CN111620959A - Preparation method and application of ganoderma lucidum mycelium polysaccharide - Google Patents
Preparation method and application of ganoderma lucidum mycelium polysaccharide Download PDFInfo
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- CN111620959A CN111620959A CN202010514167.5A CN202010514167A CN111620959A CN 111620959 A CN111620959 A CN 111620959A CN 202010514167 A CN202010514167 A CN 202010514167A CN 111620959 A CN111620959 A CN 111620959A
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- ganoderma lucidum
- polysaccharide
- lucidum mycelium
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- mycelia
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- 229920001282 polysaccharide Polymers 0.000 title claims abstract description 120
- 239000005017 polysaccharide Substances 0.000 title claims abstract description 120
- 150000004676 glycans Chemical class 0.000 title claims abstract description 119
- 240000008397 Ganoderma lucidum Species 0.000 title claims abstract description 115
- 235000001637 Ganoderma lucidum Nutrition 0.000 title claims abstract description 115
- 238000002360 preparation method Methods 0.000 title claims abstract description 25
- 238000000605 extraction Methods 0.000 claims abstract description 61
- 238000005238 degreasing Methods 0.000 claims abstract description 15
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 13
- 239000003814 drug Substances 0.000 claims abstract description 6
- 229940079593 drug Drugs 0.000 claims abstract description 4
- 239000000843 powder Substances 0.000 claims description 32
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 28
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 24
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 claims description 20
- 239000003960 organic solvent Substances 0.000 claims description 20
- 108090000790 Enzymes Proteins 0.000 claims description 19
- 102000004190 Enzymes Human genes 0.000 claims description 19
- 229940088598 enzyme Drugs 0.000 claims description 19
- ARXJGSRGQADJSQ-UHFFFAOYSA-N 1-methoxypropan-2-ol Chemical compound COCC(C)O ARXJGSRGQADJSQ-UHFFFAOYSA-N 0.000 claims description 18
- 239000007788 liquid Substances 0.000 claims description 17
- 238000000034 method Methods 0.000 claims description 14
- 238000001694 spray drying Methods 0.000 claims description 14
- 238000003756 stirring Methods 0.000 claims description 14
- 108010059892 Cellulase Proteins 0.000 claims description 9
- 229940106157 cellulase Drugs 0.000 claims description 9
- 229940059442 hemicellulase Drugs 0.000 claims description 9
- 108010002430 hemicellulase Proteins 0.000 claims description 9
- 108010059345 keratinase Proteins 0.000 claims description 9
- 238000000926 separation method Methods 0.000 claims description 9
- 238000010438 heat treatment Methods 0.000 claims description 8
- 238000001816 cooling Methods 0.000 claims description 7
- 239000008367 deionised water Substances 0.000 claims description 7
- 229910021641 deionized water Inorganic materials 0.000 claims description 7
- 238000001035 drying Methods 0.000 claims description 7
- 238000001914 filtration Methods 0.000 claims description 7
- 239000002244 precipitate Substances 0.000 claims description 7
- 238000004321 preservation Methods 0.000 claims description 7
- 239000006228 supernatant Substances 0.000 claims description 7
- 230000009467 reduction Effects 0.000 claims description 5
- 206010028980 Neoplasm Diseases 0.000 claims description 4
- 239000000047 product Substances 0.000 claims description 4
- 230000032683 aging Effects 0.000 claims description 3
- 230000036772 blood pressure Effects 0.000 claims description 3
- 230000002526 effect on cardiovascular system Effects 0.000 claims description 3
- 230000036541 health Effects 0.000 claims description 3
- 230000007365 immunoregulation Effects 0.000 claims description 3
- 239000000203 mixture Substances 0.000 claims description 3
- 208000024172 Cardiovascular disease Diseases 0.000 claims description 2
- 206010011224 Cough Diseases 0.000 claims description 2
- 206010020751 Hypersensitivity Diseases 0.000 claims description 2
- 208000026935 allergic disease Diseases 0.000 claims description 2
- 230000007815 allergy Effects 0.000 claims description 2
- 208000026106 cerebrovascular disease Diseases 0.000 claims description 2
- 238000002156 mixing Methods 0.000 claims description 2
- 241000222336 Ganoderma Species 0.000 abstract description 16
- 239000002994 raw material Substances 0.000 abstract description 10
- 230000008901 benefit Effects 0.000 abstract description 5
- 230000000694 effects Effects 0.000 abstract description 5
- 239000002699 waste material Substances 0.000 abstract description 5
