CN116694702A - Product rich in beta-glucan and polypeptide as well as preparation method and application thereof - Google Patents
Product rich in beta-glucan and polypeptide as well as preparation method and application thereof Download PDFInfo
- Publication number
- CN116694702A CN116694702A CN202310702412.9A CN202310702412A CN116694702A CN 116694702 A CN116694702 A CN 116694702A CN 202310702412 A CN202310702412 A CN 202310702412A CN 116694702 A CN116694702 A CN 116694702A
- Authority
- CN
- China
- Prior art keywords
- glucan
- beta
- polypeptide
- product
- supernatant
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 229920002498 Beta-glucan Polymers 0.000 title claims abstract description 101
- FYGDTMLNYKFZSV-URKRLVJHSA-N (2s,3r,4s,5s,6r)-2-[(2r,4r,5r,6s)-4,5-dihydroxy-2-(hydroxymethyl)-6-[(2r,4r,5r,6s)-4,5,6-trihydroxy-2-(hydroxymethyl)oxan-3-yl]oxyoxan-3-yl]oxy-6-(hydroxymethyl)oxane-3,4,5-triol Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1[C@@H](CO)O[C@@H](OC2[C@H](O[C@H](O)[C@H](O)[C@H]2O)CO)[C@H](O)[C@H]1O FYGDTMLNYKFZSV-URKRLVJHSA-N 0.000 title claims abstract description 100
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 93
- 229920001184 polypeptide Polymers 0.000 title claims abstract description 91
- 102000004196 processed proteins & peptides Human genes 0.000 title claims abstract description 91
- 238000002360 preparation method Methods 0.000 title claims abstract description 18
- 239000000047 product Substances 0.000 claims abstract description 90
- 239000006228 supernatant Substances 0.000 claims abstract description 59
- 239000004365 Protease Substances 0.000 claims abstract description 47
- 241000222481 Schizophyllum commune Species 0.000 claims abstract description 41
- 102000004190 Enzymes Human genes 0.000 claims abstract description 35
- 108090000790 Enzymes Proteins 0.000 claims abstract description 35
- 235000007340 Hordeum vulgare Nutrition 0.000 claims abstract description 29
- 238000000855 fermentation Methods 0.000 claims abstract description 26
- 230000004151 fermentation Effects 0.000 claims abstract description 26
- 239000002244 precipitate Substances 0.000 claims abstract description 24
- 238000000605 extraction Methods 0.000 claims abstract description 23
- 235000007164 Oryza sativa Nutrition 0.000 claims abstract description 22
- 108091005804 Peptidases Proteins 0.000 claims abstract description 22
- 235000009566 rice Nutrition 0.000 claims abstract description 22
- 150000001875 compounds Chemical class 0.000 claims abstract description 19
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 13
- 239000000843 powder Substances 0.000 claims abstract description 11
- 238000001035 drying Methods 0.000 claims abstract description 9
- 239000002994 raw material Substances 0.000 claims abstract description 9
- 230000001105 regulatory effect Effects 0.000 claims abstract description 7
- 230000000415 inactivating effect Effects 0.000 claims abstract description 6
- 238000007873 sieving Methods 0.000 claims abstract description 6
- 230000001954 sterilising effect Effects 0.000 claims abstract description 6
- 240000007594 Oryza sativa Species 0.000 claims abstract 4
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 claims abstract 3
- 229940088598 enzyme Drugs 0.000 claims description 33
- 241000209219 Hordeum Species 0.000 claims description 27
- 108090000526 Papain Proteins 0.000 claims description 25
- 229940055729 papain Drugs 0.000 claims description 25
- 235000019834 papain Nutrition 0.000 claims description 25
- 239000000243 solution Substances 0.000 claims description 23
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 22
- 235000019419 proteases Nutrition 0.000 claims description 21
- 239000007788 liquid Substances 0.000 claims description 17
- 108010059892 Cellulase Proteins 0.000 claims description 13
- 108090000787 Subtilisin Proteins 0.000 claims description 13
- 102000004142 Trypsin Human genes 0.000 claims description 13
- 108090000631 Trypsin Proteins 0.000 claims description 13
- 229940106157 cellulase Drugs 0.000 claims description 13
- 239000012588 trypsin Substances 0.000 claims description 13
- 239000004382 Amylase Substances 0.000 claims description 12
- 102000013142 Amylases Human genes 0.000 claims description 12
