U.S. patent application number 15/399350 was filed with the patent office on 2017-05-25 for reduced sugar syrups and methods of making reduced sugar syrups.
This patent application is currently assigned to Tate & Lyle Ingredients Americas, LLC. The applicant listed for this patent is BASF Enzymes LLC, Tate & Lyle Ingredients Americas, LLC. Invention is credited to Andrew Joseph Hoffman, Rohit Medhekar.
Application Number | 20170145526 15/399350 |
Document ID | / |
Family ID | 46003016 |
Filed Date | 2017-05-25 |
United States Patent
Application |
20170145526 |
Kind Code |
A1 |
Medhekar; Rohit ; et
al. |
May 25, 2017 |
REDUCED SUGAR SYRUPS AND METHODS OF MAKING REDUCED SUGAR SYRUPS
Abstract
A reduced sugar syrup having an advantageously low viscosity is
prepared by hydrolysis of starch or starchy material using a
particular type of alpha amylase enzyme which yields a saccharide
distribution having a low DP1-2 and low DP11+ content. The DP4
content of the syrup may be favorably increased by using a
maltotetragenic alpha amylase enzyme in combination with the
aforementioned alpha amylase enzyme. The syrup is useful in the
production of food, beverage, animal feed, animal health and
nutrition, pharmaceutical, and cosmetic compositions and may be
combined with a high intensity sweetener to provide a composition
capable of being substituted for conventional corn syrups.
Inventors: |
Medhekar; Rohit;
(Schaumburg, IL) ; Hoffman; Andrew Joseph;
(Westpoint, IN) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Tate & Lyle Ingredients Americas, LLC
BASF Enzymes LLC |
Hoffman Estates
La Jolla |
IL
CA |
US
US |
|
|
Assignee: |
Tate & Lyle Ingredients
Americas, LLC
Hoffman Estates
IL
BASF Enzymes LLC
La Jolla
CA
|
Family ID: |
46003016 |
Appl. No.: |
15/399350 |
Filed: |
January 5, 2017 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
13752453 |
Jan 29, 2013 |
9540668 |
|
|
15399350 |
|
|
|
|
61592725 |
Jan 31, 2012 |
|
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A23K 20/10 20160501;
A23L 33/10 20160801; A23L 2/60 20130101; A61K 8/60 20130101; A23V
2002/00 20130101; A61K 47/26 20130101; C13K 1/06 20130101; C12P
19/14 20130101; A23L 27/35 20160801; C12P 19/02 20130101; A61P 3/04
20180101; A23L 27/30 20160801 |
International
Class: |
C13K 1/06 20060101
C13K001/06; C12P 19/02 20060101 C12P019/02; A23L 27/30 20060101
A23L027/30; A23L 33/10 20060101 A23L033/10; A23K 20/10 20060101
A23K020/10; C12P 19/14 20060101 C12P019/14; A23L 2/60 20060101
A23L002/60 |
Claims
1-13. (canceled)
14. The syrup of claim 22, wherein the syrup has a viscosity of
less than about 1500 poise at 20.degree. C. when the syrup has a
dry solids content of 80%.
15-21. (canceled)
22. A syrup comprising water and saccharides, the saccharides
having a saccharide distribution of DP1 1-4%; DP2 10-15%; DP3
9-13%; DP4 7-11%; DP5 6-10%; DP6 13-19%; DP7 12-17%; DP8 4-7%; DP9
3-7%; DP10 2-6%; DP11 7-15%; DP11+ 0-4%, the total equaling
100%.
23. A food, beverage, animal feed, animal health and nutrition,
pharmaceutical, or cosmetic product comprising the syrup of claim
22 and at least one food, beverage, animal feed, animal health and
nutrition, pharmaceutical, or cosmetic ingredient.
24. A sweetener product comprising the syrup of claim 22 and at
least one high intensity sweetener.
25. The sweetener product of claim 24, wherein the at least one
high intensity sweetener is selected from the group consisting of
natural high intensity sweeteners.
26. The sweetener product of claim 24, wherein the at least one
high intensity sweetener is selected from the group consisting of
mogrosides and steviol glycosides.
27. The sweetener product of claim 24, wherein the at least one
high intensity sweetener is selected from the group consisting of
synthetic high intensity sweeteners.
28. The sweetener product of claim 24, wherein the at least one
high intensity sweetener is selected from the group consisting of
sucralose, saccharin, cyclamate, acesulfame potassium, neotame and
aspartame.
29. The syrup of claim 22, wherein the syrup has a polydispersity
of not more than 2.
30. The syrup of claim 22, wherein the syrup has a polydispersity
of not more than 1.8.
31. The syrup of claim 22, wherein the syrup has a polydispersity
of not more than 1.6.
32. The syrup of claim 22, wherein the syrup has a dry solids
content of from about 60 weight % to about 80 weight %.
33. The syrup of claim 22, wherein the syrup is clear and liquid at
20-25.degree. C.
34. The syrup of claim 22, wherein the syrup is clear and liquid at
20-25.degree. C., has a dry solids content of from about 60 weight
% to about 80 weight %, has a polydispersity of not more than 1.6,
and has a viscosity of less than about 1500 poise at 20.degree. C.
when the syrup has a dry solids content of 80%.
35. The food, beverage, animal feed, animal health and nutrition,
pharmaceutical, or cosmetic product of claim 23, wherein the food,
beverage, animal feed, animal health and nutrition, pharmaceutical,
or cosmetic product is a product selected from the group consisting
of beverages, baked goods, confectioneries, frozen dairy products,
meats, breakfast cereals, dairy products, condiments, snack bars,
soups, dressings, mixes, prepared foods, baby foods, diet
preparations, peanut butter, syrups, sweeteners, food coatings, pet
foods, animal feed, animal health and nutrition products, dried
fruit, sauces, gravies, jams and jellies.
36. The food, beverage, animal feed, animal health and nutrition,
pharmaceutical, or cosmetic product of claim 23, wherein the food,
beverage, animal feed, animal health and nutrition, pharmaceutical,
or cosmetic product is a food product having the syrup applied
thereon as a coating or layer.
37. The food, beverage, animal feed, animal health and nutrition,
pharmaceutical, or cosmetic product of claim 36, wherein a sweet
topping comprising the syrup is formed on a surface of a baked
good, doughnut, snack bar, cookie or dry cereal.