- 238000004519 manufacturing process Methods 0.000 abstract description 2
- 230000001376 precipitating effect Effects 0.000 abstract 1
- 230000000052 comparative effect Effects 0.000 description 13
- 238000009776 industrial production Methods 0.000 description 4
- 230000008569 process Effects 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- 150000002772 monosaccharides Chemical group 0.000 description 3
- 108090000623 proteins and genes Proteins 0.000 description 3
- 102000004169 proteins and genes Human genes 0.000 description 3
- SHZGCJCMOBCMKK-UHFFFAOYSA-N D-mannomethylose Natural products CC1OC(O)C(O)C(O)C1O SHZGCJCMOBCMKK-UHFFFAOYSA-N 0.000 description 2
- SRBFZHDQGSBBOR-IOVATXLUSA-N D-xylopyranose Chemical compound O[C@@H]1COC(O)[C@H](O)[C@H]1O SRBFZHDQGSBBOR-IOVATXLUSA-N 0.000 description 2
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 2
- 238000002835 absorbance Methods 0.000 description 2
- PYMYPHUHKUWMLA-UHFFFAOYSA-N arabinose Natural products OCC(O)C(O)C(O)C=O PYMYPHUHKUWMLA-UHFFFAOYSA-N 0.000 description 2
- SRBFZHDQGSBBOR-UHFFFAOYSA-N beta-D-Pyranose-Lyxose Natural products OC1COC(O)C(O)C1O SRBFZHDQGSBBOR-UHFFFAOYSA-N 0.000 description 2
- 235000019441 ethanol Nutrition 0.000 description 2
- 239000008103 glucose Substances 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 235000015097 nutrients Nutrition 0.000 description 2
- 238000001556 precipitation Methods 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- OCKGFTQIICXDQW-ZEQRLZLVSA-N 5-[(1r)-1-hydroxy-2-[4-[(2r)-2-hydroxy-2-(4-methyl-1-oxo-3h-2-benzofuran-5-yl)ethyl]piperazin-1-yl]ethyl]-4-methyl-3h-2-benzofuran-1-one Chemical compound C1=C2C(=O)OCC2=C(C)C([C@@H](O)CN2CCN(CC2)C[C@H](O)C2=CC=C3C(=O)OCC3=C2C)=C1 OCKGFTQIICXDQW-ZEQRLZLVSA-N 0.000 description 1
- 206010002198 Anaphylactic reaction Diseases 0.000 description 1
- WQZGKKKJIJFFOK-QTVWNMPRSA-N D-mannopyranose Chemical compound OC[C@H]1OC(O)[C@@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-QTVWNMPRSA-N 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- -1 Example 2A polysaccharide Chemical class 0.000 description 1
- PNNNRSAQSRJVSB-SLPGGIOYSA-N Fucose Natural products C[C@H](O)[C@@H](O)[C@H](O)[C@H](O)C=O PNNNRSAQSRJVSB-SLPGGIOYSA-N 0.000 description 1
- SHZGCJCMOBCMKK-DHVFOXMCSA-N L-fucopyranose Chemical compound C[C@@H]1OC(O)[C@@H](O)[C@H](O)[C@@H]1O SHZGCJCMOBCMKK-DHVFOXMCSA-N 0.000 description 1
- SHZGCJCMOBCMKK-JFNONXLTSA-N L-rhamnopyranose Chemical compound C[C@@H]1OC(O)[C@H](O)[C@H](O)[C@H]1O SHZGCJCMOBCMKK-JFNONXLTSA-N 0.000 description 1
- PNNNRSAQSRJVSB-UHFFFAOYSA-N L-rhamnose Natural products CC(O)C(O)C(O)C(O)C=O PNNNRSAQSRJVSB-UHFFFAOYSA-N 0.000 description 1
- 241000219071 Malvaceae Species 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- WQZGKKKJIJFFOK-PHYPRBDBSA-N alpha-D-galactose Chemical compound OC[C@H]1O[C@H](O)[C@H](O)[C@@H](O)[C@H]1O WQZGKKKJIJFFOK-PHYPRBDBSA-N 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 208000003455 anaphylaxis Diseases 0.000 description 1
- PYMYPHUHKUWMLA-WDCZJNDASA-N arabinose Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)C=O PYMYPHUHKUWMLA-WDCZJNDASA-N 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 235000019658 bitter taste Nutrition 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 230000000295 complement effect Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000003544 deproteinization Effects 0.000 description 1
- 238000001784 detoxification Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- 238000012869 ethanol precipitation Methods 0.000 description 1
- 239000000835 fiber Substances 0.000 description 1
- 229930182830 galactose Natural products 0.000 description 1
- 230000035784 germination Effects 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 235000021049 nutrient content Nutrition 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- OQUKIQWCVTZJAF-UHFFFAOYSA-N phenol;sulfuric acid Chemical compound OS(O)(=O)=O.OC1=CC=CC=C1 OQUKIQWCVTZJAF-UHFFFAOYSA-N 0.000 description 1
- 230000001766 physiological effect Effects 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 230000002035 prolonged effect Effects 0.000 description 1
- 230000004936 stimulating effect Effects 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 235000019640 taste Nutrition 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 150000003648 triterpenes Chemical class 0.000 description 1
- 238000002137 ultrasound extraction Methods 0.000 description 1
- 238000003809 water extraction Methods 0.000 description 1
Classifications
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0003—General processes for their isolation or fractionation, e.g. purification or extraction from biomass