- 108010065511 Amylases Proteins 0.000 claims description 12
- 235000019418 amylase Nutrition 0.000 claims description 12
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 9
- 239000007864 aqueous solution Substances 0.000 claims description 8
- 229920005654 Sephadex Polymers 0.000 claims description 7
- 239000012507 Sephadex™ Substances 0.000 claims description 7
- 238000004108 freeze drying Methods 0.000 claims description 6
- 239000012528 membrane Substances 0.000 claims description 6
- 238000000926 separation method Methods 0.000 claims description 6
- 239000004743 Polypropylene Substances 0.000 claims description 5
- 238000001914 filtration Methods 0.000 claims description 5
- 238000001471 micro-filtration Methods 0.000 claims description 5
- -1 polypropylene Polymers 0.000 claims description 5
- 229920001155 polypropylene Polymers 0.000 claims description 5
- 230000001376 precipitating effect Effects 0.000 claims description 5
- 238000000746 purification Methods 0.000 claims description 5
- 230000001580 bacterial effect Effects 0.000 claims description 4
- 239000002537 cosmetic Substances 0.000 claims description 4
- 235000013312 flour Nutrition 0.000 claims description 3
- 238000005227 gel permeation chromatography Methods 0.000 claims description 2
- 238000002604 ultrasonography Methods 0.000 claims 3
- 230000002255 enzymatic effect Effects 0.000 claims 1
- 239000000413 hydrolysate Substances 0.000 claims 1
- 238000005516 engineering process Methods 0.000 abstract description 3
- 238000006243 chemical reaction Methods 0.000 abstract description 2
- 240000005979 Hordeum vulgare Species 0.000 abstract 2
- 238000012360 testing method Methods 0.000 description 28
- 230000000052 comparative effect Effects 0.000 description 25
- 102000035195 Peptidases Human genes 0.000 description 19
- 241000209094 Oryza Species 0.000 description 18
- 241000222480 Schizophyllum Species 0.000 description 18
- 210000003491 skin Anatomy 0.000 description 16
- 238000000034 method Methods 0.000 description 15
- 210000004027 cell Anatomy 0.000 description 13
- 102000004169 proteins and genes Human genes 0.000 description 13
- 108090000623 proteins and genes Proteins 0.000 description 13
- 238000010586 diagram Methods 0.000 description 12
- 229920001282 polysaccharide Polymers 0.000 description 12
- 239000005017 polysaccharide Substances 0.000 description 12
- 150000004804 polysaccharides Chemical class 0.000 description 12
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 9
- 239000000126 substance Substances 0.000 description 9
- 235000016709 nutrition Nutrition 0.000 description 7
- 239000000523 sample Substances 0.000 description 7
- 210000002421 cell wall Anatomy 0.000 description 6
- 235000013339 cereals Nutrition 0.000 description 6
- 230000003834 intracellular effect Effects 0.000 description 6
- 239000013641 positive control Substances 0.000 description 6
- 241000233866 Fungi Species 0.000 description 5
- 210000000245 forearm Anatomy 0.000 description 5
- 230000008439 repair process Effects 0.000 description 5
- 150000001413 amino acids Chemical class 0.000 description 4
- 230000007071 enzymatic hydrolysis Effects 0.000 description 4
- 238000006047 enzymatic hydrolysis reaction Methods 0.000 description 4
- 238000005259 measurement Methods 0.000 description 4
- 230000003020 moisturizing effect Effects 0.000 description 4
- 230000035764 nutrition Effects 0.000 description 4
- 238000011218 seed culture Methods 0.000 description 4
- 230000008591 skin barrier function Effects 0.000 description 4
- 238000012258 culturing Methods 0.000 description 3
- 238000009630 liquid culture Methods 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 3
- 238000002156 mixing Methods 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 231100000241 scar Toxicity 0.000 description 3
- 239000008223 sterile water Substances 0.000 description 3
- 238000010998 test method Methods 0.000 description 3
- GVJHHUAWPYXKBD-UHFFFAOYSA-N (±)-α-Tocopherol Chemical compound OC1=C(C)C(C)=C2OC(CCCC(C)CCCC(C)CCCC(C)C)(C)CCC2=C1C GVJHHUAWPYXKBD-UHFFFAOYSA-N 0.000 description 2
- 235000001674 Agaricus brunnescens Nutrition 0.000 description 2
- 206010003694 Atrophy Diseases 0.000 description 2
- 208000032544 Cicatrix Diseases 0.000 description 2
- 241000209140 Triticum Species 0.000 description 2