38. The food, beverage, animal feed, animal health and nutrition,
pharmaceutical, or cosmetic product of claim 23, wherein the food,
beverage, animal feed, animal health and nutrition, pharmaceutical,
or cosmetic product is a food product having a layer comprised of
the syrup present therein and interposed between two other layers.
Description
CROSS-REFERENCE TO RELATED APPLICATION
[0001] This is a continuation application of Allowed U.S.
Non-Provisional application Ser. No. 13/752,453, filed Jan. 29,
2013, assigned U.S. Pat. No. 9,540,668, to issue on Jan. 10, 2017,
which claims priority to U.S. Provisional Application No.
61/592,725, filed Jan. 31, 2012, the disclosures of which are
incorporated herein by reference in their entirety for all
purposes.
FIELD OF THE INVENTION
[0002] The invention relates to syrups useful as food, beverage,
animal feed, animal health and nutrition, and cosmetic ingredients
which are relatively low in sugar content and viscosity, as well as
methods for making such syrups.
BACKGROUND OF THE INVENTION
[0003] Consumer products with high sugar content have come under
wide criticism for their purported links to obesity and associated
health conditions. Consumers are increasingly looking for products
that have low sugar content on their ingredients label. For label
purposes, sugars are explicitly defined as mono or dimeric
carbohydrates (DP1-2, where DP is "degree of polymerization"). To
fulfill a low sugar content need, food companies are actively
investigating ingredient formulations that will reduce the sugar
content of their formulations. Current corn syrups on the market
can have a sugar content as high as 95% (HFCS); typically, the
sugar content ranges from 40% to 95%. A 63 dextrose equivalent (DE)
corn syrup such as Sweetose.RTM. 4300 will have a sugar content of
64-66% and a high maltose syrup can have a sugar content in the
range of 40-60%. These syrups are currently used as bulking agents,
sweeteners, texture modifiers, and viscosity agents and for
moisture control in food applications. For bulking syrups,
viscosity is an important physical property. One method of reducing
sugar content in corn syrups is to substitute the sugars with
higher carbohydrate polymers (DP>11). But this could
significantly change the colligative properties of these syrups,
especially the viscosity. A change in viscosity cannot only
adversely affect the aesthetic value of the food product but also
creates a need for special manufacturing equipment. To avoid these
issues, there is a need for a corn syrup that has substantially
lower sugar content but possesses a viscosity similar to that of a
63 DE corn syrup.
[0004] Some companies are currently using polyols to reduce the
sugar content of syrups. However, polyols can be expensive and many
of them have undesired gastrointestinal side effects.
BRIEF SUMMARY OF THE INVENTION
[0005] The invention provides a method of making a reduced sugar,
lower viscosity syrup, comprising contacting a starch or starchy
material with a first alpha amylase enzyme in an aqueous medium for
a time effective to hydrolyze the starch or starchy material to
provide a reaction product having a saccharide distribution having
a DP1+DP2 content of about 10% to about 25%, a DP3-11 content of
about 70% to about 90%, and a DP11+ content of 0% to about 15%,
wherein the first alpha amylase enzyme is a polypeptide encoded by
a nucleic acid having at least 70%, 71%, 72%, 73%, 74%, 75%, 76%,
77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%,
90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more, or
complete (100%) sequence identity to GenBank Accession No. AF504065
or an amino acid sequence comprising an enzymatically active
fragment of said polypeptide.
[0006] The above-mentioned method may comprise one or more of the
following further steps: filtering the reaction product; contacting
the reaction product with activated carbon; contacting the reaction
product with an ion exchange resin; removing water from the
reaction product to achieve a dry solids content of from about 65%
to about 85%; subjecting the reaction product to ultrafiltration
using a membrane; and/or combining the reaction product with at
least one high intensity sweetener. In one embodiment, the method
comprises the further steps of filtering the reaction product,
contacting the reaction product with activated carbon and an ion
exchange resin, and removing water from the reaction product to
achieve a dry solids content of from about 65% to about 85%. In
another embodiment, the method is conducted such that the starch or
starchy material is not contacted with acid or any enzyme other
than the first alpha amylase enzyme.
[0007] In one aspect of the above-mentioned method, the aqueous
medium initially has a pH of from about 5 to about 7. The
contacting step may, for example, be carried out at a temperature
of from about 75.degree. C. to about 120.degree. C.
[0008] In one embodiment, the above-mentioned method may be
conducted such that a slurry of the starch or starchy material,
aqueous medium and first alpha amylase enzyme is initially heated,
e.g., jet cooked, at a first temperature of from about 100.degree.
C to about 115.degree. C. and then maintained at a second
temperature of from about 80.degree. C. to about 95.degree. C.
[0009] In one embodiment, an amount of the first alpha amylase
enzyme is used in the above-mentioned method which is from about
0.01 to about 0.2 weight % of the amount of starch or starchy
material.
[0010] The starch used in the method may be a corn starch. The
starchy material may be from corn.
[0011] The DP11+ content of the reaction product obtained may, in
one embodiment, be from 0-5%. The saccharides may have a DP4
content of at least about 35%. In one embodiment, the saccharides
have a DP4 content of at least about 35% and a content of less than
about 6% with respect to each of DP5 to DP10.
[0012] The reaction product obtained by the above-mentioned method
may, in one embodiment, have a viscosity of less than about 1500
poise at 20.degree. C. when the reaction product has a dry solids
content of 80%.
[0013] The above-mentioned method may include a step wherein the
starch or starchy material is additionally contacted with a
maltotetragenic alpha amylase (i.e., contacted with a
maltotetragenic alpha amylase as well as the first alpha amylase
enzyme).
[0014] In one embodiment of the above-mentioned method, a slurry of
the starch or starchy material, aqueous medium and first alpha
amylase enzyme is initially jet cooked to provide a liquefied
starch mixture and the liquefied starch mixture is subsequently
contacted with a maltotetragenic alpha amylase. The first alpha
amylase enzyme may be present in the liquefied starch mixture
during the subsequent contacting with the maltotetragenic alpha
amylase. The maltotetragenic alpha amylase may be a variant of a
Pseudomonas saccharophila maltotetraohydrolyase. The
maltotetragenic alpha amylase may be a variant of a wild-type
maltotetraohydrolyase having the amino acid sequence of SEQ ID NO.
