U.S. patent application number 13/874832 was filed with the patent office on 2014-03-06 for extract of a myceliated agricultural substrate and its use as a nutraceutical composition.
This patent application is currently assigned to Mycotechnology, Inc.. The applicant listed for this patent is Brooks John Kelly, Jim Langan, Jonathan Freestone Phillips. Invention is credited to Brooks John Kelly, Jim Langan, Jonathan Freestone Phillips.
Application Number | 20140065131 13/874832 |
Document ID | / |
Family ID | 50187911 |
Filed Date | 2014-03-06 |
United States Patent
Application |
20140065131 |
Kind Code |
A1 |
Kelly; Brooks John ; et
al. |
March 6, 2014 |
EXTRACT OF A MYCELIATED AGRICULTURAL SUBSTRATE AND ITS USE AS A
NUTRACEUTICAL COMPOSITION
Abstract
The present invention provides an aqueous extract of a
myceliated agricultural substrate for use as a nutraceutical or
food composition for human consumption comprising at least one
small molecule compound, wherein mycelia is grown on an
agricultural substrate by inoculating the agricultural substrate
with a pure strain of a fungal culture selected from the group
comprising Basidiomycota and Ascomycota derived from a liquid
fermentation, under optimum growth conditions. The present
invention further provides methods of formation of an aqueous
extract and nutraceutical or food compositions obtained from the
extract. The aqueous extract and the nutraceutical compositions
obtained from the extract thereof have neuroprotective and
neuroregenerative properties amongst many other therapeutic
properties.
Inventors: |
Kelly; Brooks John;
(Longmont, CO) ; Phillips; Jonathan Freestone;
(Longmont, CO) ; Langan; Jim; (Longmont,
CO) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Kelly; Brooks John
Phillips; Jonathan Freestone
Langan; Jim |
Longmont
Longmont
Longmont |
CO
CO
CO |
US
US
US |
|
|
Assignee: |
Mycotechnology, Inc.
Aurora
CO
|
Family ID: |
50187911 |
Appl. No.: |
13/874832 |
Filed: |
May 1, 2013 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
61697506 |
Sep 6, 2012 |
|
|
|
61802256 |
Mar 15, 2013 |
|
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Current U.S.
Class: |
424/115 ;
435/171 |
Current CPC
Class: |
A23G 1/423 20130101;
A23L 33/10 20160801; A23L 31/00 20160801; A23G 1/42 20130101; A23L
21/00 20160801; A23G 1/30 20130101; A23L 5/00 20160801 |
Class at
Publication: |
424/115 ;
435/171 |
International
Class: |
A23L 1/28 20060101
A23L001/28 |
Claims
1. An aqueous extract of a myceliated agricultural substrate for
use as a nutraceutical or food composition for human consumption
comprising at least one small molecule compound, wherein mycelia is
grown on an agricultural substrate by inoculating said agricultural
substrate with a pure strain of a fungal culture selected from the
group comprising Basidiomycota and Ascomycota derived from a liquid
state fermentation, under optimum growth conditions to obtain said
myceliated agricultural substrate, wherein the at least one small
molecule compound enhances bioavailability, water solubility and
passive absorption of the aqueous extract.
2. The extract of claim 1, wherein the at least one small molecule
compound is selected from the group comprising citric acid,
ascorbic acid, honey, dry honey, cane sugar, grape fruit essential
oil, citrus essential oil, tangerine, ethyl alcohol and
combinations thereof.
3. The extract of claim 1, wherein the fungal culture is selected
from the group comprising Hericium erinaceus, Tremella fuciformis,
Agaricus blazei, Cordyceps sinensis, Flammulina velutipes,
Ganoderma lucidum, Grifola frondosa, Letinula edodes, Phellenis
linteus, Polyporus umbellatus, Inonotus obliquus, Pleurotus
ostreatus, P. eryngii, P. citrinopileatus, P. djamor, Trametes
versicolor, Volvariella volvacea, Hypsizygus marmoreus, Pholiota
nameko, Tuber melanosporum, Moschella hortensis, and combinations
thereof.
4. The extract of claim 1, wherein the agricultural substrate is
selected from the group comprising grains, cereals, seeds, dry
fruits, herbs, spices, citrus fruits, tubers, berries, vegetables,
non-legume beans, and combinations thereof.
5. The extract of claim 1, wherein the agricultural substrate is at
least partially coated with vegetable oil, lecithin, and
combination thereof.
6. The extract of claim 1, wherein the extract comprises active
metabolites selected from the group comprising polysaccharides,
glycoproteins, enzymes, proteins, triterpenes, phenols, sterols,
fatty acids, vitamins, mineral salts and combinations thereof.
7. The extract of claim 1, wherein the extract has therapeutic
properties selected from the group comprising anti-aging,
anti-tumor, anti-viral, anti-bacterial, anti-fungal,
hepatoprotective, anti-oxidative, immunomodulating property,
cholesterol regulating property and combinations thereof.
8. The extract of claim 1, wherein the liquid state fermentation
yields spherical conglomerations of said fungal culture wherein
said conglomerations are sized smaller than 2 millimeters in
diameter.
9. The extract of claim 1, wherein said agricultural substrate is
pre-processed before inoculation by treatments selected from the
group comprising heat sterilization, pasteurization, filtration or
combinations thereof.