- 235000021307 Triticum Nutrition 0.000 description 2
- 239000012670 alkaline solution Substances 0.000 description 2
- 230000037444 atrophy Effects 0.000 description 2
- 239000002585 base Substances 0.000 description 2
- 230000004071 biological effect Effects 0.000 description 2
- 239000002131 composite material Substances 0.000 description 2
- 238000004042 decolorization Methods 0.000 description 2
- 238000004090 dissolution Methods 0.000 description 2
- 238000009826 distribution Methods 0.000 description 2
- IDGUHHHQCWSQLU-UHFFFAOYSA-N ethanol;hydrate Chemical compound O.CCO IDGUHHHQCWSQLU-UHFFFAOYSA-N 0.000 description 2
- 239000000835 fiber Substances 0.000 description 2
- 239000012530 fluid Substances 0.000 description 2
- 238000011081 inoculation Methods 0.000 description 2
- 239000002054 inoculum Substances 0.000 description 2
- 229920002521 macromolecule Polymers 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 230000000813 microbial effect Effects 0.000 description 2
- 239000011812 mixed powder Substances 0.000 description 2
- 239000000049 pigment Substances 0.000 description 2
- 238000001556 precipitation Methods 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 230000037387 scars Effects 0.000 description 2
- 239000013049 sediment Substances 0.000 description 2
- 238000003809 water extraction Methods 0.000 description 2
- FYGDTMLNYKFZSV-WFYNLLPOSA-N (2s,3r,4s,5s,6r)-2-[(2r,4r,5r,6s)-4,5-dihydroxy-2-(hydroxymethyl)-6-[(2r,3s,4r,5r,6s)-4,5,6-trihydroxy-2-(hydroxymethyl)oxan-3-yl]oxyoxan-3-yl]oxy-6-(hydroxymethyl)oxane-3,4,5-triol Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1[C@@H](CO)O[C@@H](O[C@@H]2[C@H](O[C@H](O)[C@H](O)[C@H]2O)CO)[C@H](O)[C@H]1O FYGDTMLNYKFZSV-WFYNLLPOSA-N 0.000 description 1
- 206010027145 Melanocytic naevus Diseases 0.000 description 1
- 208000007256 Nevus Diseases 0.000 description 1
- 244000131316 Panax pseudoginseng Species 0.000 description 1
- 235000005035 Panax pseudoginseng ssp. pseudoginseng Nutrition 0.000 description 1
- 235000003140 Panax quinquefolius Nutrition 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- 229930182558 Sterol Natural products 0.000 description 1
- 229930003427 Vitamin E Natural products 0.000 description 1
- 240000008042 Zea mays Species 0.000 description 1
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 1
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 239000002390 adhesive tape Substances 0.000 description 1
- 239000003513 alkali Substances 0.000 description 1
- 238000000498 ball milling Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 230000001276 controlling effect Effects 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 235000005822 corn Nutrition 0.000 description 1
- 230000009849 deactivation Effects 0.000 description 1
- 230000006837 decompression Effects 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 238000000502 dialysis Methods 0.000 description 1
- 235000013325 dietary fiber Nutrition 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 210000002615 epidermis Anatomy 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- WIGCFUFOHFEKBI-UHFFFAOYSA-N gamma-tocopherol Natural products CC(C)CCCC(C)CCCC(C)CCCC1CCC2C(C)C(O)C(C)C(C)C2O1 WIGCFUFOHFEKBI-UHFFFAOYSA-N 0.000 description 1
- 235000008434 ginseng Nutrition 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 230000000774 hypoallergenic effect Effects 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 235000004213 low-fat Nutrition 0.000 description 1
- 238000009629 microbiological culture Methods 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 235000012149 noodles Nutrition 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 229920001542 oligosaccharide Polymers 0.000 description 1
- 150000002482 oligosaccharides Chemical class 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 238000010298 pulverizing process Methods 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 239000002002 slurry Substances 0.000 description 1
- 238000002791 soaking Methods 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 150000003432 sterols Chemical class 0.000 description 1
- 235000003702 sterols Nutrition 0.000 description 1
- 238000010257 thawing Methods 0.000 description 1
- 239000011573 trace mineral Substances 0.000 description 1