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/125—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives containing carbohydrate syrups; containing sugars; containing sugar alcohols; containing starch hydrolysates
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P11/00—Drugs for disorders of the respiratory system
- A61P11/14—Antitussive agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/02—Immunomodulators
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/08—Antiallergic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P39/00—General protective or antinoxious agents
- A61P39/06—Free radical scavengers or antioxidants
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
- A61P9/12—Antihypertensives
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/54—Improvements relating to the production of bulk chemicals using solvents, e.g. supercritical solvents or ionic liquids
Abstract
The invention belongs to plant extractionThe technical field, in particular to a ganoderma lucidum mycelium polysaccharide and a preparation method and application thereof. The invention adopts ganoderma lucidum mycelium containing rich ganoderma lucidum polysaccharide as extraction raw material, and the ganoderma lucidum mycelium is subjected to pretreatment, degreasing treatment and supercritical CO2Extracting, performing enzymolysis, precipitating with ethanol, etc. to obtain Ganoderma mycelia polysaccharide. The extraction rate of the ganoderma lucidum mycelium polysaccharide prepared by the preparation method of the ganoderma lucidum mycelium polysaccharide provided by the invention reaches more than 15.4%, and the purity of the ganoderma lucidum mycelium polysaccharide reaches more than 76.1%. The invention adopts supercritical CO2The extraction technology and the enzymolysis technology carry out polysaccharide extraction on the ganoderma lucidum mycelia, the extraction rate of the polysaccharide is improved, the waste of raw materials is avoided, the production cost is saved, the prepared ganoderma lucidum mycelia polysaccharide is applied to the preparation of health-care products and medicines, the effect is good, and the economic benefit and the concordant benefit are obvious.
Description
Technical Field
The invention belongs to the technical field of plant extraction, and particularly relates to ganoderma lucidum mycelium polysaccharide and a preparation method and application thereof.
Background
The ganoderma lucidum mycelium is hypha formed by germination of ganoderma lucidum spores, more than half of nutrient substances required by ganoderma lucidum sporocarp are derived from nutrients stored by weight of the hypha, so that the nutrient content of the mycelium and the content of ganoderma lucidum mycelium polysaccharide are generally many times higher than those of the ganoderma lucidum sporocarp and ganoderma lucidum spore powder.
The ganoderma lucidum mycelium polysaccharide is composed of three strands of monosaccharide chains, the configuration of the ganoderma lucidum mycelium polysaccharide is similar to that of DNA and RNA, the ganoderma lucidum mycelium polysaccharide is a spiral three-dimensional structure, and the spiral layers are mainly fixed by hydrogen bonds. The ganoderma lucidum mycelium polysaccharide has obvious differences in certain physicochemical properties such as monosaccharide composition, glycosidic bond configuration, molecular weight, optical rotation, solubility and the like. Most of more than 200 kinds of extracted ganoderma lucidum mycelium polysaccharides are heteropolysaccharides, namely, the polysaccharides contain other monosaccharides such as galactose, mannose, arabinose, xylose, fucose and rhamnose besides glucose; due to different sources, the molecular weight can vary from tens of thousands to hundreds of thousands, most of them are accompanied by branches, and part of the polysaccharide also contains peptide chains.
Ganoderan is one of the most effective components in ganoderma, and has the characteristics of stimulating host nonspecific resistance, immunospecific reaction and inhibiting tumor physiological activity. In addition, the ganoderma lucidum polysaccharide has good effects in the aspects of immunoregulation, tumor resistance, aging resistance, cardiovascular and cerebrovascular protection, liver protection, detoxification, blood pressure reduction, anaphylactic reaction reduction and the like. The ganoderma lucidum polysaccharide is the soul of ganoderma lucidum function, the content of ganoderma lucidum polysaccharide determines the efficacy of the product, and the ganoderma lucidum polysaccharide is the main component for treating various diseases.