10. A method of preparation of an aqueous extract of a myceliated
agricultural substrate for use as a nutraceutical composition for
human consumption, comprising the steps of: inoculating an
agricultural substrate with a pure strain of a fungal culture
selected from the group comprising Basidiomycota and Ascomycota,
derived from a liquid fermentation, under optimum growth
conditions; enabling mycelial growth on the agricultural substrate
till the mycelial growth reaches a desired stage to obtain the
myceliated agricultural substrate; obtaining a water-substrate
mixture by boiling the myceliated agricultural substrate in water
for a suitable duration of time and at suitable temperature;
separating aqueous and non-aqueous components from the
water-substrate mixture; and adding at least one small molecule
compound to the aqueous component to form an aqueous extract;
wherein the at least one small molecule compound enhances
bioavailability, water solubility and passive absorption of the
aqueous extract.
11. The method of claim 10, wherein said substrate is pre-processed
by treatments selected from the group comprising heat
sterilization, pasteurization, filtration and combinations
thereof.
12. The method of claim 10, wherein the at least one small molecule
compound is selected from the group comprising citric acid,
ascorbic acid, honey, dry honey, cane sugar, grape fruit essential
oil, citrus essential oil, ethyl alcohol, and combinations
thereof.
13. The method of claim 10, wherein the agricultural substrate is
selected from the group comprising one or more of grains, cereals,
seeds, herbs, spices, dry fruits, berries, citrus fruits, tubers,
vegetables, edible flowers, non-legume beans, and combinations
thereof.
14. The method of claim 10, wherein the liquid media is kept either
stationary or non-stationary during liquid state fermentation.
15. A method of preparation of an aqueous extract of a myceliated
agricultural substrate for use as a nutraceutical composition for
human consumption with neuroprotective and neuroregenerative
activity, comprising the steps of: optionally pre-processing an
agricultural substrate based on nature of the agricultural
substrate; inoculating the agricultural substrate with a pure
strain of a fungal culture selected from a group comprising
Tremella fuciformis and Hericium erinaceus, derived from a liquid
fermentation under optimum growth conditions; enabling mycelial
growth on the agricultural substrate till the mycelial growth
reaches a predetermined stage to obtain the myceliated agricultural
substrate; obtaining a water-substrate mixture by boiling the
myceliated agricultural substrate in water for a suitable duration
of time and at suitable temperature; separating aqueous and
non-aqueous components from the water-substrate mixture; adding at
least one small molecule compound to the aqueous component to form
the aqueous extract; and optionally formulating the aqueous extract
into a nutraceutical or food composition; wherein the at least one
small molecule compound enhances bioavailability, water solubility
and passive absorption of the aqueous extract.
16. The method of claim 15, wherein the at least one small molecule
compound is selected from the group comprising citric acid,
ascorbic acid, honey, cane sugar, grape fruit essential oil, citrus
essential oil, ethyl alcohol and combinations thereof.
17. The method of claim 15, wherein the pre-processing is selected
from the group comprising: a. heat treatment comprising
pasteurization or heat sterilization; b. filtration; c. at least
partial coating of a surface of the agricultural substrate with a
substance selected from the group comprising vegetable oil,
lecithin and combinations thereof.
18. The method of claim 17, wherein the vegetable oil is olive
oil.
19. The method of claim 15, wherein the agricultural substrate is
selected from the group comprising one or more of grains, cereals,
seeds, dry fruits, herbs, spices, citrus fruits, tubers, berries,
vegetables, non-legume beans and combinations thereof.
20. The method of claim 15, wherein the agricultural substrate is
selected from the group comprising green coffee beans, green cocoa
beans and green vanilla beans.
Description
RELATED APPLICATIONS
[0001] This patent application claims the benefit of U.S.
Provisional Patent Application No. 61/697,506, filed 6 Sep. 2012,
and U.S. Provisional Patent Application No. 61/802,256, filed 15
Mar. 2013. This patent application also relates in subject matter
to commonly assigned U.S. patent application Ser. No. 13/859,719,
filed 9 Apr. 2013. The disclosures of each of these related patent
applications are incorporated herein by reference.
FIELD OF THE INVENTION
[0002] The present invention pertains to an extract of a myceliated
agricultural substrate and its use as nutraceutical, food
compositions and products.
BACKGROUND OF THE INVENTION
[0003] Mycelium forms a vegetative part of a fungus consisting of a
network of branched, tubular filaments referred to as hyphal
structures. The mycelial structures play a pivotal role in nutrient
absorption by releasing enzymes into the substrate, including
enzymes for breaking down complex organic polymers into simpler
compounds which are absorbed through hyphal walls.
[0004] Typically a fungal spore germinates into a monokaryotic
mycelia and forms a dikaryotic mycelium on joining with other
compatible mycelia and results in formation of fruiting bodies such
as mushrooms. Mycelium has been used as a primary source for
medicinal extracts and animal food, while the fruiting bodies or
mushrooms are generally used as human food.
[0005] Spore-bearing mushrooms and vegetative mycelia of edible and
medicinal fungal species produce pharmacologically active
metabolites including polysaccharides, proteins, glycoproteins,
enzymes, glucans (alpha, beta), triterpenes, phenols and
sterols.
[0006] It is known from the prior art that mycelial structures and
metabolites of beneficial fungal species possess enhanced
biological activities and numerous therapeutic properties.
Metabolites from more than 650 mushroom species are known to have
anti-tumor and immunomodulatory effects on humans. For instance,
Agaricus blazei is referenced to produce unique alpha and
beta-linked glucans called glucomannan and riboglucan, which are
known anti-viral compounds. A. blazei polysaccharide extracts have
been shown to have anti-cancer activity.
[0007] Pharmaceutical substances with potent health enhancing
properties were isolated recently from medicinal mushrooms and
distributed worldwide. Many of them are pharmaceutical products,
while others represent a novel class of dietary supplements or
"mushroom nutraceuticals". For instance, antitumor polysaccharides,
such as hetero-.beta.-glucans and their protein complexes (e.g.,
xyloglucans, and acidic (.beta.-glucan containing uronic acid) as
well as dietary fiber, lectins, and triterpenoids, have been
isolated from medicinal mushrooms.
