4-F Seasoning
[0541] Hereinafter, specific production examples of the seasoning
according to the present invention are given.
Example 4-F-1 Barbecue Sauce
[0542] A barbecue sauce having the following composition was
produced.
TABLE-US-00095 Equol-containing fermented 4.00 (Kg) soybean
hypocotyl material (Reference Example 1-3) Reducing starch syrup
20.00 Trehalose 0.50 Mirin 4.00 Soy sauce (dark) 24.00 Apple puree
19.00 Garlic paste 4.50 Ginger paste 4.50 Sesame oil 0.10 Xanthan
gum 0.50 Red pepper powder 0.10 Black pepper powder 0.05 Citric
acid (crystalline) 0.20 Toasted sesame seeds 0.40 Water Balance
Total 100.00 kg
Example 4-F-2 Barbecue Sauce
[0543] A barbecue sauce was produced under the same conditions as
in Example 4-F-1, except for using, instead of the equol-containing
fermented soybean hypocotyl material, 0.5 kg of the solid extract
prepared in Example 1-1 by evaporating Liquid Extract 1-2 obtained
using 75Et to dryness.
Example 4-F-3 Tomato Ketchup
[0544] A tomato ketchup having the following composition was
produced.
TABLE-US-00096 Equol-containing fermented 4.00 (Kg) soybean
hypocotyl material (Reference Example 1-2) Tomato paste 40.00 Wine
vinegar 18.00 Fructose/glucose syrup 5.00 Soy sauce (dark) 24.00
Table salt 2.50 Garlic powder 0.07 Onion powder 0.10 Cinnamon
powder 0.02 Clove powder 0.01 Water Balance Total 100.00 kg
Example 4-F-4 Tomato Ketchup
[0545] A tomato ketchup was produced under the same conditions as
in Example 4-F-3, except for using, instead of the equol-containing
fermented soybean hypocotyl material, 0.5 kg of the solid extract
prepared in Example 1-1 by evaporating Liquid Extract 1-2 obtained
using 75Et to dryness.
Example 4-F-5 Tsuyu (Dipping Sauce)
[0546] Tsuyu (dipping sauce for tempura, i.e. Japanese deep-fried
food) having the following composition was produced.
TABLE-US-00097 Equol-containing fermented 4.00 (Kg) soybean
hypocotyl material (Reference Example 1-2) Sugar 7.00 Soy sauce
(dark) 26.00 Soy sauce (light) 10.00 Fish sauce 5.00 Mirin 10.00
Sodium L-glutamate monohydrate 3.00 Table salt 0.60 Water Balance
Total 100.00 Kg
Example 4-F-6 Tsuyu (Soup)
[0547] Tsuyu (soup) was produced under the same conditions as in
Example 4-F-5, except for using, instead of the equol-containing
fermented soybean hypocotyl material, 0.5 kg of the solid extract
prepared in Example 1-1 by evaporating Liquid Extract 1-2 obtained
using 75Et to dryness.
Example 4-F-7 Dressing
[0548] The aqueous phase and the oil phase of the following
composition were mixed in a weight ratio of 7:3 to produce a
dressing (separate type).
[0549] Composition of Aqueous Phase
TABLE-US-00098 Equol-containing fermented 4.00 (Kg) soybean
hypocotyl material (Reference Example 1-3) Sugar 6.00 Soy sauce
(dark) 20.00 Brewed vinegar (acidity: 10%) 8.50 Fish sauce 3.00
Lemon juice 2.00 Table salt 2.00 Sodium L-glutamate monohydrate
0.20 Red pepper powder 0.05 Toasted sesame seeds 0.30 Xanthan gum
0.10 Water Balance Total 100.00 kg Composition of Oil Phase Sesame
oil 20.00 (Kg) Corn salad oil 79.00 Seasoning flavor 1.00 Total
100.00 kg
Example 4-F-8 Dressing
[0550] A dressing (separate type) was produced under the same
conditions as in Example 4-F-7, except for using, instead of the
equol-containing fermented soybean hypocotyl material, 0.5 kg of
the solid extract prepared in Example 1-1 by evaporating Liquid
Extract 1-2 obtained using 75Et to dryness.