2 set forth in WO 2010/132157, comprising:
[0015] (i) a G223E amino acid substitution, and
[0016] (ii) up to 24 additional amino acid deletions, additions,
insertions, or substitutions compared to the amino acid sequence of
SEQ ID NO. 2 set forth in WO 2010/132157; or
[0017] (iii) at least 70% sequence identity to the amino acid
sequence of SEQ ID NO. 2 set forth in WO 2010/132157,
wherein the variant has alpha-amylase activity.
[0018] In one aspect of the invention, the starch or starchy
material is additionally contacted with a maltotetragenic alpha
amylase, such as a variant of a Pseudomonas saccharophilia
maltotetraohydrolyase (i.e., the starch or starchy material is
contacted with a maltotetragenic alpha amylase as well as the first
alpha amylase enzyme). A slurry of the starch or starchy material,
aqueous medium and first alpha amylase enzyme may be initially jet
cooked to provide a liquefied starch mixture and the liquefied
starch mixture is subsequently contacted with the maltotetragenic
alpha amylase. Using a combination of these different enzymes in
this manner achieves hydrolysis of the starch or starchy material
in a manner which helps to decrease or minimize the formation of
sugars (DP1+2) and higher oligosaccharides (DP11+) while increasing
or maximizing the content of DP4 in the resulting syrup.
[0019] In another aspect, the invention provides a method of making
a reduced sugar, lower viscosity syrup, comprising: [0020] (a)
contacting a starch or a starchy material with a first alpha
amylase enzyme in an aqueous medium, optionally under high shear
conditions such as jet cooking, at a first temperature to provide a
fluid mass of gelatinized starch; [0021] (b) maintaining the fluid
mass at a second temperature for a time effective to provide a
reaction product having a saccharide distribution having a DP1+DP2
content of about 10% to about 25%, a DP3-11 content of about 70% to
about 90%, and a DP11+ content of 0% to about 15%, wherein the
first alpha amylase enzyme is a polypeptide encoded by a nucleic
acid having at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%,
79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%,
92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more, or complete (100%)
sequence identity to GenBank Accession No. AF504065 or an amino
acid sequence comprising an enzymatically active fragment of said
polypeptide; and [0022] (c) subjecting the reaction product to one
or more purification or processing steps to provide the reduced
sugar, lower viscosity syrup; [0023] wherein the only type of
enzyme used in the method is alpha amylase.
[0024] Step (a) may be carried out, for example, at a temperature
of 90.degree. C. or greater or 100.degree. C. or greater, e.g.,
about 100.degree. C. to about 115.degree. C., about 104.degree. C.
to about 108.degree. C., or about 107.degree. C. to about
110.degree. C. Step (a) may be carried out, for example, for a time
of from about 2 to about 20 minutes. Step (b) may be carried out,
for example, at a temperature of from about 80.degree. C. to about
100.degree. C., about 80.degree. C. to about 95.degree. C., or
about 90.degree. C. to about 95.degree. C. or a temperature greater
than 90.degree. C. Step (b) may be carried out, for example, for a
time of from about 1.5 to about 5 hours, about 2 to about 4 hours,
or about 3 hours. The second temperature may be lower than the
first temperature.
[0025] The invention also pertains to a syrup comprising water and
saccharides, the saccharides having a saccharide distribution so as
to provide a DP1+DP2 content of about 10% to about 25%, a DP3-11
content of about 70% to about 90%, and a DP11+ content of 0% to
about 15%, wherein the syrup has a viscosity of not more than about
1400 poise at 20.degree. C. when the syrup has a dry solids content
of 80%. In one aspect, the saccharides have a saccharide
distribution so as to provide a DP4 content of at least about 35%
and a content of less than about 6% with respect to each of DP5 to
DP10. In other aspects, the saccharides have a saccharide
distribution so as to provide a DP11+ content of not more than 10%
or not more than 5%.
[0026] Also afforded by the present invention is a food, beverage,
animal feed, animal health and nutrition, pharmaceutical, or
cosmetic product comprising the aforementioned syrup(s) and at
least one food, beverage, animal feed, animal health and nutrition,
pharmaceutical, or cosmetic ingredient.
[0027] Another aspect of the invention relates to a sweetener
product comprising the syrup(s) and at least one high intensity
sweetener.
[0028] In one embodiment, a syrup is provided comprising water and
saccharides, the saccharides having a saccharide distribution of
DP1 1-4%; DP2 10-15%; DP3 9-13%; DP4 7-11%; DP5 6-10%; DP6 13-19%;
DP7 12-17%; DP8 4-7%; DP9 3-7%; DP10 2-6%; DP11 7-15%; DP11+ 0-4%,
the total equaling 100%. A food, beverage, animal feed, animal
health and nutrition, pharmaceutical, or cosmetic product
comprising the aforementioned syrup and at least one food,
beverage, animal feed, animal health and nutrition, pharmaceutical,
or cosmetic ingredient is provided in another embodiment. Still
another aspect provides a sweetener product comprising the
aforementioned syrup and at least one high intensity sweetener.
[0029] Another embodiment of the invention provides a method of
making a reduced sugar, lower viscosity syrup for food, beverage,
animal feed, animal health and nutrition, pharmaceutical, and
cosmetic compositions, comprising: [0030] (a) jet cooking a slurry
of a starch or a starchy material, a first alpha amylase enzyme and
an aqueous medium at a first temperature of from 100.degree. C. to
115.degree. C.; [0031] (b) maintaining the slurry at a second
temperature of from 80.degree. C. to 95.degree. C. for a time
effective to provide a reaction product having a saccharide
distribution having a DP1+DP2 content of 10% to 25%, a DP3-11
content of 70% to 90%, and a DP11+ content of 0% to 15%, wherein
the first alpha amylase enzyme is a polypeptide encoded by a
nucleic acid having at least 70%, 71%, 72%, 73%, 74%, 75%, 76%,
77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%,
90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more, or
complete (100%) sequence identity to GenBank Accession No. AF504065
or an amino acid sequence comprising an enzymatically active
fragment of said polypeptide; and [0032] (c) subjecting the
reaction product to one or more purification or processing steps to
provide the reduced sugar, lower viscosity syrup; [0033] wherein
the only type of enzyme used in the method is alpha amylase.
DESCRIPTION OF THE DRAWING
[0034] FIG. 1 illustrates in schematic form an exemplary embodiment
of the process of the invention.