[0008] Nutraceutical compositions and food supplements with
therapeutic activity produced from medicinal and edible mushrooms
or fungi have been known in the art. For example, US Patent
Publication US20050180989 discloses treatment of hyperlipemia by
use of an antihyperlipidemic agent and food containing T. matsutake
wherein T. matsutake is provided in the form of mycelia, broth or
fruit bodies including spores; US Patent Publication US2006/0280753
discloses a composition and a method for obtaining a nutritional
food product comprising a UV irradiated, fermented and dried
mycelium/solid substrate mixture; US Patent Publication
US2011/0091579 discloses production of vitamin D2 enriched
mushrooms and fungi for treatment of oxidative stress, Alzheimer's
disease; US Patent Publication US2011/0206721 discloses fermented
soy nutritional supplements including mushroom components and WIPO
publication WO2011/151831 discloses pharmaceutical compositions
comprising extracts of Cyathus Striatus mushrooms.
[0009] Fungi are metabolically similar to animals but structurally
similar to plants in that they possess a rigid cell wall formed
largely of long sugar molecule chains joined by somewhat difficult
to digest beta (b-) linkages and to a smaller extent more easily
digestible alpha (a-) linkages in conjunction with membrane-bound
proteins. In contrast, plant cell walls are made of cellulose
polysaccharides whose 1->4 glycosidic Glucose linkages are
likewise difficult to digest by our enzymes; however, as the cell
walls of Fungi are primarily composed of 1->3 glycosidic
linkages, with 1->6 linked side chains, they may be broken down
by minimal processing using water, heat and mechanical treatment
into smaller, more easily digestible, immunologically-active
polysaccharide molecules of variable microparticulate size called
b-glucans, and related glycoprotein compounds. The immune response
to Glucan is dependent upon a- or b-glucan structure, which has
primary, secondary, and chiral tertiary structures, explaining the
differences in immune response to each Fungi's unique a- and or
b-glucan profile.
[0010] Both mushrooms and mycelium produce polysaccharides and
metabolites, ingestion of which induces positive immunomodulation
in human body. However, most medicinal and/or gourmet mushrooms
take a very long time to come to fruition.
[0011] Thus there is an unmet need in the art to exploit the use of
mycelial tissues instead of fruiting bodies of a variety of
medicinal and edible fungi species, for myceliation and subsequent
extraction to quickly produce extracts of myceliated substrates
where such extracts should be used in formulating nutraceutical
compositions that provide unique properties similar to mushrooms
and or mycelium, including delicious and nutritious meat
substitutes. These types of nutraceuticals should attract consumers
who wish to derive the general immunomodulating, anti-aging,
aphrodisiac, anti-tumour, anti-viral, anti-bacterial, and/or
anti-fungal properties, activities, and benefits, by orally
consuming substrates which has been converted to edible
presentations of a- and b-glucans, glycoproteins, proteins,
ergosterols, sterols, triterpenes, and fatty acids in the form of
nutraceuticals formulations. Furthermore, such new and better
method of preparing an extract of myceliated agricultural material
should be suitable for a variety of mycelial agricultural
substrates in order to produce nutraceutical compositions with
enhanced health benefits.
[0012] Thus the present invention satisfies these unmet needs and
generally overcomes the deficiencies prevalent in the art, by
developing an extract of a myceliated agricultural substrate for
use in formulating nutraceutical compositions, wherein the
compositions offer a variety of health benefits including
neuroregeneration and neuroprotection properties.
SUMMARY OF THE INVENTION
[0013] The present invention relates to an aqueous extract of a
myceliated agricultural substrate for use as a nutraceutical or
food composition for human consumption comprising at least one
small molecule compound, wherein mycelia is grown on an
agricultural substrate by inoculating the agricultural substrate
with a pure strain of a fungal culture selected from a group
comprising Basidiomycota and Ascomycota derived from a liquid
fermentation, under optimum growth conditions, wherein the at least
one small molecule compound enhances bioavailability, water
solubility and passive absorption of the aqueous extract.
[0014] In one embodiment of the present invention, a small molecule
compound can be selected from the group comprising citric acid,
ascorbic acid, honey, cane sugar, grape fruit essential oil, citrus
essential oil, ethyl alcohol and combinations thereof.
[0015] In one embodiment, extract of a myceliated agricultural
substrate of the present invention has neuroprotective and
neuroregenerative properties, wherein fungi strains are selected
from the group comprising Tremella fuciformis and Hericium
erinaceus, further wherein the substrate is pre-processed before
myceliation by at least partially coating the substrate with
lecithin, vegetable oil or combinations thereof.
[0016] The present invention further provides a method of
preparation of an aqueous extract of a myceliated agricultural
substrate for use as a nutraceutical composition for human
consumption, comprising the steps of [0017] inoculating an
agricultural substrate with a pure strain of a fungal culture
selected from the group comprising Basidiomycota and Ascomycota,
derived from a liquid fermentation under optimum growth conditions;
[0018] enabling mycelial growth on the agricultural substrate till
the mycelial growth reaches a desired stage to obtain a myceliated
agricultural substrate; [0019] obtaining a water-substrate mixture
by boiling the myceliated agricultural substrate in water for a
suitable duration of time and at suitable temperature; [0020]
separating aqueous and non-aqueous components from the
water-substrate mixture; [0021] adding at least one small molecule
compound to the aqueous component to form the aqueous extract;
[0022] wherein the at least one small molecule compound enhances
bioavailability, water solubility and passive absorption of the
aqueous extract.