Patent text with application number ZL201910800585.8 discloses a ganoderma lucidum polysaccharide extraction technology, and the extraction technology comprises the steps of raw material pretreatment, ganoderma lucidum crude polysaccharide extraction, ganoderma lucidum polysaccharide deproteinization, ganoderma lucidum extract purification, alcohol precipitation extract and the like, wherein the ganoderma lucidum crude polysaccharide extraction adopts hot water extraction, is assisted by ultrasonic extraction, and is further purified through the steps of enzymolysis of protein and alcohol precipitation, and finally the ganoderma lucidum polysaccharide is obtained. Although the extraction method can obtain a certain amount of ganoderan, the purity of ganoderan is not high, and the ganoderma is used as the extraction raw material, which causes a great amount of raw material waste in the extraction process, has high extraction cost, and is not suitable for industrial production.
In conclusion, the technical problems of low purity of ganoderma lucidum polysaccharide, serious waste of raw materials, unsuitability for industrial production and the like generally exist in the prior art.
Disclosure of Invention
In order to overcome the defects of the prior art, the invention aims to provide a preparation method and application of ganoderma lucidum mycelium polysaccharide. The ganoderma lucidum mycelium polysaccharide provided by the invention has high purity and good effect, directly adopts ganoderma lucidum mycelium with rich polysaccharide content as an extraction raw material, avoids the waste of the raw material, saves the cost, and is beneficial to realizing industrial production.
In order to achieve the purpose, the technical scheme of the invention is as follows:
a preparation method of ganoderma lucidum mycelium polysaccharide specifically comprises the following steps:
s1, putting the dried ganoderma lucidum mycelia into a grinder to be ground to 8-10 mu m, then adding an organic solvent to carry out degreasing treatment, carrying out centrifugal separation, taking the lower-layer precipitate, and drying at 50-70 ℃ to obtain ganoderma lucidum mycelia coarse powder;
s2, adding 10-20 times of deionized water into the ganoderma lucidum mycelium coarse powder prepared in the step S1, and then performing supercritical CO2Extracting, filtering to obtain water extract;
s3, taking the water extract prepared in the step S2, adding complex enzyme, and carrying out heat preservation enzymolysis under the stirring state of 400-500 rpm for 3-5 hours to prepare enzymolysis liquid;
s4, heating the enzymolysis liquid prepared in the step S3 to 100-120 ℃, preserving heat for 1 hour, naturally cooling to 20 ℃, and then performing spray drying to prepare polysaccharide coarse powder;
s5, adding 8-10 times of absolute ethyl alcohol into the polysaccharide coarse powder prepared in the step S4, stirring for 1-1.5 hours under the condition of 600-800 rpm, performing centrifugal separation, removing supernatant, and performing spray drying to obtain the polysaccharide coarse powder.
Further, the adding amount of the organic solvent in the step S1 is 8-10 times of the mass of the ganoderma lucidum mycelia; the organic solvent is a mixture of propylene glycol methyl ether, acetone and dimethyl sulfoxide; the degreasing time is 1-3 h.
Furthermore, the mixing mass ratio of the propylene glycol methyl ether, the acetone and the dimethyl sulfoxide is 2-3: 4-7: 1-2.
Further, supercritical CO in step S22The extraction conditions are as follows: the pressure is 30-35 Mpa, the extraction temperature is 50-60 ℃, and the extraction time is 4-5 h.
Further, the complex enzyme in the step S3 is composed of cellulase, keratinase and hemicellulase according to a mass ratio of 5-7: 1-3: 3-5.
Further, the volume fraction of the absolute ethyl alcohol in the step S5 is 95%.
The invention also provides the application of the ganoderma lucidum mycelium polysaccharide prepared by the preparation method of the ganoderma lucidum mycelium polysaccharide in health care products and medicines.
Furthermore, the medicine comprises tablets and oral liquid for treating immunoregulation, tumor resistance, aging resistance, cardiovascular and cerebrovascular diseases, blood pressure reduction, cough relieving and allergy resistance.
The invention adopts the ganoderma lucidum mycelia as the extraction raw material of the ganoderma lucidum mycelia polysaccharide, effectively solves the problem of bitter taste of the fruiting body polysaccharide, and the prepared ganoderma lucidum mycelia polysaccharide has better taste when being applied to oral liquid. In addition, the fiber content of the ganoderma lucidum mycelia is low, the content of the basswood polysaccharide is high, and the content of the triterpene component in the ganoderma lucidum mycelia is low, so that the extraction rate of the ganoderma lucidum mycelia polysaccharide is high, the extraction of the ganoderma lucidum mycelia polysaccharide by adopting the ganoderma lucidum mycelia can effectively improve the extraction rate of the polysaccharide, the process steps of later-stage polysaccharide purification are simplified, and the comprehensive utilization effect is good.