DETAILED DESCRIPTION OF THE INVENTION
[0035] The term, "DPN", as used herein, refers to the degree of
polymerization, where N is the number of monomeric units (i.e.,
glucose or dextrose units) in the saccharide, thus DPN reflects the
composition of the saccharide. For example, DP1 is a
monosaccharide; DP2 is a disaccharide; DP1+2 is the total of mono-
and di-saccharides; DP3-11 is the total of DP 3 to DP11; and DP11+
is the total of saccharides containing more than 11 monomeric units
per molecule. DPN is expressed as a weight percent of an individual
saccharide on a total saccharide dry weight basis. The DPN
composition of a product was determined using high performance
liquid chromatography (HPLC). Samples were diluted to approximately
5% solids with Milli-Q water and filtered through a 0.45 .mu.m
filter. Twenty microliters of sample was injected. The separation
was accomplished using a Bio-Rad HPX 42A column, a styrene divinyl
benzene resin based column in the silver form coupled with a
refractive index detector. The 42A column is more lightly
cross-linked than the columns used to analyze HFCS. The lower cross
linking gives the resin an open structure, making it more permeable
to higher molecular weight structures. That coupled with the
ligand-ligand reaction between the silver counter ion on the resin
and the hydroxyl groups on the sugars allows separation up to DP12
with a run time of less than 20 minutes. Quantitation is done using
area percent with no response factors since there are few
commercially available pure sugar standards above maltopentose. The
refractive index responses for all these sugars is expected to be
very similar.
[0036] The term, "DS", as used herein, refers to the percent dry
solids of a substance as determined using the computer program,
Refractive Index Dry Substance (RI-DS), Standard Analytical Method
E-54, Corn Refiners Association, 6.sup.th Edition, 1977, E-54, pp.
1-11.
[0037] The term "sugar", as used herein, refers to mono- and/or
di-saccharides.
[0038] The term, "syrup", as used herein, refers to aqueous
solutions of saccharides.
[0039] The term, "viscosity", as used herein, refers to the
resistance of a fluid to flow. The viscosity of a syrup is
typically affected by temperature and solids concentration.
Viscosity is expressed in terms of poise (P) or centipoise (cps) at
a given temperature and a given % DS.
[0040] The syrup preparation method of the present invention
utilizes starch or starchy material as a feedstock or starting
material. Starch or starchy material can be obtained from a number
of different sources using any number of methods routinely
practiced in the art. For example, starch or starchy material can
be obtained from corn (for example, dent corn) or another cereal
feedstock such as rice, wheat, barley, oats, or sorghum through
well-known wet-milling and dry-milling techniques. In wet milling,
corn or other feedstock can be steeped for a period of time and
then ground to separate the germ, which contains the oil, from the
other components. The remaining non-germ material is a slurry that
includes starch, protein (e.g., gluten) and fiber, which can be
separated into different streams. Starch steams also can be
obtained from corn or another starch-rich feedstocks through dry
milling techniques, which also are practiced routinely in the art.
In addition, starch streams can be obtained from a root or tuber
feedstock such as potato or cassava using either wet-milling or
dry-milling processes.
[0041] An advantage of one embodiment of the present invention is
that a starch or starchy material may be directly converted into a
product having a desirable saccharide distribution and viscosity,
as described previously herein, using a single particular type of
enzyme. That is, in one embodiment of the invention the only type
of enzyme used in the method of making the reduced sugar, lower
viscosity syrup is alpha amylase (although, as will be explained
subsequently in more detail, two different alpha amylase enzymes
may be utilized in one embodiment of the present invention). This
contrasts with methods previously known in the art for making
reduced sugar, low viscosity syrups wherein two steps are required
(e.g., a first liquefaction step using an acid or a first type of
enzyme, followed by a second hydrolysis step using a second type of
enzyme).
[0042] Conventionally, an alpha-amylase is used in processes for
making fermentable sugars from starch, whereby the starch is
treated with the alpha-amylase to make a liquefact, which is then
subsequently reacted with a second enzyme, a glucoamylase, to
convert the intermediate liquefact to fermentable sugars (for
example, glucose), see for example U.S. Pat. Nos. 7,273,740,
7,666,633, and 7,659,102. In contrast, another advantage of the
present invention is the novel and unexpected finding that under
appropriate conditions the liquefact that can be generated using an
alpha-amylase having at least 70%, 75%, 80%, 85%, 90%, 95%, or
more, or complete (100%) sequence identity to GenBank Accession No.
AF504065 or an amino acid sequence comprising an enzymatically
active fragment thereof, can be utilized directly for the
production of the reduced sugar, low viscosity syrups of the
present invention. The ability to use a single enzyme to convert a
starch or starchy material to a syrup of the type described herein
simplifies the manufacturing process, thereby reducing costs.
[0043] The first alpha amylase enzyme employed in the present
invention is a polypeptide encoded by a nucleic acid having at
least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%,
82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%,
95%, 96%, 97%, 98%, 99%, or more, or complete (100%) sequence
identity to GenBank Accession No. AF504065 or an amino acid
sequence comprising an enzymatically active fragment of said
polypeptide. Alpha amylase enzymes suitable for such use in the
present invention are known in the art and are described, for
example, in U.S. Pat. Nos. 7,273,740, 7,666,633, and 7,659,102,
each of which is incorporated herein by reference in its entirety
for all purposes. Such alpha amylases can produce liquefaction
products that have a unimodal molecular weight profile of
saccharides centered within the molecular weight range of 1000 to
2000. Sequence ID No. 1 in the aforementioned patents describes a
nucleic acid which encodes a polypeptide useful for practicing the
present invention. The polypeptide used as the first alpha amylase
enzyme in the present invention thus may be a polypeptide encoded
by a nucleic acid having at least 70%, 71%, 72%, 73%, 74%, 75%,
76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%,
89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more, or
complete (100%) sequence identity to Sequence ID No. 1 as set forth
in U.S. Pat. Nos. 7,273,740, 7,666,633, and 7,659,102, or an
enzymatically active fragment thereof. Sequence ID No. 2 in the
aforementioned patents describes an amino acid sequence which may
be present in a polypeptide useful for practicing the present
invention. Thus, a polypeptide which may be used as the first alpha
amylase enzyme in the invention may comprise an amino acid sequence
having at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%,
80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%,
93%, 94%, 95%, 96%, 97%, 98%, 99%, or more, or complete (100%)
sequence identity to Sequence ID No. 2 as set forth in U.S. Pat.
