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.

Description

RELATED APPLICATIONS

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

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

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.

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.

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.

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.

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-β-glucans and their protein complexes (e.g., xyloglucans, and acidic (β-glucan containing uronic acid) as well as dietary fiber, lectins, and triterpenoids, have been isolated from medicinal mushrooms.

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.

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.

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.

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.

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

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.

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.

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.

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

• • 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 a 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;
  • wherein the at least one small molecule compound enhances bioavailability, water solubility and passive absorption of the aqueous extract.

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:

• • optionally pre-processing an agricultural substrate depending on nature of the substrate;
  • 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;
  • enabling mycelial growth on the agricultural substrate till the mycelial growth reaches a desired stage to obtain a myceliated agricultural substrate;
  • obtaining a water-substrate mixture by boiling the myceliated agricultural substrate in water for a suitable duration of time and at a 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 nutraceutical or food composition using techniques known in the art;

wherein the at least one small molecule compound enhances bioavailability, water solubility and passive absorption of the aqueous extract.

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.

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.

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

FIG. 1 is a flowchart of a method for preparing an extract of myceliated agricultural substrates.

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

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.

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.

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.

FIG. 1 shows a flow chart of a method of preparation of an extract from myceliated agricultural substrates wherein,

• • step 10 provides an agricultural substrate;
  • step 12 optionally pre-processes the substrate based on nature of the substrate,
  • step 14 inoculates an agricultural substrate with pure culture of fungal strains selected from Basidiomycota and Ascomycota;
  • step 16 enables mycelium growth on the substrate under optimum growth conditions to obtain a myceliated agricultural substrate;
  • step 18 grows the mycelium on the myceliated agricultural substrate till the mycelial growth reaches a desired stage;
  • step 20 obtains a water substrate mixture by boiling the myceliated agricultural substrate with water;
  • step 22 separates aqueous and non-aqueous components from the water substrate mixture; and
  • step 24 adds at least one small molecule component to the aqueous component to form the aqueous extract of the myceliated substrate.

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,

• • step 30 provides an agricultural substrate,
  • step 32 optionally pre-processes the substrate based on nature of the substrate,
  • step 34 inoculates the substrate with pure culture of fungal strains selected from Tremella fuciformis and Hericium erinaceus,
  • step 36 enables mycelium growth on the substrate under optimum growth conditions till a desired stage is reached to obtain a myceliated substrate;
  • step 38 obtains a water substrate mixture by boiling the myceliated substrate with water;
  • step 40 separates aqueous and non-aqueous components from the water substrate mixture;
  • step 42 adding at least one small molecule compound to the aqueous extract; and
  • step 44 optionally formulates the aqueous extract into a nutraceutical or food composition using techniques known in the art.

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.

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.

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.

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.

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.

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.

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.

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.

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.

In a yet another embodiment, liquid media can be injected with sterile air to deliver oxygen to the liquid media.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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

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

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 ½ to ⅛ 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.

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

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.

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.

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 ½ in diameter and packed as nutraceutical tablets.

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.