The preparation method of the ganoderma lucidum mycelium polysaccharide provided by the invention adopts organic solvent for treatment, and can effectively remove ganoderma lucidumThe esters contained in the ganoderma lucidum mycelia lay the foundation for the efficient extraction of ganoderma lucidum mycelia polysaccharide. Supercritical CO2The extraction technique is to use supercritical CO2Dissolving Ganoderma mycelia polysaccharide by supercritical CO under pressure and temperature2The ganoderma lucidum mycelium polysaccharide is extracted under the supercritical state. The complex enzyme consisting of cellulase, keratinase and hemicellulase is adopted to treat the water extract, so that protein, amino acid and nucleic acid in the water extract can be effectively removed, the purity of ganoderma lucidum mycelium polysaccharide is improved, and the enzyme can be effectively inactivated by heating to 100-120 ℃ after enzymolysis and preserving heat for 1 h. And finally, removing protein and small molecular impurities by adopting an ethanol precipitation method, and further improving the purity of the ganoderma lucidum mycelium polysaccharide.
Compared with the prior art, the invention has the following advantages;
(1) the extraction rate of the ganoderma lucidum mycelium polysaccharide prepared by the preparation method of the ganoderma lucidum mycelium polysaccharide provided by the invention reaches more than 15.4%, and the purity of the ganoderma lucidum mycelium polysaccharide reaches more than 76.1%;
(2) the invention adopts supercritical CO2The extraction technology and the enzymolysis technology are complementary to each other for polysaccharide extraction of the ganoderma lucidum mycelia, so that the extraction rate of the polysaccharide is improved, the waste of raw materials is avoided, and the production cost is saved;
(3) the preparation method of the ganoderma lucidum mycelium polysaccharide provided by the invention is simple in process, controllable in conditions and easy to realize industrial production; the prepared ganoderma lucidum mycelium polysaccharide is applied to the preparation of health care products and medicines, has good efficacy and obvious economic benefit and concordant benefit.
Detailed Description
The present invention will be further described below by way of specific embodiments, but the present invention is not limited to only the following examples. Various modifications can be made by those skilled in the art based on the basic idea of the invention, but it is within the scope of the invention as long as it does not depart from the basic idea of the invention.
Example 1A Ganoderma mycelia polysaccharide
The preparation method of the ganoderma lucidum mycelium polysaccharide comprises the following steps:
s1, putting the dried ganoderma lucidum mycelia into a grinder to be ground to 8 microns, then adding 8 times of organic solvent to carry out degreasing treatment, wherein the organic solvent consists of propylene glycol monomethyl ether, acetone and dimethyl sulfoxide according to the mass ratio of 2:4:1, the degreasing treatment time is 1h, carrying out centrifugal separation, taking the lower layer precipitate, and drying at 50 ℃ to obtain ganoderma lucidum mycelia coarse powder;
s2, adding 10 times of deionized water into the ganoderma lucidum mycelium coarse powder prepared in the step S1, and then performing supercritical CO2Extracting with supercritical CO2The extraction conditions are as follows: the pressure is 30Mpa, the extraction temperature is 50 ℃, and the extraction time is 4 h; filtering to obtain water extractive solution;
s3, taking the water extract prepared in the step S2, adding a complex enzyme, wherein the complex enzyme consists of cellulase, keratinase and hemicellulase according to a mass ratio of 5:1:3, and performing heat preservation enzymolysis under a stirring state of 400 revolutions per minute for 3 hours to prepare an enzymolysis liquid;
s4, heating the enzymolysis liquid prepared in the step S3 to 100 ℃, preserving heat for 1 hour, naturally cooling to 20 ℃, and then performing spray drying to prepare polysaccharide coarse powder;
s5, adding 8 times of 95% absolute ethyl alcohol by volume fraction into the polysaccharide coarse powder prepared in the step S4, stirring for 1 hour under the condition of 600 revolutions per minute, centrifugally separating, removing supernatant, and performing spray drying to obtain the polysaccharide.