Example 4-F-9 Sesame Dipping Sauce for Shabu-Shabu (Thinly Sliced
Meet and Vegetables Boiled Quickly in Hot Water)
[0551] Sesame dipping sauce for shabu-shabu having the following
composition was produced.
TABLE-US-00099 Equol-containing fermented 0.40 (Kg) soybean
hypocotyl material (Reference Example 1-2) Soy sauce (light) 12.00
Brewed vinegar 11.00 Garlic powder 0.10 Ginger powder 0.10 White
pepper 0.10 Chicken extract 3.00 Yeast extract 3.00 Sesame oil 1.00
Fructose/glucose syrup 8.00 Mirin 15.00 White miso 10.00 Thickener
3.50 Sugar 8.00 Table salt 10.00 Flavor 0.15 Water Balance Total
100.00 kg
Example 4-F-10 Sesame Dipping Sauce for Shabu-Shabu (Thinly Sliced
Meet and Vegetables Boiled Quickly in Hot Water)
[0552] Sesame dipping sauce for shabu-shabu was produced under the
same conditions as in Example 4-F-9, except for using, instead of
the equol-containing fermented soybean hypocotyl material, 0.05 kg
of the solid extract prepared in Example 1-1 by evaporating Liquid
Extract 1-2 obtained using 75Et to dryness.
Example 4-F-11 Barbecue Sauce (Miso Flavor)
[0553] Barbecue sauce (miso flavor) having the following
composition was produced.
TABLE-US-00100 Equol-containing fermented 4.00 (Kg) soybean
hypocotyl material (Reference Example 1-3) Soy sauce (dark) 25.00
Miso 14.00 D-sorbitol 12.00 Table salt 3.20 Garlic powder 3.30
Garlic paste 2.00 Beef extract 1.00 Toasted sesame seeds 0.50
Sodium L-glutamate monohydrate 0.50 Ginger paste 0.40 Amino acid
seasoning 0.01 Thickener 3.50 Sugar 10.00 Water Balance Total
100.00 kg
Example 4-F-12 Barbecue Sauce (Miso Flavor)
[0554] Barbecue sauce (miso flavor) was produced under the same
conditions as in Example 4-F-11, except for using, instead of the
equol-containing fermented soybean hypocotyl material, 0.05 kg of
the solid extract prepared in Example 1-1 by evaporating Liquid
Extract 1-2 obtained using 75Et to dryness.
Example 4-F-13 Snack Seasoning
[0555] A snack seasoning having the following composition was
produced.
TABLE-US-00101 Equol-containing fermented 0.40 (part by weight)
soybean hypocotyl material (Reference Example 1-3) Table salt 14.00
Tricalcium phosphate 3.00 Disodium succinate 0.20 Sodium
L-glutamate monohydrate 7.00 Powdered soy sauce 3.00 Yeast extract
1.50 Onion powder 1.50 Paprika powder 1.50 Garlic powder 6.00 Red
pepper powder 0.50 Chicken consomme 21.00 Beef consomme 4.00 Stevia
0.60 Glucose 13.00 Spice 0.01 Cornstarch 23.00 Sugar 2.00
Example 4-F-14 Snack Seasoning
[0556] A snack seasoning was produced under the same conditions as
in Example 4-F-13, except for using, instead of the
equol-containing fermented soybean hypocotyl material, 0.05 parts
by weight of the solid extract prepared in Example 1-1 by
evaporating Liquid Extract 1-2 obtained using 75Et to dryness.
Example 4-F-15 Emulsified Soy Sauce
[0557] Emulsified soy sauce having the following composition was
produced.