[0023] The present invention further provides a method for
producing a nutraceutical composition with neuroprotective and
neuroregenerative properties from an extract of a myceliated
agricultural substrate comprising the steps of: [0024] optionally
pre-processing an agricultural substrate depending on nature of the
substrate; [0025] inoculating the agricultural substrate with a
pure strain of a fungal culture selected from the group comprising
Tremella fuciformis and Hericium erinaceus, derived from a liquid
fermentation under optimum growth conditions; [0026] enabling
mycelial growth on the agricultural substrate till the mycelial
growth reaches a desired stage to obtain a myceliated agricultural
substrate; [0027] obtaining a water-substrate mixture by boiling
the myceliated agricultural substrate in water for a suitable
duration of time and at a suitable temperature; [0028] separating
aqueous and non-aqueous components from the water-substrate
mixture; [0029] adding at least one small molecule compound to the
aqueous component to form the aqueous extract; and [0030]
optionally formulating the aqueous extract into nutraceutical or
food composition using techniques known in the art;
[0031] wherein the at least one small molecule compound enhances
bioavailability, water solubility and passive absorption of the
aqueous extract.
[0032] In one embodiment, the pre-processing step includes, but not
limited to, for example at least partially coating a surface of the
substrate with lecithin, vegetable oil or combinations thereof.
Preferably the vegetable oil is olive oil.
[0033] In an embodiment, the mycelia from strains of Basidiomycota
and Ascomycota are grown in liquid-state culture and used to
ferment solid-state agricultural substrates.
[0034] Nutraceutical or food composition of the present invention
can further be in a form such as capsule, pill, tablet, syrup,
drink mix, fluid, lozenge, powder and combinations thereof.
BRIEF DESCRIPTION OF THE DRAWINGS
[0035] FIG. 1 is a flowchart of a method for preparing an extract
of myceliated agricultural substrates.
[0036] FIG. 2 is a flowchart of a method of preparation of a
nutraceutical or food composition from an extract of myceliated
agricultural substrates.
DETAILED DESCRIPTION OF THE INVENTION
[0037] The present invention provides an aqueous extract from a
myceliated agricultural substrate for use in formulating
nutraceutical and food compositions, wherein the compositions offer
a variety of health benefits including neuroregeneration and
neuroprotection properties.
[0038] Mycelium extracts have been shown to contain a variety of
pharmacologically important compounds. Utilization of conventional
pharmacological techniques of crystallizing or otherwise
solidifying compounds of interest for delivery in pill form may
diminish or destroy the effectiveness of the compounds by altering
the macromolecular structure, generally done by chemically
modifying polysaccharide structure, for example by way of Smith
degradation, formolysis, and carboxymethylation. Pharmacological
activity of these compounds depends on their molecular structure in
solution.
[0039] The present invention provides mycelia extracts to the human
body fit for oral ingestion in a water-soluble, unaltered form, by
remaining in solution in association with small molecule
compounds.
[0040] FIG. 1 shows a flow chart of a method of preparation of an
extract from myceliated agricultural substrates wherein, [0041]
step 10 provides an agricultural substrate; [0042] step 12
optionally pre-processes the substrate based on nature of the
substrate, [0043] step 14 inoculates an agricultural substrate with
pure culture of fungal strains selected from Basidiomycota and
Ascomycota; [0044] step 16 enables mycelium growth on the substrate
under optimum growth conditions to obtain a myceliated agricultural
substrate; [0045] step 18 grows the mycelium on the myceliated
agricultural substrate till the mycelial growth reaches a desired
stage; [0046] step 20 obtains a water substrate mixture by boiling
the myceliated agricultural substrate with water; [0047] step 22
separates aqueous and non-aqueous components from the water
substrate mixture; and [0048] step 24 adds at least one small
molecule component to the aqueous component to form the aqueous
extract of the myceliated substrate.
[0049] The present invention further provides a method of preparing
a nutraceutical composition from an extract of myceliated
agricultural substrates as shown in FIG. 2, wherein, [0050] step 30
provides an agricultural substrate, [0051] step 32 optionally
pre-processes the substrate based on nature of the substrate,
[0052] step 34 inoculates the substrate with pure culture of fungal
strains selected from Tremella fuciformis and Hericium erinaceus,
[0053] step 36 enables mycelium growth on the substrate under
optimum growth conditions till a desired stage is reached to obtain
a myceliated substrate; [0054] step 38 obtains a water substrate
mixture by boiling the myceliated substrate with water; [0055] step
40 separates aqueous and non-aqueous components from the water
substrate mixture; [0056] step 42 adding at least one small
molecule compound to the aqueous extract; and [0057] step 44
optionally formulates the aqueous extract into a nutraceutical or
food composition using techniques known in the art.
[0058] In an embodiment, optimum growth conditions for mycelial
growth can be controlled by controlling parameters such as
selective pH, temperature, humidity, incubation period, media
composition, aeration and pressure among such other parameters.
[0059] Enabling of mycelia growth on a substrate can further
comprise of maintaining liquid media under stationary condition or
mixing the liquid media by utilizing mixing methods such as
agitation, stirring, swirling continuously or intermittently among
such other mixing methods.
[0060] In an embodiment, the desired stage of mycelia growth can be
reached, for example, when fruiting bodies begin to appear on a
surface of an agricultural substrate.