Nos. 7,273,740, 7,666,633, and 7,659,102, or an enzymatically
active fragment thereof. If so desired, the first alpha amylase
enzyme may be immobilized on a suitable support or within a
suitable matrix.
[0044] The amount of first alpha amylase enzyme utilized in the
process of the present invention may be varied and selected
depending upon the desired rate of reaction, reaction conditions
and so forth, but generally will be within the range of from about
0.01 to about 0.2% or from about 0.1 to about 0.2% based on the dry
weight of the starch or starchy material being reacted.
[0045] The starch or starchy material may be combined with water to
form a slurry, typically containing from about 25 weight % to about
40 weight % (e.g., about 32 to about 35 weight %) starch or starchy
material. The pH of the aqueous medium may be adjusted as desired
by the addition of one or more acids or bases. Typically, it will
be desirable for the pH of the aqueous medium to be somewhat (i.e.,
weakly) acidic to neutral, e.g., within the range of from about 4
to about 7 (in one embodiment, from about 5.5 to about 6).
[0046] The process of the present invention may be carried out
using at least two heating stages, wherein in a first heating stage
the aqueous slurry of starch or starchy material and first alpha
enzyme is heated at a relatively high temperature (e.g.,
100.degree. C. or greater) for a relatively short period of time
and subsequently in a second heating stage the aqueous slurry is
heated at a lower temperature than in the first heating stage
(e.g., 80.degree. C. to 95.degree. C.) for a longer period of time
than in the first heating stage. It will often be advantageous to
carry out the first heating stage under conditions effective to
gelatinize or at least partially solubilize the starch. For
example, after combining with the first alpha amylase enzyme, the
slurry may be first subjected to a high shear cooking step wherein
high shear is applied to the starch while the slurry is heated to a
relatively high temperature (e.g., about 90.degree. C. or more or
about 95.degree. C. or more or about 100.degree. C. or more or
about 105.degree. C. or more, but typically not greater than about
115.degree. C.) for a comparatively short period of time (e.g.,
about 2 to about 20 minutes). The high shear cooking step may be
carried under pressure, i.e., at a pressure greater than
atmospheric pressure. For example, a pressure of at least about 5
kg/cm.sup.2 (e.g., about 8 to about 11 kg/cm.sup.2) may be
utilized. Generally, such high shear conditions are selected to be
effective to gelatinize (at least partially solubilize) the starch.
Jet cooking techniques may be used, wherein the slurry is mixed
with steam at high temperature and pressure (i.e., superatmospheric
pressure) while passing through a narrow orifice. The amount of
steam may be controlled such that complete steam condensation is
achieved or, alternatively, the amount of steam may be in excess.
The steam pressure may be from about 5 bar to about 8 bar
(absolute), for example. The intense turbulence resulting from the
near-instantaneous heating of the starch and the passage of steam
through the jet cooker promotes the rupture and dissolution of
starch granules. The viscosity of the slurry is lowered due to the
mechanical shearing of the high molecular weight starch chains. The
starch slurry may thereby be gelatinized and thinned.
[0047] For example, the slurry may be pumped through a steam jet
having a narrow orifice in a jet cooking step to quickly raise the
temperature to between about 100.degree. C. and about 115.degree.
C. (e.g., from about 104.degree. C. to about 108.degree. C. or from
about 107.degree. C. to about 110.degree. C.). The starch is
immediately gelatinized and, due to the presence of the first alpha
amylase, partially depolymerized through random hydrolysis of
glycosidic bonds by the enzyme to a fluid mass which is easily
pumped. In one embodiment, the starch slurry, after being passed
through the steam jet, may be resident in a tail line for a period
of time of from about 5 to about 8 minutes. The fluid mass may then
be transferred to a vessel, such as a stirred tank, wherein
reaction of the starch with the first alpha amylase enzyme may be
continued at a second, somewhat lower temperature (e.g., about
80-95.degree. C.) until the desired saccharide distribution is
achieved. In one embodiment of the invention, the temperature of
the fluid mass is maintained above 90.degree. C. during the second
heating step in order to inhibit the growth of microorganisms.
Typically, the pH of the fluid mass is not adjusted or changed
before proceeding with the second heating step. In such second
heating step, high shear conditions and above-atmospheric pressures
typically are not utilized. For example, the fluid mass may be
stirred or otherwise mixed or agitated under low shear conditions
and atmospheric (ambient) pressure. Generally speaking, increased
reaction times will result in a higher degree of depolymerization,
providing a lower content of DP11+ saccharides and thereby reducing
the viscosity of the resulting syrup. The alpha amylase treatment
thus may be carried out for an amount of time effective to provide
a DP11+ content of, in various embodiments of the invention, not
greater than 15%, not greater than 10%, not greater than 5%, not
greater than 4%, not greater than 3%, not greater than 2%, not
greater than 1%, or approximately 0%. However, it will also
generally be desirable to halt the depolymerization before the
mono- and di-saccharide content becomes unacceptably high. For
example, the enzyme hydrolysis reaction may be stopped when the
DP1+2 content reaches 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%. 18%,
19%, 20%, 21%, 22%, 23%, 24%, or 25%. The reaction time in the
second heating step will typically be about 1.5 to about 5 hours or
about 2 to about 4 hours or about 3 hours.
[0048] According to one embodiment of the invention, the
aforementioned first alpha amylase enzyme is the only enzyme
contacted with the starch (or starchy material) and the
liquefaction product obtained by jet cooking of the starch (or
starchy material). However, in another embodiment the starch (or
starchy material) and/or liquefaction product is additionally
contacted with at least one other enzyme. In particular, the
liquefaction product containing the first alpha amylase enzyme may
combined with a different alpha amylase enzyme such as a
maltotetragenic alpha amylase and then further reacted until a
syrup with the desired saccharide distribution is achieved.
Accordingly, a slurry of the starch (or starchy material), aqueous
medium and first alpha amylase enzyme is initially subjected to
high shear, high temperature conditions (e.g., jet cooked) to
provide a liquefied starch mixture and the liquefied starch mixture
is subsequently contacted with a maltotetragenic alpha amylase. The
first alpha amylase enzyme may be present in the liquefied starch
mixture during the subsequent contacting with the maltotetragenic
alpha amylase. If the maltotetragenic alpha amylase enzyme is not
sufficiently robust to withstand the high temperatures experienced
during jet cooking, it will be desirable to delay combining such
enzyme with the starch until the liquefaction step is completed and
the temperature of the liquefaction product has been lowered to a
temperature (e.g., less than about 80.degree. C.) where the
activity of the maltotetragenic alpha amylase remains high over
time (i.e., it is not inactivated significantly by the heat which
is experienced).