Example 2A polysaccharide of Ganoderma mycelia
The preparation method of the ganoderma lucidum mycelium polysaccharide comprises the following steps:
s1, putting the dried ganoderma lucidum mycelia into a grinder to be ground to 8.5 microns, then adding 8.5 times of organic solvent for degreasing treatment, wherein the organic solvent consists of propylene glycol methyl ether, acetone and dimethyl sulfoxide according to the mass ratio of 3:5:2, the degreasing treatment time is 1.5h, carrying out centrifugal separation, taking the lower layer precipitate, and drying at 55 ℃ to obtain ganoderma lucidum mycelia coarse powder;
s2, adding 14 times of deionized water into the ganoderma lucidum mycelium coarse powder prepared in the step S1, and then performing supercritical CO2Extracting with supercritical CO2Conditions of extractionComprises the following steps: the pressure is 31Mpa, the extraction temperature is 53 ℃, and the extraction time is 4.2 h; filtering to obtain water extractive solution;
s3, taking the water extract prepared in the step S2, adding a complex enzyme, wherein the complex enzyme consists of cellulase, keratinase and hemicellulase according to the mass ratio of 6:2:3, and performing heat preservation enzymolysis under the stirring state of 420 revolutions per minute for 3.5 hours to prepare an enzymolysis liquid;
s4, heating the enzymolysis liquid prepared in the step S3 to 105 ℃, preserving heat for 1 hour, naturally cooling to 20 ℃, and then performing spray drying to prepare polysaccharide coarse powder;
s5, adding 8.5 times of 95% absolute ethyl alcohol by volume fraction into the polysaccharide coarse powder prepared in the step S4, stirring for 1.2 hours under the condition of 650 revolutions per minute, centrifugally separating, removing supernatant, and performing spray drying to obtain the polysaccharide.
Example 3A Ganoderma mycelia polysaccharide
The preparation method of the ganoderma lucidum mycelium polysaccharide comprises the following steps:
s1, putting the dried ganoderma lucidum mycelia into a grinder to be ground to 9 microns, then adding 9 times of organic solvent for degreasing treatment, wherein the organic solvent comprises propylene glycol methyl ether, acetone and dimethyl sulfoxide according to the mass ratio of 2:6:1, the degreasing treatment time is 2 hours, carrying out centrifugal separation, taking the lower layer precipitate, and drying at 58 ℃ to obtain ganoderma lucidum mycelia coarse powder;
s2, adding 16 times of deionized water into the ganoderma lucidum mycelium coarse powder prepared in the step S1, and then performing supercritical CO2Extracting with supercritical CO2The extraction conditions are as follows: the pressure is 33Mpa, the extraction temperature is 55 ℃, and the extraction time is 4.5 h; filtering to obtain water extractive solution;
s3, taking the water extract prepared in the step S2, adding a complex enzyme, wherein the complex enzyme consists of cellulase, keratinase and hemicellulase according to a mass ratio of 7:2:4, and performing heat preservation enzymolysis under a stirring state of 450 revolutions per minute for 4 hours to prepare an enzymolysis liquid;
s4, heating the enzymolysis liquid prepared in the step S3 to 110 ℃, preserving heat for 1 hour, naturally cooling to 20 ℃, and then performing spray drying to prepare polysaccharide coarse powder;
s5, adding 9 times of 95% absolute ethyl alcohol by volume fraction into the polysaccharide coarse powder prepared in the step S4, stirring for 1.3 hours under the condition of 700 revolutions per minute, centrifugally separating, removing supernatant, and performing spray drying to obtain the polysaccharide.
Example 4A polysaccharide of Ganoderma mycelia
The preparation method of the ganoderma lucidum mycelium polysaccharide comprises the following steps:
s1, putting the dried ganoderma lucidum mycelia into a grinder to be ground to 9.5 mu m, then adding 9.5 times of organic solvent for degreasing treatment, wherein the organic solvent consists of propylene glycol methyl ether, acetone and dimethyl sulfoxide according to the mass ratio of 3:7:2, the degreasing treatment time is 2.5h, carrying out centrifugal separation, taking the lower layer precipitate, and drying at 65 ℃ to obtain ganoderma lucidum mycelia coarse powder;
s2, adding 18 times of deionized water into the ganoderma lucidum mycelium coarse powder prepared in the step S1, and then performing supercritical CO2Extracting with supercritical CO2The extraction conditions are as follows: the pressure is 34Mpa, the extraction temperature is 58 ℃, and the extraction time is 4.7 h; filtering to obtain water extractive solution;
s3, taking the water extract prepared in the step S2, adding a complex enzyme, wherein the complex enzyme consists of cellulase, keratinase and hemicellulase according to the mass ratio of 7:3:5, and performing heat preservation enzymolysis under the stirring state of 470 revolutions per minute for 4.5 hours to prepare an enzymolysis liquid;
s4, heating the enzymolysis liquid prepared in the step S3 to 115 ℃, preserving heat for 1 hour, naturally cooling to 20 ℃, and then performing spray drying to prepare polysaccharide coarse powder;
s5, adding 9.5 times of 95% absolute ethyl alcohol by volume fraction into the polysaccharide coarse powder prepared in the step S4, stirring for 1.4 hours at 750 revolutions per minute, centrifugally separating, removing supernatant, and spray drying to obtain the polysaccharide.