[0061] In one embodiment, the pre-processing step is selected from
the group comprising:
a. heat treatment including pasteurization or heat sterilization,
b. filtration, c. at least partial coating of a surface of the
substrate with at least one myceliation enhancing compound selected
from a vegetable oil such as olive oil, d. at least partial coating
of a surface of the substrate with at least one substance known to
exhibit neuroregenerative and neuroprotective properties such as
lecithin.
[0062] In one embodiment, aqueous component, obtained after boiling
a myceliated substrate with water, contains polysaccharides and
other beneficial components suspended in water. The non-aqueous
component contains substrate solids. Separation of aqueous and
non-aqueous components can be accomplished by methods such as
filtering or siphoning or other methods known in the art.
[0063] In an embodiment of the present invention, the small
molecule compound is selected from the group comprising citric
acid, ascorbic acid, honey, cane sugar, grape fruit essential oil,
citrus essential oil, ethyl alcohol and combinations thereof,
wherein an addition of the small molecule compound facilitates
water solubility, bioavailability and passive absorption of the
aqueous extract.
[0064] In an embodiment, the agricultural substrate is inoculated
by liquid media comprising an aliquot of pure culture of a fungal
strain selected from Basidiomycota and Ascomycota, wherein the
fungal strains are derived from a liquid state fermentation.
[0065] In a further embodiment, the liquid media can be kept either
stationary or non-stationary during liquid state fermentation. The
liquid media can be stationary creating a semi-anaerobic
environment that enhances a direct mycelial growth downward from a
floating mass into the liquid media. In another embodiment, the
liquid media is mixed by methods such as agitation, swirling and
stirring.
[0066] In one embodiment, mycelia from strains, selected from
Basidiomycota and Ascomycota Fungi, is grown in liquid-state
culture and used to ferment solid-state agricultural
substrates.
[0067] In a yet another embodiment, liquid media can be injected
with sterile air to deliver oxygen to the liquid media.
[0068] In a further embodiment, liquid state fermentation yields
spherical conglomerations of culture that are sized to
interstitially penetrate the substrate to optimize inoculation of
an agricultural substrate. Spherical conglomerations are sized
smaller than 2 millimeters in diameter, preferably between 10
microns to 1 millimeter in diameter.
[0069] In another embodiment of the present invention, addition of
a small molecule compound enables passive absorption of aqueous
extract through lining of oral cavity, oesophagus, stomach and
upper portions of small intestine. Addition of small molecule
compound further facilitates active absorption of unabsorbed
components, if any, at distal end of the small intestine.
[0070] In another embodiment of the present invention, extracts
prepared from myceliated agricultural substrates and associated
with one or more small molecule compound(s), further comprise of
honey. These extracts with honey or dry honey can form a novel,
water-soluble delivery system for delivering pharmacologically
active compounds, which are quickly metabolized. The term "dry
honey" as used herein means bee honey whose physical state is
somewhat more solid than liquid at room temperature. Mycelia
extracts of the present invention have low glycemic index, which
adds health-care benefits to the honey formulations. This
low-glycemic effect can be enhanced by manipulating the honey
formulations into pills, capsules and the like.
[0071] Specific Fungi strain selection and growing of strains
organically is important for maximizing production of
immunologically active-polysaccharides and metabolites from
agricultural substrate(s). In an embodiment, the pure culture of
fungal strain can be selected from a group comprising Hericium
erinaceus, Tremella fuciformis, Agaricus blazei, Cordyceps
sinensis, Flammulina velutipes, Ganoderma lucidum, Grifola
frondosa, Letinula edodes, Phellenis linteus, Polyporus umbellatus,
Inonotus obliquus, Pleurotus ostreatus, P. eryngii, P.
citrinopileatus, P. djamor, Trametes versicolor, Volvariella
volvacea, Hypsizygus marmoreus, Pholiota nameko, Tuber
melanosporum, Moschella hortensis, and combinations thereof.
[0072] In an embodiment, agricultural substrate is selected from
the group comprising one or more of grains, cereals, seeds,
non-legume beans, tubers, citrus fruits, berries, dry fruits,
spices, herbs, vegetables and combinations thereof.
[0073] Agricultural substrates of the present invention can be
pre-processed before inoculation. Pre-processing depends on nature
of the substrate, wherein the pre-processing can comprise heat
treatment including pasteurization and heat sterilization and/or
filtration, based on nature of the substrate in order to eradicate
or minimize microbial load present on its surface.
[0074] In one exemplary embodiment, agricultural substrates such as
grains, cereals, and seeds, prior to inoculation with pure culture
of fungal species, are subjected to heat sterilization treatment
comprising pressure cooking for 90 minutes at 15 lb/inch square in
order to remove microbial content present on the substrate without
adversely affecting its taste and nutritional quality.
[0075] In another exemplary embodiment, agricultural substrates
such as herbs, spices, dry fruits, vanilla beans, coffee beans can
be pasteurized by placing in glass jars or plastic growth bags and
subjected to dry heat treatment at 145 degrees F. to 195 degrees F.
for 30 to 90 minutes in covered stainless steel trays.
[0076] In a further exemplary embodiment, substrates such as honey
and aqueous solution of substrate are subjected to filtration in
order to remove microbial content prior to inoculation with pure
strains of fungal culture.
[0077] In accordance with the present invention, when an
agricultural substrate has substrate elements defining interstitial
spaces in between such as rice, the substrate can be pre-processed
before inoculation by at least partially coating a surface of the
substrate with at least one myceliation enhancing compound
including but not limiting to vegetable oil such as olive oil.
Vegetable oil can increase interstitial spaces between substrate
elements to improve myceliation. Preferably, vegetable oil can be
olive oil, which may have neuro-protectant properties. Further, the
substrate can also be coated with at least one substance known to
exhibit neuroregenerative and/or neuroprotective properties such as
lecithin.