TABLE-US-00102 Equol-containing fermented 0.40 (part by weight)
soybean hypocotyl material (Reference Example 1-2) Salad oil 34.00
Soy sauce (dark) 53.00 Gum Arabic 1.00 Sugar 12.00
Example 4-F-16 Emulsified Soy Sauce
[0558] Emulsified soy sauce was produced under the same conditions
as in Example 4-F-15, except for using, instead of the
equol-containing fermented soybean hypocotyl material, 0.05 parts
by weight of the solid extract prepared in Example 1-1 by
evaporating Liquid Extract 1-2 obtained using 75Et to dryness.
Example 4-F-17 Dressing
[0559] A dressing (emulsified type) having the following
composition was produced.
TABLE-US-00103 Equol-containing fermented 0.40 (part by weight)
soybean hypocotyl material (Reference Example 1-2) Salad oil 35.00
Vinegar 10.00 Sugar 11.00 Table salt 4.00 Xanthan gum 0.30 Water
Balance Total 100.00 parts by weight
Example 4-F-18 Dressing
[0560] A dressing (emulsified type) was produced under the same
conditions as in Example 4-F-17, except for using, instead of the
equol-containing fermented soybean hypocotyl material, 0.05 parts
by weight of the solid extract prepared in Example 1-1 by
evaporating Liquid Extract 1-2 obtained using 75Et to dryness.
Example 4-F-19 Sauce for Kabayaki (Broiled Seafood)
[0561] Sauce for Kabayaki (broiled seafood) having the following
composition was produced.
TABLE-US-00104 Equol-containing fermented 0.40 (part by weight)
soybean hypocotyl material (Reference Example 1-3) D-sorbitol 10.00
Thickener 0.30 Soy sauce (dark) 38.00 Mirin 20.00 Umami extract
2.00 Table salt 1.80 Sodium L-glutamate monohydrate 1.45 Sodium
inosinate 0.025 Sodium guanylate 0.025 Trehalose 15.00 Sugar 6.00
Caramel coloring 0.50 Onion color 0.10 Flavor 0.50 Water Balance
Total 100.00 parts by weight
Example 4-F-20 Sauce for Kabayaki (Broiled Seafood)
[0562] Sauce for Kabayaki (broiled seafood) was produced under the
same conditions as in Example 4-F-19, except for using, instead of
the equol-containing fermented soybean hypocotyl material, 0.05
parts by weight of the solid extract prepared in Example 1-1 by
evaporating Liquid Extract 1-2 obtained using 75Et to dryness.
Example 4-F-21 Seasoning for Squid Delicacies
[0563] A Seasoning for squid delicacies having the following
composition was produced.
TABLE-US-00105 Equol-containing fermented 0.40 soybean hypocotyl
material (part by weight) (Reference Example 1-2) Soy sauce (dark)
20.00 Soy sauce (light) 10.00 Fish sauce 5.00 Mirin 10.00 Brewed
vinegar 2.00 Sugar 38.00 Table salt 1.00 Sodium L-glutamate
monohydrate 2.00 Umami extract 1.40 Disodium succinate 0.20 Garlic
powder 0.05 Water Balance Total 100.00 parts by weight
Example 4-F-22 Seasoning for Squid Delicacies
[0564] A Seasoning for squid delicacies was produced under the same
conditions as in Example 4-F-21, except for using, instead of the
equol-containing fermented soybean hypocotyl material, 0.05 parts
by weight of the solid extract prepared in Example 1-1 by
evaporating Liquid Extract 1-2 obtained using 75Et to dryness.
Example 4-G Retort-packed Food
[0565] Hereinafter, specific production examples of the
retort-packed food according to the present invention are
given.
Example 4-G-1 Retort-Packed Curry
[0566] Retort-packed curry was produced in accordance with a
general production method, using the following composition.