[0049] In the aforementioned embodiment, the maltotetragenic alpha
amylase may be any alpha amylase that selectively or preferentially
produces a high proportion of DP4 oligosaccharide from the
liquefied starch. Such maltotetragenic alpha amylases are well
known in the art and include, for example, a wild-type Pseudomonas
saccharophila maltotetraohydrolyase or a variant thereof. The
maltotetraohydrolyase expressed by Pseudomonas saccharophila is
variously referred to in the art as Amy3A, PSA, SAS, or PS4.
Wild-type Pseudomonas saccharophila maltotetraohydrolyase may be
encoded by a nucleotide sequence as set forth Zhou et al.,
"Nucleotide sequence of the maltotetraohydrolase gene from
Pseudomonas saccharophila," FEBS Lett. 255: 37-41 (1989),
incorporated herein by reference in its entirety. This nucleotide
sequence has been assigned GenBank Accession No. X16732. The
maltotetragenic alpha amylase may be a variant of a wild-type
maltotetraohydrolyase having the amino acid sequence of SEQ ID NO.
2 set forth in WO 2010/132157, comprising:
[0050] (i) a G223E amino acid substitution, and
[0051] (ii) up to 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21,
22, 23 or 24 additional amino acid deletions, additions,
insertions, or substitutions compared to the amino acid sequence of
SEQ ID NO. 2 set forth in WO 2010/132157; or
[0052] (iii) at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%,
79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%,
92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more, or complete (100%)
sequence identity to the amino acid sequence of SEQ ID NO. 2 set
forth in WO 2010/132157, wherein the variant has alpha-amylase
activity. Such variant maltotetragenic alpha amylases are described
in WO 2010/118269 and WO 2010/132157, each of which is incorporated
herein by reference in its entirety for all purposes. Suitable
maltotetragenic alpha amylases are available from commercial
sources, such as certain of the enzymes sold under the
Grindamyl.RTM. PowerFresh brand name by Danisco (Genencor), e.g.,
Grindamyl.RTM. PowerFresh 3001. The maltotetragenic alpha amylase
may be immobilized on a suitable support or within a suitable
matrix, if so desired.
[0053] When a maltotetragenic alpha enzyme is additionally
contacted with the liquefied starch mixture obtained from an
initial jet cooking of starch or starchy material, the pH of the
mixture may typically be maintained within the range of from about
4.5 to about 7. The temperature during such contacting may, for
example, be from about 50 to about 70.degree. C.
[0054] In one embodiment of the invention, the reaction product and
syrup obtained have the following saccharide distribution: DP1
1-4%; DP2 10-15%; DP3 9-13%; DP4 7-11%; DP5 6-10%; DP6 13-19%; DP7
12-17%; DP8 4-7%; DP9 3-7%; DP10 2-6%; DP11 7-15%; DP11+ 0-4%, the
total equaling 100%.
[0055] In another embodiment of the invention, the reaction product
and syrup obtained have the following saccharide distribution: DP1
2-6%; DP2 12-16%; DP3 12-16%; DP4 38-46%; DP5-DP10 1-6% each; DP11
2-10%; DP11+ 0-2%.
[0056] The polydispersity (M.sub.w/M.sub.n) of the saccharides
present in the reaction product and syrup is typically relatively
low, e.g., not more than about 2 or not more than about 1.8 or not
more than about 1.6.
[0057] The saccharide distribution may be monitored on a periodic
basis using methods known in the art and further hydrolysis stopped
by inactivating the enzyme by, for example, adding an amount of
acid effective to lower the pH of the aqueous medium to a level
where the enzyme is no longer active (e.g., a pH of from about 3 to
about 4).
[0058] The reaction product thereby obtained may be subjected to
one or more further purification or processing steps to provide a
syrup suitable for use as a food, beverage, animal feed, or animal
health and nutrition ingredient. For example, the reaction product
may be filtered through a filter medium such as diatomaceous earth
or the like to remove any insoluble substances and/or contacted
with a decolorizing agent such as an ion exchange resin or
activated carbon (typically, by passing the reaction product
through a bed or column packed with the decolorizing agent).
Depending upon the intended end use application, the pH of the
product may be adjusted by addition of acid or base or treatment
with an ion exchange resin. Ultrafiltration using a membrane or the
like may be employed to further reduce the content of mono- and
di-saccharides, if so desired. Water may be removed from the
reaction product by distillation or other evaporative methods to
provide the final syrup having the desired dry solids content and
viscosity. Typically, the dry solids content of the syrup is
advantageously between about 60 weight % and about 85 weight %.
Generally speaking, it will be advantageous to include an amount of
water effective to render the syrup clear and liquid at room
temperature (20-25.degree. C.). In one embodiment of the invention,
the syrup may be dried to provide a solid product (e.g., in the
form of a powder).
[0059] FIG. 1 is a schematic illustration of one embodiment of the
present invention, wherein a slurry of starch is converted using a
single alpha amylase enzyme to a reduced sugar, lower viscosity
syrup by a process involving jet cooking of the starch slurry in
the presence of the alpha amylase followed by continued reaction at
a lower temperature. After deactivation of the enzyme by
acidification, the reaction product is filtered and subjected to
carbon treatment and/or ion exchange prior to being concentrated by
evaporation to provide the final syrup having the desired dry
solids content.
[0060] In certain embodiments of the invention, the syrup may
exhibit the following viscosity profile: [0061] 70% DS: about 12 to
about 22 poise at 20.degree. C., about 5 to about 10 poise at
30.degree. C.; [0062] 75% DS: about 80 to about 110 poise at
20.degree. C., about 30 to about 40 poise at 30.degree. C.; [0063]
80% DS: about 1000 to about 1500 poise at 20.degree. C., about 250
to about 400 poise at 30.degree. C.; [0064] 82% DS: about 4000 to
about 8000 poise at 20.degree. C., about 1000 to about 1600 poise
at 30.degree. C.
[0065] If so desired, the syrup of the present invention may be
converted into dry form by complete or substantially complete
removal of water by any suitable means such as spray-drying.