[0078] In an embodiment, present invention provides a method of
preparing a nutraceutical composition with neuroregenerative
properties from an extract of myceliated agricultural substrates,
wherein inoculation of the substrate is carried out using pure
culture of Hericium erinaceus and Tremella fuciformis.
Neuroregenerative properties of Hericium erinaceus and Tremella
fuciformis are known in the art. Moreover, it is also known in the
art that mycelial structures of H. erinaceus show presence of
bioactive compounds with improved neuroregenerative effects than
the compounds present in the fruiting bodies of H. erinaceus.
[0079] In another embodiment, present invention provides an extract
of myceliated agricultural substrate with neuroprotective
activities, wherein the substrate is at least partially coated with
vegetable oil, lecithin or combinations thereof. In one preferred
embodiment, vegetable oil is olive oil. Pre-processing of the
agricultural substrate with lecithin enhances neuroprotective
properties of the extract.
[0080] In one embodiment, surface of the substrate can be coated
with a mixture of vegetable oil and lecithin before the substrate
is inoculated with a pure strain of fungal strain comprising
Hericium erinaceus and Tremella fuciformis followed by myceliation
and subsequent extraction steps as described herein to produce
nutraceutical compositions with enhanced neuroregenerative and/or
neuroprotective properties.
[0081] In an embodiment, extract of the present invention can be
prepared from a water substrate mixture, wherein the water
substrate mixture is obtained by boiling the myceliated substrate
in water at a suitable temperature range for a suitable duration of
time.
[0082] In an alternate embodiment, extract of the present invention
can be prepared by mixing myceliated substrate with specific amount
of solvents such as ethyl alcohol, ethyl acetate, diethyl ether and
incubated for a particular period of time in order to form solvent
based extract. In a further embodiment, the solvent based extract
can be subjected to freeze drying in order to remove aqueous
solvent and organic solvent from the extract.
[0083] In an embodiment, aqueous extract of the present invention
can be separated from water substrate mixture using separation
methods such as filtration, siphoning, centrifugation, vortex
sedimentation, and agitation, heating among such separation
methods.
[0084] Agricultural substrates that are myceliated in accordance
with this invention can include one or more of cereals, grains,
species of wheat, rye, brown rice, white rice, red rice, gold rice,
wild rice, rice, barley, triticale, rice, sorghum, oats, millets,
quinoa, buckwheat, fonio, amaranth, teff and durum; apples and
pears, apricots, cherries, almonds, peaches, strawberries, raisins,
manioc, cocoa, banana, Rubiaceae sp. (coffee), lemons, oranges and
grapefruit; tomatoes, potatoes, peppers, eggplant, Allspice, mango
powder, Angelica, Anise (Pimpinella anisum), Aniseed myrtle
(Syzygium anisatum), Annatto (Bixa orellana), Apple mint (Mentha
suaveolens), Artemisia vulgaris, Mugwort, Asafoetida (Ferula
assafoetida), Berberis, Banana, Basil (Ocimum basilicum), Bay
leaves, Bistort (Persicaria bistorta), Black cardamom, Black cumin,
Blackcurrant, Black limes, Bladder wrack (Fucus vesiculosus), Blue
Cohosh, Blue-leaved Mallee (Eucalyptus polybractea), Bog Labrador
Tea (Rhododendron groenlandicum), Boldo (Peumus boldus), Bolivian
Coriander (Porophyllum ruderale), Borage (Borago officinalis),
Calamus, Calendula, Calumba (Jateorhiza calumba), Chamomile,
Cannabis, Caper (Capparis spinosa), Caraway, Cardamom, Carob Pod,
Cassia, Casuarina, Catnip, Cat's Claw, Catsear, Cayenne pepper,
Celastrus Paniculatus, Comfrey, Celery salt, Celery seed, Centaury,
Chervil (Anthriscus cerefolium), Chickweed, Chicory, Chile pepper,
Chili powder, Cinchona, Chives (Allium schoenoprasum), Cicely
(Myrrhis odorata), Cilantro (see Coriander) (Coriandrum sativum),
Cinnamon (and Cassia), Cinnamon Myrtle (Backhousia myrtifolia),
Clary, Cleavers, Clover, Cloves, Coltsfoot, Comfrey, Common Rue,
Condurango, Coptis, Coriander, Costmary (Tanacetum balsamita),
Couchgrass, Cow Parsley (Anthriscus sylvestris), Cowslip, Cramp
Bark (Viburnum opulus), Cress, Cuban Oregano (Plectranthus
amboinicus), Cudweed, Cumin, Curry leaf (Murraya koenigii), Damiana
(Turnera aphrodisiaca), Dandelion (Taraxacum officinale),
Demulcent, Devil's claw (Harpagophytum procumbens), Dill seed, Dill
(Anethum graveolens), Dorrigo Pepper (Tasmannia stipitata),
Echinacea, Echinopanax Elatum, Edelweiss, Elderberry, Elderflower,