TABLE-US-00106 Equol-containing fermented 0.40 soybean hypocotyl
material (Kg) (Reference Example 1-3) Curry powder 1.80 Onion 7.00
Unsalted butter 4.50 Garlic paste 0.60 Ginger paste 0.50 Wheat
flour 4.50 Fruit chutney 3.00 Tomato paste 0.50 Table salt 0.55
Sodium L-glutamate monohydrate 0.48 Nucleic acid seasoning 0.015
Demi-glace flavor 1.00 Pork extract seasoning 0.50 Bouillon flavor
1.00 Potato 7.00 Carrot 5.00 Beef 10.00 Water Balance Total 100.00
Kg
Example 4-G-2 Retort-Packed Curry
[0567] Retort-packed curry was produced under the same conditions
as in Example 4-G-1, except for using, instead of the
equol-containing fermented soybean hypocotyl material, 0.05 kg of
the solid extract prepared in Example 1-1 by evaporating Liquid
Extract 1-2 obtained using 75Et to dryness.
Example 4-G-3 Retort-Packed Premix of Chinese-Style Donburi
[0568] Retort-packed Premix of Chinese-style Donburi was produced
in accordance with a general production method, using the following
composition.
TABLE-US-00107 Equol-containing fermented soybean 0.40 hypocotyl
material (Reference Example 1-3) (Kg) Soy sauce (light) 72.00 Sake
46.00 Table salt 4.00 Sodium L-glutamate monohydrate 1.30 Nucleic
acid seasoning 0.07 Chicken extract powder 1.00 Sugar 7.00 Water
209.00 Pork 200.00 Squid 120.00 Shrimp 80.00 Shiitake (Lentinus
edodes) 70.00 Boiled bamboo shoots 160.00 Carrot 60.00 Onion
240.00
Example 4-G-4 Retort-Packed Premix of Chinese-Style Donburi
[0569] Retort-packed premix of Chinese-style donburi was produced
under the same conditions as in Example 4-G-3, except for using,
instead of the equol-containing fermented soybean hypocotyl
material, 0.05 kg of the solid extract prepared in Example 1-1 by
evaporating Liquid Extract 1-2 obtained using 75Et to dryness.
Example 8 Processed Meat Product
[0570] Hereinafter, specific production examples of the processed
meat product according to the present invention are given.
Example 4-H-1 Sausage
[0571] A sausage was produced in accordance with a general
production method of a sausage, using the ingredients of the
following composition.
TABLE-US-00108 Meat portion 5 mm-minced pork (foreshank) 70.00 (Kg)
5 mm-minced pork (lard) 10.00 Ice water 20.00 Total 100.00
Auxiliary ingredients kg Equol-containing fermented soybean 0.40
hypocotyl material (Reference Example 1-3) (Kg) Table salt 1.70
Sodium nitrite 0.012 Polyphosphate 0.30 Sodium L-ascorbate 1.00
Sodium casein 0.50 White pepper powder 0.10 Potassium sorbate 0.20
Amino acid seasoning 0.30 Spice mix 0.50
Example 4-H-2 Sausage
[0572] A sausage was produced under the same conditions as in
Example 4-H-1, except for using, instead of the equol-containing
fermented soybean hypocotyl material, 0.05 kg of the solid extract
prepared in Example 1-1 by evaporating Liquid Extract 1-2 obtained
using 75Et to dryness.
Example 4-H-3 Smoked Sausage
[0573] The meat portion and the seasoning having the following
components respectively were mixed at a ratio of 16:1. Using the
resultant mixture, smoked sausages were produced in accordance with
a general production method of a smoked sausage.
TABLE-US-00109 Meat portion Minced pork (foreshank) 85.00 (Kg)
Whole egg 4.00 Cow's milk 3.00 Ice water 7.00 Seasoning
Equol-containing fermented soybean 0.40 hypocotyl material
(Reference (Kg) Example 1-3) Table salt 24.00 Amino acid seasoning
3.20 Spice 8.65 Minced Onion 16.00 Sugar 48.00
Example 4-H-4 Smoked Sausage
[0574] A smoked sausage was produced under the same conditions as
in Example 4-H-3, except for using, instead of the equol-containing
fermented soybean hypocotyl material, 0.05 kg of the solid extract
prepared in Example 1-1 by evaporating Liquid Extract 1-2 obtained
using 75Et to dryness.