[0066] The sweetness of the syrup may be increased if so desired by
combining the syrup with one or more high intensity sweeteners of
either natural or synthetic origin. Natural high intensity
sweeteners include, for example, mogrosides and steviol glycosides
(stevia). Illustrative synthetic high intensity sweeteners include
sucralose, saccharin, cyclamate, acesulfame potassium, neotame,
aspartame, and the like. In one embodiment of the invention, an
amount of high intensity sweetener is combined with the syrup to
impart a perceived level of sweetness comparable to that of a
conventional corn syrup having a relatively high content of mono-
and di-saccharides.
[0067] As a result of the low content of polysaccharides (DP11+),
the syrups of the present invention have advantageously low
viscosities at a given dry solids content. This means that the
syrup may be supplied in relatively concentrated form (i.e., a high
DS content) while still having good flow properties, thus
facilitating the incorporation of the syrup into various
foodstuffs. A further advantage of a high DS (low water) content
syrup of the present invention is that it will exhibit improved
microbial stability as compared to a conventional reduced sugar
starch hydrolyzate syrup of comparable viscosity and DP1+2 content,
which necessarily must contain more water due to its higher levels
of DP11+ polysaccharides.
[0068] The syrup of the present invention can be utilized in food,
beverage, animal feed, animal health and nutrition, pharmaceutical,
and cosmetic products to decrease the sugar content of such
products with minimal impact on the physical properties of such
products; and at the same time with minimal impact on the processes
and equipment used for the manufacturing of such products due in
part to the easier handling of such syrup. The syrup may be used in
foods and feeds to soften texture, add volume, thicken, prevent
crystallization of sugar, and/or enhance flavor.
[0069] In particular, the syrup is useful as a bulking agent that
is low in sugar. It is capable of having an appearance, viscosity,
crystallinity, mouthfeel, humectancy and other colligative
properties similar to those of conventional, higher sugar corn
syrups. As such, the syrups of the present invention can be readily
substituted on an approximately equal weight or volume basis for
conventional corn syrups in food, beverage, animal feed, animal
health and nutrition, pharmaceutical, cosmetic products and the
like, yet will effectively reduce the amount of sugar in such
products. The syrups thus can be utilized to lower the sugar
content of products without significantly altering the physical and
sensory attributes of the products. One advantage of the syrups of
the present invention is that they have improved (faster) drying
rates as compared to conventional syrups containing a relatively
high proportion of polysaccharides (e.g., saccharides having a DP
of greater than 11).
[0070] The syrup afforded by the present invention is suitable for
use in food, beverage, animal feed, animal health and nutrition,
pharmaceutical, and cosmetic compositions, especially those which
are reduced sugar or low sugar products. Non-limiting examples
include its use as bulking, binding and coating ingredients;
carriers for coloring agents, flavors/fragrances, and high
intensity sweeteners; spray drying adjuncts; bulking, bodying and
dispersing agents; and ingredients promoting moisture retention
(humectants). Illustrative examples of products which can be
prepared using the syrups described herein include beverages, baked
goods, confectioneries, frozen dairy products, meats, breakfast
cereals, dairy products, condiments, snack bars, soups, dressings,
mixes, prepared foods, baby foods, diet preparations, peanut
butter, syrups, sweeteners, food coatings, pet food, animal feed,
animal health and nutrition products, dried fruit, sauces, gravies,
jams/jellies, and the like.
[0071] The syrup of the present invention may, for example, be
utilized to provide a moisture barrier in various foods. The syrup,
optionally in admixture with one or more additional food
ingredients, may be applied as a coating or layer on a food product
which, once dried, helps to retard the transmission of water into
or out of the food product. For instance, a sweet topping such as a
glaze or frosting comprising the syrup may be formed on a surface
of a baked good such as a doughnut, snack bar, cookie, a dry cereal
(in the form of flakes, biscuits, or clusters, for example) or the
like. The dried sweet topping acts as a moisture barrier, whereby
the resulting food product has improved shelf life and exhibits a
reduced tendency for the outer surface of the sweet topping to
become sticky (tacky) over time. The dried sweet topping can also
hinder the penetration of external moisture into the food product,
thereby permitting the food product to maintain a desired level of
crispness or crunchiness over a prolonged period of time when
immersed in an aqueous environment. In another embodiment, a layer
comprised of the syrup is present within a food product, such that
it is interposed between two other layers (one of which contains
moisture, with the other being lower in moisture content). The
syrup-containing layer helps to slow down or prevent the migration
of moisture from the one layer to the lower moisture content layer.
This serves to maintain the crispiness/crunchiness of the lower
moisture content layer as the food product is stored.
EXAMPLES
Example 1
[0072] 15 kg of starch slurry (35% DS dent starch) was adjusted to
pH 5.8 using 4M NaOH. 5.25 g of Veretase.RTM. enzyme (Verenium
Corporation) (0.1% w/w starch dsb) was added to the slurry. The
slurry was jet cooked at 107.degree. C. with a 6-7 min residence
time in the tail pipe. The jetted starch was collected and allowed
to stir in a round bottom flask maintained at 85-90.degree. C.
Samples were collected for saccharide distribution analysis over
time. The reaction was carried out for 3 hours and then killed by
reducing the pH to 3 and cooling the syrup. The syrup was then
filtered through Celite.RTM. filter aid and passed through
activated carbon and ion exchange resin for purification. The syrup
was then evaporated to 80% DS.
[0073] Table 1A shows the saccharide distribution of the reaction
samples taken at different time intervals. Table 18 provides the
molecular weight and polydispersity data for each sample.
TABLE-US-00001 TABLE 1A Time, min DP1 DP2 DP3 DP4 DP5 DP6 DP7 DP8
DP9 DP10 DP11 DP12 DP13+ 0 0.35 3.06 3.6 2.58 2.16 4.18 6.88 5.36
5.59 4.62 61.56 30 0.81 6.4 6.8 5.23 4.46 8.55 12.56 8.12 6.43 5.42
35.17 60 1.4 8.94 8.73 6.84 5.97 11.62 15.39 8.14 6.03 4.63 4.43
17.78 90 1.69 10.3 9.57 7.6 6.74 13.37 16.13 7.44 5.78 4.62 3.7
13.01 120 1.94 11.27 10.08 8.1 7.27 14.33 16.43 6.77 5.42 3.9 14.44
150 2.09 11.99 10.41 8.5 7.62 15.22 16.23 6.23 5.17 4.04 12.39 180
2.3 12.64 10.73 8.79 8.03 15.8 15.92 5.93 5.11 3.81 10.82 Before
GAC 3.36 14.4 11.81 9.88 9.02 18.04 13.49 5.18 4.38 2.82 7.50 0.00
0.00 IX After GAC 2.81 13.7 11.49 9.75 8.96 17.83 13.55 5.45 4.67
3.21 8.51 0.00 0.00 IX
TABLE-US-00002 TABLE 1B Time, min M.sub.n M.sub.w MP M.sub.z
Polydispersity 0 1139 2025 1500 3342 1.78 30 907 1503 1334 2201
1.66 60 807 1299 1207 1840 1.61 90 747 1198 1120 1701 1.60 120 712
1145 1045 1679 1.61 150 681 1081 1018 1540 1.59 180 650 1030 994
1452 1.59 Final 684 1049 1067 1425 1.53 Final 648 1016 1010 1403
1.57
[0074] The viscosity profile of the syrup thereby obtained at 71%
DS was as shown in Table 2 (compared to SWEETOSE.RTM. 4300 63 DE
conventional corn syrup, 71% DS).