Elecampane, Eleutherococcus senticosus, Epazote (Chenopodium
ambrosioides), Ephedra, Eryngium foetidum, Eucalyptus, Fennel
(Foeniculum vulgare), Fenugreek, Feverfew, Figwort, Five-spice
powder (Chinese), Fo-ti-tieng, Fumitory, Galangal, Garam masala,
Garden cress, Garlic chives, Garlic, Ginger (Zingiber officinale),
Ginkgo biloba, Ginseng, Ginseng, Siberian (Eleutherococcus
senticosus), Goat's Rue (Galega officinalis), Goada masala, Golden
Rod, Golden Seal, Gotu Kola, Grains of paradise (Aframomum
melegueta), Grains of Selim (Xylopia aethiopica), Grape seed
extract, Green tea, Ground Ivy, Guaco, Gypsywort, Hawthorn
(Crataegus sanguines), Hawthorne Tree, Hemp, Herbes de Provence,
Hibiscus, Holly, Holy Thistle, Hops, Horehound, Horseradish,
Horsetail (Equisetum telmateia), Hyssop (Hyssopus officinalis),
Jalap, Jasmine, Jiaogulan (Gynostemma pentaphyllum), Joe Pye weed
(Gravelroot), John the Conqueror, Juniper, Kaffir Lime Leaves
(Citrus hystrix, C. papedia), Kaala masala, Knotweed, Kokam,
Labrador tea, Lady's Bedstraw, Lady's Mantle, Land cress, Lavender
(Lavandula spp.), Ledum, Lemon Balm (Melissa Officinalis), Lemon
basil, Lemongrass (Cymbopogon citratus, C. flexuosus, and other
species), Lemon Ironbark (Eucalyptus staigeriana), Lemon mint,
Lemon Myrtle (Backhousia citriodora), Lemon Thyme, Lemon verbena
(Lippia citriodora), Licorice-adaptogen, Lime Flower, Limnophila
aromatica, Lingzhi, Linseed, Liquorice, Long pepper, Lovage
(Levisticum officinale), Luohanguo, Mace, Mahlab, Malabathrum,
Manchurian Thorn Tree (Aralia manchurica), Mandrake, Marjoram
(Origanum majorana), Marrubium vulgare, Marsh Labrador Tea,
Marshmallow, Mastic, Meadowsweet, Mei Yen, Melegueta pepper
(Aframomum melegueta), Mint, Milk thistle (Silybum), Bergamot
(Monarda didyma), Motherwort, Mountain Skullcap, Mullein (Verbascum
thapsus), Mustard, Mustard seed, Nashia inaguensis, Neem, Nepeta,
Nettle, Nigella sativa, Kolanji, Black caraway, Noni, Nutmeg, Mace,
Marijuana, Oenothera (Oenothera biennis), Olida (Eucalyptus olida),
Oregano (Origanum vulgare, O. heracleoticum), Orris root,
Osmorhiza, Olive Leaf (used in tea and as herbal supplement), Panax
quinquefolius, Pandan leaf, Paprika, Parsley (Petroselinum
crispum), Passion Flower, Patchouli, Pennyroyal, Pepper (black,
white, and green), Peppermint, Peppermint Gum (Eucalyptus dives),
Perilla, Plantain, Pomegranate, Ponch phoran, Poppy seed, Primrose
(Primula), candied flowers, dry tea mixes, Psyllium, Purslane,
Quassia, Quatre epices, Ramsons, Raspberry (leaves), Reishi,
Restharrow, Rhodiola rosea, Riberry (Syzygium luehmannii),
Rocket/Arugula, Roman chamomile, Rooibos, Rosehips, Rosemary
(Rosmarinus officinalis), Rowan Berries, Rue, Safflower, Saffron,
Sage (Salvia officinalis), Saigon Cinnamon, St John's Wort, Salad
Burnet (Sanguisorba minor or Poterium sanguisorba), Salvia, Sichuan
Pepper (Sansho), Sassafras, Savory (Satureja hortensis, S.
Montana), Schisandra (Schisandra chinensis), Scutellaria
costaricana, Senna (herb), Senna obtusifolia, Sesame seed, Sheep
Sorrel, Shepherd's Purse, Sialagogue, Siberian Chaga, Siberian
ginseng (Eleutherococcus senticosus), Siraitia grosvenorii
(luohanguo), Skullcap, Sloe Berries, Smudge Stick, Sonchus, Sorrel
(Rumex spp.), Southernwood, Spearmint, Speedwell, Squill, Star
anise, Stevia, Strawberry Leaves, Suma (Pfaffia paniculata), Sumac,
Summer savory, Sutherlandia frutescens, Sweet grass, Sweet cicely
(Myrrhis odorata), Sweet woodruff, Szechuan pepper (Xanthoxylum
piperitum), Tacamahac, Tamarind, Tandoori masala, Tansy, Tarragon
(Artemisia dracunculus), Tea, Teucrium polium, That basil, Thistle,
Thyme, Toor Dall, Tormentil, Tribulus terrestris, Tulsi (Ocimum
tenuiflorum), Turmeric (Curcuma longa), Uva Ursi also known as
Bearberry, Vanilla (Vanilla planifolia), Vasaka, Vervain, Vetiver,
Vietnamese Coriander (Persicaria odorata), Wasabi (Wasabia
japonica), Watercress, Wattleseed, Wild ginger, Wild Lettuce, Wild
thyme, Winter savory, Witch Hazel, Wolfberry, Wood Avens, Wood
Betony, Woodruff, Wormwood, Yarrow, Yerba Buena, Yohimbe, Za'atar,
Zedoary Root, or derivations thereof in aqueous or semi-aqueous
solution.
[0085] In an embodiment, myceliation of green coffee bean can
result in a decrease of compounds that is responsible for bitter
taste of coffee and caffeine content, thus coffee brewed from
roasted, myceliated coffee beans can eventually offers a smooth
drink of coffee with low caffeine content.
[0086] In an embodiment, extract obtained from agricultural
substrates myceliated by inoculating with pure culture of Ganoderma
lucidum, Pleurotus spp., Cordyceps sinensis can show enhanced
color, texture, flavor and smell.