Example 4-H-5 Hamburger Steak
[0575] A hamburger steak was produced in accordance with a general
production method, using the following composition.
TABLE-US-00110 Equol-containing fermented soybean 0.40 hypocotyl
material (Reference (part by weight) Example 1-3) Mixture of ground
beef and pork 45.00 Pork 9.00 Onion 12.00 Whole egg 5.00 Granulated
soy protein 9.00 Sodium casein 5.00 Carrageenan 1.50 Locust bean
gum 0.50 Spice 0.30 Yeast extract 0.20 Table salt 1.00 Sugar 1.20
Water-soluble dietary fiber 1.00 Water Balance Total 100.00 parts
by weight
Example 4-H-6 Hamburger Steak
[0576] A hamburger steak was produced under the same conditions as
in Example 4-H-5, except for using, instead of the equol-containing
fermented soybean hypocotyl material, 0.05 parts by weight of the
solid extract prepared in Example 1-1 by evaporating Liquid Extract
1-2 obtained using 75Et to dryness.
Example 4-I Fish Paste
[0577] Hereinafter, specific production examples of the fish paste
according to the present invention are given.
Example 4-I-1 Fried Kamaboko (Fish Paste Cake)
[0578] A fried Kamaboko (fish paste cake) was produced in
accordance with a general production method of a fish paste cake,
using the ingredients of the following composition.
TABLE-US-00111 Equol-containing fermented soybean 0.40 hypocotyl
material (Reference (Kg) Example 1-3) Surimi 50.00 Table salt 1.50
Amino acid seasoning 0.30 Glycine 0.50 Sodium acetate anhydrous
0.30 Potato starch 8.00 Mirin 1.00 Ice water Balance Total 100.00
Kg
Example 4-I-2 Fried Kamaboko (Fish Paste Cake)
[0579] A fried kamaboko (fish paste cake) was produced under the
same conditions as in Example 4-I-1, except for using, instead of
the equol-containing fermented soybean hypocotyl material, 0.05 kg
of the solid extract prepared in Example 1-1 by evaporating Liquid
Extract 1-2 obtained using 75Et to dryness.
Example 4-J Processed Egg Product
[0580] Hereinafter, specific production examples of the processed
egg product according to the present invention are given.
Example 4-J-1 Thick Japanese Omelet
[0581] A thick Japanese omelet was produced in accordance with a
general production method of a thick Japanese omelet, using the
ingredients of the following composition.
TABLE-US-00112 Equol-containing fermented 0.40 soybean hypocotyl
material (part by weight) (Reference Example 1-3) Whole egg 70.00
Table salt 0.40 Starch 2.00 Soup stock 2.00 Sugar 1.20 Sodium
acetate 1.00 Water Balance Total 100.00 parts by weight
Example 4-J-2 Thick Japanese Omelet
[0582] A thick Japanese omelet was produced under the same
conditions as in Example 4-J-1, except for using, instead of the
equol-containing fermented soybean hypocotyl material, 0.05 parts
by weight of the solid extract prepared in Example 1-1 by
evaporating Liquid Extract 1-2 obtained using 75Et to dryness.
Example 4-K Canned Food
[0583] Hereinafter, specific production examples of the canned food
according to the present invention are given.
Example 4-K-1 Canned Mandarin Orange
[0584] A canned mandarin orange was produced by filling mandarin
orange pulp and a syrup having the following composition in a
can
TABLE-US-00113 Equol-containing fermented soybean 0.40 hypocotyl
material (Reference (part by weight) Example 1-3) Sugar 10.00
Fructose/glucose syrup 10.00 Citric acid 0.25 Trisodium citrate
0.10 Water Balance Total 100.00 parts by weight
Example 4-K-2 Canned Mandarin Orange
[0585] Canned mandarin orange was produced under the same
conditions as in Example 4-K-1, except for using, instead of the
equol-containing fermented soybean hypocotyl material, 0.05 parts
by weight of the solid extract prepared in Example 1-1 by
evaporating Liquid Extract 1-2 obtained using 75Et to dryness.