TABLE-US-00003 TABLE 2 Temp., .degree. C. 20 30 40 50 60 70 80
Syrup of the 752 358 190 111 71 51 36 Invention, viscosity in cps
SWEETOSE .RTM. 586 273 142 81 50 33 24 4300, viscosity in cps
Example 2
[0075] 15 kg of starch slurry (35% DS dent starch) was adjusted to
pH 5.3 using 4M NaOH. 5.25 g of Veretase.RTM. enzyme (0.1% w/w
starch dsb) was added to the slurry. The slurry was jet cooked at
107.degree. C. with a 6-7 min residence time in the tail pipe. The
jetted starch was collected and allowed to stir in a round bottom
flask maintained at 85-90.degree. C. Samples were collected for
saccharide distribution analysis over time. The reaction was
carried out for 3 hours and then killed by reducing the pH to 3 and
cooling the syrup. The syrup was then filtered through Celite and
passed through activated carbon and ion exchange resin for
purification. The syrup was then evaporated to 80% DS.
[0076] Table 3 shows the saccharide distribution of the reaction
samples taken at various times.
TABLE-US-00004 TABLE 3 Time, (min) Dextrose DP2 DP3 DP4 DP5 DP6 DP7
DP8 DP9 DP10 DP11 DP12 DP13+ 0 0.56 3.76 4.27 3.2 2.72 5.17 7.98
6.05 5.52 5.21 55.7 30 0.98 6.43 6.81 5.28 4.59 8.8 12.23 8.09 6.47
5.2 4.38 30.7 60 1.28 8.24 8.23 6.48 5.7 10.97 14.66 7.99 6.23 5.09
4.48 20.52 90 1.51 9.47 9.05 7.2 6.33 12.5 15.8 7.5 5.89 4.95 3.61
16.18 120 1.7 10.57 9.68 7.78 7.04 13.78 16.15 7.08 5.53 4.77 3.46
12.53 150 2.03 11.43 10.18 8.23 7.54 14.64 16.13 6.7 5.41 4.26 2.97
10.48 180 kill 2.26 12.25 10.57 8.65 7.9 15.47 15.84 6.33 5.13 4.15
3.05 8.4 (pH 3)
Example 3
[0077] Dent starch (5.25 kg) was mixed with 9.75 kg water to make a
35% DS starch slurry. The pH of the slurry was adjusted to 5.9
using 10% NaOH. 5.25 g of Veretase.RTM. enzyme was added to the
slurry. The slurry was then jet cooked at 107.degree. C. at a rate
of 350 mL/min, which provides a residence time in the tail of 6-7
minutes. The liquefact was collected and cooled to 65.degree. C. in
a water bath. After cooling, 10.5 g of Grindamyl.RTM. PowerFresh
3001 enzyme (Danisco) was added to the syrup. Samples of the
reaction mixture were collected at different time intervals. After
3 hours, the reaction was stopped by reducing the pH to 4. Table 4
shows the saccharide distribution of the reaction samples taken at
different times. The time "t=0" is the time at which jetting
(liquefaction) had been completed and the Grindamyl.RTM. enzyme was
added.
[0078] By way of comparison, when a typical liquefact prepared by
jet cooking a starch slurry using a conventional heat-stable alpha
amylase (e.g., those that produce a bimodal product distribution)
is reacted with Grindamyl.RTM. PowerFresh 3001 enzyme, the reaction
product (syrup) obtained has a relatively high content of DP4
saccharide (e.g., somewhat in excess of 40%). However, the product
also contains a large proportion of higher oligosaccharides (e.g.,
about 30% or more DP11+), which adversely affects the viscosity of
the syrup. The higher oligosaccharides apparently are not
effectively hydrolyzed to lower saccharides by either the
Grindamyl.RTM. PowerFresh enzyme or the conventional alpha amylase
enzyme. The higher oligosaccharides contribute substantially to the
viscosity of the syrup and thus the syrup cannot be used to
effectively replace higher DE syrups, even though it does have a
reduced sugar (DP1+2) content.
TABLE-US-00005 TABLE 4 Sample Dextrose DP2 DP3 DP4 DP5 DP6 DP7 DP8
DP9 DP10 DP11 DP12 DP13+ Veretase .RTM. Liq 0.51 4.12 4.76 3.66
3.15 5.81 8.84 6.58 6.10 4.73 0.00 0.00 51.73 t = 0 0.94 6.29 7.27
8.92 4.50 7.58 10.00 7.13 5.71 5.34 0.00 0.00 36.30 t = 0.5 hr 1.99
9.25 11.13 27.26 4.27 5.57 5.60 7.59 3.93 3.89 0.00 0.00 19.45 t =
1 hr 2.64 10.85 12.52 34.42 3.89 4.89 4.80 6.70 3.39 3.49 2.77 0.00
9.55 t = 1.5 hr 3.05 11.89 13.23 37.99 3.65 4.50 4.35 5.78 3.19
2.90 9.37 0.00 0.00 t = 2 hr 3.33 12.58 13.55 39.76 3.52 4.30 4.12
5.20 3.11 2.64 7.80 0.00 0.00 t = 2.5 hr 3.62 13.31 13.84 41.04
3.42 4.11 4.01 4.68 3.02 2.18 6.65 0.00 0.00 t = 3 hr 3.95 14.08
14.02 41.66 3.42 4.05 3.98 4.28 2.85 2.08 5.51 0.00 0.00
* * * * *