Example 5A polysaccharide of Ganoderma mycelia
The preparation method of the ganoderma lucidum mycelium polysaccharide comprises the following steps:
s1, putting the dried ganoderma lucidum mycelia into a grinder to be ground to 10 microns, then adding 10 times of organic solvent to carry out degreasing treatment, wherein the organic solvent consists of propylene glycol monomethyl ether, acetone and dimethyl sulfoxide according to the mass ratio of 3:6:1, the degreasing treatment time is 3 hours, carrying out centrifugal separation, taking the lower layer precipitate, and drying at 70 ℃ to obtain ganoderma lucidum mycelia coarse powder;
s2, adding 20 times of deionized water into the ganoderma lucidum mycelium coarse powder prepared in the step S1, and then performing supercritical CO2Extracting with supercritical CO2The extraction conditions are as follows: extracting under 35Mpa at 50 deg.C for 5 hr; filtering to obtain water extractive solution;
s3, taking the water extract prepared in the step S2, adding a complex enzyme, wherein the complex enzyme consists of cellulase, keratinase and hemicellulase according to the mass ratio of 6:2:5, and performing heat preservation enzymolysis under the stirring state of 500 revolutions per minute for 5 hours to prepare an enzymolysis liquid;
s4, heating the enzymolysis liquid prepared in the step S3 to 120 ℃, preserving heat for 1 hour, naturally cooling to 20 ℃, and then performing spray drying to prepare polysaccharide coarse powder;
s5, adding 10 times of 95% absolute ethyl alcohol by volume fraction into the polysaccharide coarse powder prepared in the step S4, stirring for 1.5 hours under the condition of 800 revolutions per minute, centrifugally separating, removing supernatant, and performing spray drying to obtain the polysaccharide.
Comparative example 1A Ganoderma mycelia polysaccharide
The preparation method of the ganoderma lucidum mycelium polysaccharide is similar to that of example 3.
The difference between this comparative example and example 3 is: the organic solvent in step S1 in this comparative example was composed of propylene glycol methyl ether, acetone, and dimethyl sulfoxide in a mass ratio of 1:1: 1.
Comparative example 2A Ganoderma mycelia polysaccharide
The preparation method of the ganoderma lucidum mycelium polysaccharide is similar to that of example 3.
The difference between this comparative example and example 3 is: supercritical CO in step S2 of this comparative example2The extraction conditions are as follows: the pressure is 25Mpa, the extraction temperature is 80 ℃, and the extraction time is 6 h.
Comparative example 3A Ganoderma mycelia polysaccharide
The preparation method of the ganoderma lucidum mycelium polysaccharide is similar to that of example 3.
The difference between this comparative example and example 3 is: the complex enzyme in step S3 in this comparative example consists of cellulase, keratinase and hemicellulase in a mass ratio of 1:1: 1.
Test example, measurement of polysaccharide content in Ganoderma mycelia
Test samples: ganoderma lucidum mycelia polysaccharides obtained in examples 1 to 5 and comparative examples 1 to 3;
the test method comprises the following steps: a standard curve is drawn by adopting a phenol-sulfuric acid method and taking the glucose content c as an abscissa (mu g) and the absorbance A as an ordinate, so that a linear equation A is 0.0067c +0.0005, and the correlation coefficient is 0.9992. Preparing ganoderma lucidum mycelium polysaccharide into polysaccharide solution with the molar concentration of 0.1mol/L, performing color development by the same method, measuring absorbance A, and substituting into an equation to calculate the content of the ganoderma lucidum mycelium polysaccharide; the extraction rate of the ganoderma lucidum mycelium polysaccharide is the ratio of the content of the ganoderma lucidum mycelium polysaccharide to the content of ganoderma lucidum mycelium powder.
And (3) test results: the test results are shown in Table 1.
TABLE 1 measurement of polysaccharide content in mycelia of Ganoderma lucidum
As can be seen from Table 1, the process for extracting polysaccharides from ganoderma lucidum mycelia provided by the invention can realize that the extraction rate of the ganoderma lucidum mycelia polysaccharides reaches more than 15.4%, and the purity of the extracted ganoderma lucidum mycelia polysaccharides reaches more than 76.1%, so that the process has great progress compared with the prior art. The ganoderma lucidum mycelium polysaccharide prepared in the embodiment 3 has the highest extraction rate and the best purity, and is the best embodiment of the invention.