[0087] In an embodiment, extract obtained from myceliated
substrates inoculated with pure cultures of fungal strains can be
selected from one or more of A. blazei, C. sinensis, G. lucidum, H.
erinaceus, G. frondosa, P. eryngii, P. ostreatus, P.
citrinopileatus, P. djamor, T versicolor, L. edodes, F. velutipes,
V. volvacea, H. marmoreus, P. nameko, T melanosporum, M hortensis,
P. umbellatus, and T. fuciformis, and can contain high level of
nucleic acids including RNA. The level of RNA content can be
greatly reduced by subjecting myceliated substrate to heat
treatment at a specific temperature for a predetermined period of
time followed by cooling at room temperature.
[0088] In an embodiment, extract can be formulated into a
nutraceutical composition comprising of a capsule, pill, tablet,
syrup, drink mix, fluid, lozenge, powder and combinations thereof,
by following the techniques known in the art.
[0089] In another embodiment, nutraceutical composition can further
comprise at least one pharmaceutically acceptable excipient
selected from the group comprising binder, coating substance,
disintegrant, filler, flavor, color, lubricant, glidant, sorbent,
preservative and sweetener.
[0090] In another embodiment, nutraceutical composition can further
comprise a dietary supplement selected from the group comprising
proteins, vitamins, minerals, amino acids, dietary fiber,
microelements and combinations thereof.
[0091] In an embodiment, extract and nutraceutical composition can
be produced using solid state or liquid state fermentation using
small batch mode or continuous mode of fermentation methods by
employing small scale container to large scale, commercial
fermenting vessel.
[0092] In an embodiment, extract comprises of bioactive compounds
such as polysaccharides, glucan, sterols, fatty acids, phenol,
triterpenes, proteins, amino acids, glycoproteins, vitamins,
mineral salts and its combinations thereof.
[0093] In an embodiment, extract comprises of health benefit
properties such as anti-aging, anti-tumor, anti-bacterial,
anti-fungal, anti-viral activity, cholesterol regulation,
neuroprotection, anti-oxidant, immunomodulation and
hepatoprotective properties.
[0094] While the preferred embodiments of the invention have been
described above, it will be recognized and understood that various
modifications may be made therein, and the appended claims are
intended to cover all such modifications which may fall within the
spirit and scope of the invention.
[0095] In the following, the present invention is described in more
detail by way of examples. However, these examples are not intended
to limit the scope of protection of the present invention in any
way.
EXAMPLES
Example 1
Decreasing Ribonucleic Acid Levels in Myceliated Substrate
[0096] Agricultural substrates completely myceliated by inoculating
with pure cultures of fungal strains selected from A. blazei, C.
sinensis, G. lucidum, H. erinaceus, G. frondosa, P. eryngii, P.
ostreatus, P. citrinopileatus, P. djamor, T. versicolor, L. edodes,
F. velutipes, V. volvacea, H. marmoreus, P. nameko, T.
melanosporum, M. hortensis, P. umbellatus, and T. fuciformis are
subjected to heat treatment prior to harvest for 1 min to 2 hours
at 145 degree F. to 195 degree F. followed by recovery at room
temperature for 45 min to 48 hours shows remarkable decrease in RNA
levels and are formulated into different nutraceutical
compositions.
Example 2
Extraction
[0097] 100 gram to 1000 grams of myceliated agricultural substrate
is extracted using 4 liters to 10 liters of boiling water and added
with 1 gram to 100 grams of citric acid and ascorbic acid and left
for 1 to 4 hours. The extract is filtered through a stainless steel
colander for separating a liquid extract, which is further
concentrated into 1/2 to 1/8 of the original volume by boiling and
stirring. To every 40 ml to 60 ml of hot liquid extract, 1 ml to 20
ml citrus essential oil and 100 grams to 200 grams dry honey are
added with vigorous stirring at 145 degree F. to 165 degree F. for
10 min to 45 min, then cooled down to 100 degree F. to 125 degree
F.
[0098] Honey formulations prepared from myceliated substrate in
accordance with the present invention are stirred for 10 min to 90
min at 100 degree F. to 125 degree F. and bottled for further use
or sale or storage.
Example 3
Nutraceutical Compositions
[0099] Raw flax seed and chia seed myceliated with Tremella
fuciformus, Pholiota nameko, Flammulina velutipes, and Ganoderma
lucidum, produces extraordinarily slippery jelly like product which
is filtered, purified and formulated as a nutraceutical composition
with clinical application including health beneficial properties
such as digestive enhancement and as a lubricant for mucus
membranes.
[0100] Extracts of fungi used herein are known to be of low
glycemic index which adds health-care benefits to the honey
formulations. This low-glyemic effect is enhanced by manipulating
the honey formulations into pills and or capsules, which take
longer to metabolize than the honey formulations by themselves.
Honey formulations are diluted at 100 degrees F. to 145 degrees F.
with 1 part honey formulations mixed with 2 to 10 parts purified
cellulose powder and/or 2 to 10 parts raw mushroom powder (w/w), as
filler, followed by mixing and pressing the mix into molds at 20
lb/in 2 to 200 lb/in 2 pressure at 70 degrees F. to 100 degrees F.
in order to make pills or capsules, which are then consumed in
order to enjoy the related health-care benefits of this
invention.
[0101] Similarly, suppositories, which also take longer to
metabolize than honey formulations themselves, are made as follows:
Mixing 1 to 10 parts of honey with 10 to 1000 parts of coconut
butter modified with 1 to 10 parts olive oil followed by heating
the mixture at 145 degrees F. to 175 degrees F. After vigorous
mixing, formulation is brought back to room temperature to cool,
and formulated into 1 inch to 2 inch long cylinders of 1/2 in
diameter and packed as nutraceutical tablets.
* * * * *