Example 4-L Bread Product
[0586] Hereinafter, specific production examples of the bread
according to the present invention are given.
Example 4-L-1 Loaf of Bread
[0587] A loaf of bread was produced using dough having the
following composition.
TABLE-US-00114 Sponge Dough Strong flour 70.00 (Kg) Yeast food 0.10
Live yeast 2.00 Water 40.00 Main Dough Equol-containing fermented
soybean 0.40 hypocotyl material (Reference (Kg) Example 1-3) Strong
flour 30.00 Sugar 3.00 Table salt 2.00 Skimmed milk 2.00 Shortening
5.00 Water 25.00
Example 4-L-2 Loaf of Bread
[0588] A loaf of bread was produced under the same conditions as in
Example 4-L-1, except for using, instead of the equol-containing
fermented soybean hypocotyl material, 0.05 kg of the solid extract
prepared in Example 1-1 by evaporating Liquid Extract 1-2 obtained
using 75Et to dryness.
Example 4-M Frozen Desert
[0589] Hereinafter, specific production examples of the frozen
dessert according to the present invention are given.
Example 4-M-1 Ice Cream
[0590] Ice cream having the following composition was produced.
TABLE-US-00115 Equol-containing fermented soybean 0.40 hypocotyl
material (Reference (part by weight) Example 1-3) Raw cream 8.80
Butter 5.00 Starch syrup 16.00 Skimmed milk 6.50 Trehalose 7.00
Sugar 5.00 Stabilizer 0.20 Flavor 0.15 Water Balance Total 100.00
parts by weight
Example 4-M-2 Ice Cream
[0591] Ice cream was produced under the same conditions as in
Example 4-M-1, except for using, instead of the equol-containing
fermented soybean hypocotyl material, 0.05 parts by weight of the
solid extract prepared in Example 1-1 by evaporating Liquid Extract
1-2 obtained using 75Et to dryness.
Example 4-N Processed Soybean Food
[0592] Hereinafter, specific production examples of the processed
soybean food according to the present invention are given.
Example 4-N-1 Fermented Soybeans
[0593] An amount of 99.6 parts by weight of steamed soybeans and
0.4 parts by weight of equol-containing fermented soybean hypocotyl
material (Reference Example 1-3) were mixed. The resultant mixture
was then inoculated with Bacillus natto for fermentation; thereby
fermented soybeans were produced.
Example 4-N-2 Fermented Soybeans
[0594] Fermented Soybeans were produced under the same conditions
as in Example 4-N-1, except for using, instead of the
equol-containing fermented soybean hypocotyl material, 0.05 parts
by weight of the solid extract prepared in Example 1-1 by
evaporating Liquid Extract 1-2 obtained using 75Et to dryness.
Example 4-N-3 Tofu
[0595] Tofu having the following composition was produced.
TABLE-US-00116 Equol-containing fermented soybean 0.40 hypocotyl
material (Reference (part by weight) Example 1-3) Soya Milk 99.30
Magnesium chloride 0.20 Potassium sorbate 0.10 Total 100.00 parts
by weight
Example 4-N-4 Tofu
[0596] Tofu was produced under the same conditions as in Example
4-N-3, except for using, instead of the equol-containing fermented
soybean hypocotyl material, 0.05 parts by weight of the solid
extract prepared in Example 1-1 by evaporating Liquid Extract 1-2
obtained using 75Et to dryness.
Example 4-O Cooked Rice Product
[0597] Hereinafter, specific production examples of the cooked rice
product according to the present invention are given.
Example 4-O-1 Rice Gruel
[0598] Rice gruel was produced in accordance with a general
production method of rice gruel, using the ingredients of the
following composition.