Compared with example 3, comparative example 1 changed the amount ratio of organic solvent used in the defatting process, but the obtained ganoderan had lower purity and lower extraction yield, which indicates that the ratio of organic solvent in the preparation method of ganoderan provided by the present invention is already lowThe optimal proportion is achieved, and the adverse effect of reducing the extraction rate can be caused by changing the dosage ratio. Comparative example 2 changed supercritical CO2The extraction conditions, such as reduced extraction pressure, increased extraction temperature, and prolonged extraction time, are obviously inferior to those of example 3 in the extraction yield and purity of the Ganoderma mycelia polysaccharides due to the reduction of supercritical CO2The pressure of extraction can reduce the content of Ganoderma mycelia polysaccharide in supercritical CO2The solubility in water, resulting in a decrease in extraction yield, and neither an increase in extraction temperature nor an extension in extraction time can compensate for this drawback. Comparative example 3 changes the dosage ratio of the complex enzyme in step S3, but the extraction rate of the ganoderma lucidum mycelia polysaccharide and the purity of the ganoderma lucidum mycelia polysaccharide are obviously inferior to those of example 3, which shows that the ratio of the components contained in the complex enzyme in the preparation method of the ganoderma lucidum mycelia polysaccharide provided by the invention has reached the optimal ratio.
The foregoing embodiments are merely illustrative of the principles and utilities of the present invention and are not intended to limit the invention. Those skilled in the art will recognize that changes may be made to the embodiments described above without departing from the spirit and scope of the invention. Therefore, it is intended that all equivalent modifications or changes which can be made by those skilled in the art without departing from the technical spirit of the present invention are covered by the claims of the present invention.
Claims (8)
1. The preparation method of the ganoderma lucidum mycelium polysaccharide is characterized by comprising the following steps:
s1, putting the dried ganoderma lucidum mycelia into a grinder to be ground to 8-10 mu m, then adding an organic solvent to carry out degreasing treatment, carrying out centrifugal separation, taking the lower-layer precipitate, and drying at 50-70 ℃ to obtain ganoderma lucidum mycelia coarse powder;
s2, adding 10-20 times of deionized water into the ganoderma lucidum mycelium coarse powder prepared in the step S1, and then performing supercritical CO2Extracting, filtering to obtain water extract;
s3, taking the water extract prepared in the step S2, adding complex enzyme, and carrying out heat preservation enzymolysis under the stirring state of 400-500 rpm for 3-5 hours to prepare enzymolysis liquid;
s4, heating the enzymolysis liquid prepared in the step S3 to 100-120 ℃, preserving heat for 1 hour, naturally cooling to 20 ℃, and then performing spray drying to prepare polysaccharide coarse powder;
s5, adding 8-10 times of absolute ethyl alcohol into the polysaccharide coarse powder prepared in the step S4, stirring for 1-1.5 hours under the condition of 600-800 rpm, performing centrifugal separation, removing supernatant, and performing spray drying to obtain the polysaccharide coarse powder.
2. The method for preparing ganoderma lucidum mycelium polysaccharide according to claim 1, wherein the amount of the organic solvent added in the step S1 is 8-10 times of the mass of ganoderma lucidum mycelium; the organic solvent is a mixture of propylene glycol methyl ether, acetone and dimethyl sulfoxide; the degreasing time is 1-3 h.
3. The method for preparing ganoderma lucidum mycelium polysaccharide according to claim 2, wherein the mixing mass ratio of propylene glycol methyl ether, acetone and dimethyl sulfoxide is 2-3: 4-7: 1-2.
4. The method for preparing the ganoderma lucidum mycelium polysaccharide according to claim 1, wherein supercritical CO is adopted in the step S22The extraction conditions are as follows: the pressure is 30-35 Mpa, the extraction temperature is 50-60 ℃, and the extraction time is 4-5 h.
5. The preparation method of the ganoderma lucidum mycelium polysaccharide according to claim 1, wherein the complex enzyme in the step S3 is composed of cellulase, keratinase and hemicellulase according to a mass ratio of 5-7: 1-3: 3-5.
6. The method for preparing ganoderma lucidum mycelium polysaccharide according to claim 1, wherein the volume fraction of the absolute ethyl alcohol in the step S5 is 95%.
7. Use of the ganoderma lucidum mycelia polysaccharide prepared by the method for preparing ganoderma lucidum mycelia polysaccharide as claimed in claim 1 to 6 in health products and medicines.
8. The use of the ganoderma lucidum mycelia polysaccharides according to claim 7, wherein the medicament comprises tablets and oral liquids for treating immunoregulation, tumor resistance, aging resistance, cardiovascular and cerebrovascular diseases, blood pressure reduction, cough relieving and allergy resisting.
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