TABLE-US-00117 Equol-containing fermented 0.40 soybean hypocotyl
material (part by weight) (Reference Example 1-3) Rice 160.00 Table
salt 1.00 Water 900.00
Example 4-O-2 Rice Gruel
[0599] Rice gruel was produced under the same conditions as in
Example 4-O-1, except for using, instead of the equol-containing
fermented soybean hypocotyl material, 0.05 parts by weight of the
solid extract prepared in Example 1-1 by evaporating Liquid Extract
1-2 obtained using 75Et to dryness.
Example 4-O-3 Steamed Glutinous Rice with Chestnuts
[0600] Steamed glutinous rice with chestnuts was produced in
accordance with a general production method, using the following
ingredients.
TABLE-US-00118 Equol-containing fermented soybean 4.00 hypocotyl
material (Reference (part by weight) Example 1-3) Glutinous rice
1000.00 Peeled chestnut 300.00 Soup stock 230.00 Black sesame seeds
30.00 Salt solution 6.00
Example 4-O-4 Steamed Glutinous Rice with Chestnuts
[0601] Steamed glutinous rice with chestnuts was produced under the
same conditions as in Example 4-O-3 except for using, instead of
the equol-containing fermented soybean hypocotyl material, 0.5
parts by weight of the solid extract prepared in Example 1-1 by
evaporating Liquid Extract 1-2 obtained using 75Et to dryness.
Example 4-P Soup
[0602] Hereinafter, specific production examples of the soup
according to the present invention are given.
Example 4-P-1 Corn Cream Soup
[0603] Corn cream soup was produced in accordance with a general
production method of a soup, using the ingredients of the following
composition.
TABLE-US-00119 Equol-containing fermented soybean 0.40 hypocotyl
material (Reference (part by weight) Example 1-3) Cow's milk 180.00
Corn 190.00 Consomme 20.00 Salad oil 15.00 Table salt q.s. Pepper
q.s. Water 360.00
Example 4-P-2 Corn Cream Soup
[0604] Corn cream soup was produced under the same conditions as in
Example 4-P-1, except for using, instead of the equol-containing
fermented soybean hypocotyl material, 0.05 parts by weight of the
solid extract prepared in Example 1-1 by evaporating Liquid Extract
1-2 obtained using 75Et to dryness.
BRIEF DESCRIPTION OF THE DRAWINGS
[0605] The patent or application file contains at least one drawing
executed in color. Copies of this patent or patent application
publication with color drawing(s) will be provided by the Office
upon request and payment of the necessary fee.
[0606] FIG. 1 is an electropherogram showing the total proteins
contained in equol-containing fermented soybean hypocotyl obtained
in Reference Example 1-1, soybean cotyledon, and the soybean
hypocotyl.
[0607] FIG. 2 is an electropherogram showing the results of the
detection of major allergens (Gym4, Gm30K, and Gm28K) contained in
the equol-containing fermented soybean hypocotyl obtained in
Reference Example 1-1, soybean cotyledon, and the soybean
hypocotyl.
[0608] FIG. 3 is an electropherogram showing the results of the
detection of major allergens (7S globulin mix, oleocine, and
trypsin inhibitor) contained in the equol-containing fermented
soybean hypocotyl obtained in Reference Example 1-1, soybean
cotyledon, and the soybean hypocotyl.
[0609] FIG. 4 shows TLC analysis results indicating the equol
contents of Liquid Extracts 1-2 and 2-2 in Example 1-1.
[0610] FIG. 5 shows TLC analysis results indicating the saponin
contents of Liquid Extracts 1-2 and 2-2 in Example 1-1.
[0611] FIG. 6 shows the TLC analysis results indicating the equol
contents of Liquid Extracts 1-2 and Residue 2 in Example 1-1.
[0612] FIG. 7 shows TLC analysis results indicating the saponin
contents of Liquid Extracts 1-2 and Residue 2 in Example 1-1.
[0613] FIG. 8 shows TLC analysis results of solutions eluted by
silica gel column chromatography in Example 2-1.
* * * * *