- 235000013619 trace mineral Nutrition 0.000 description 1
- 150000004670 unsaturated fatty acids Chemical class 0.000 description 1
- 235000021122 unsaturated fatty acids Nutrition 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 229940046009 vitamin E Drugs 0.000 description 1
- 235000019165 vitamin E Nutrition 0.000 description 1
- 239000011709 vitamin E Substances 0.000 description 1
- 150000003722 vitamin derivatives Chemical class 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 238000001238 wet grinding Methods 0.000 description 1
- 229920001221 xylan Polymers 0.000 description 1
- 150000004823 xylans Chemical class 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/04—Polysaccharides, i.e. compounds containing more than five saccharide radicals attached to each other by glycosidic bonds
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/72—Cosmetics or similar toiletry preparations characterised by the composition containing organic macromolecular compounds
- A61K8/73—Polysaccharides
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/14—Preparation of compounds containing saccharide radicals produced by the action of a carbohydrase (EC 3.2.x), e.g. by alpha-amylase, e.g. by cellulase, hemicellulase
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
- C12P21/06—Preparation of peptides or proteins produced by the hydrolysis of a peptide bond, e.g. hydrolysate products
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
- A61K2800/84—Products or compounds obtained by lyophilisation, freeze-drying
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
- A61K2800/85—Products or compounds obtained by fermentation, e.g. yoghurt, beer, wine
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/645—Fungi ; Processes using fungi
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- General Health & Medical Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- General Chemical & Material Sciences (AREA)
- Biochemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Molecular Biology (AREA)
- Birds (AREA)
- Epidemiology (AREA)
- Dermatology (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The application discloses a product rich in beta-glucan and polypeptide, a preparation method and application thereof, wherein the preparation method comprises the following steps: s1, taking highland barley and/or oat rice as raw materials, crushing, sieving to obtain highland barley and/or oat rice powder, adding water, adjusting the pH value, adopting ultrasonic-assisted complex enzyme for enzymolysis, and inactivating enzyme to obtain an enzymolysis product; s2, inoculating an enzymolysis product into schizophyllum commune for fermentation, sterilizing and centrifuging to obtain a supernatant 1 and a precipitate; s3, performing high-pressure cell disruption extraction on the precipitate, centrifuging to remove residues, and obtaining supernatant 2; s4, combining the supernatant 1 and the supernatant 2, regulating the pH value, carrying out enzymolysis by adopting ultrasonic-assisted compound protease, separating and purifying, and drying to obtain a product rich in beta-glucan and polypeptide. The application combines the two ultrasonic-assisted enzymolysis and biological fermentation conversion technologies with the high-pressure cell disruption extraction technology, and has high efficiency, and the obtained product has high content of beta-glucan and polypeptide and good stability.
Description
Technical Field
The application relates to the technical field of cosmetics, in particular to a product rich in beta-glucan and polypeptide, a preparation method and application thereof.
Background
Highland barley, an important highland cereal crop, is rich in a large amount of dietary fibers, has nutritional ingredients higher than those of rice, wheat and corn, and has the characteristics of high protein, high fiber, high vitamin, low fat and low sugar, and is mainly produced in high-altitude areas such as Tibetan China and Qinghai. The content of beta-glucan in highland barley is the first place in wheat crops in the world, and the content of beta-glucan in the highland barley can reach 8.5 percent. Oat rice is one of miscellaneous cereals with high nutritional value in the world, is rich in nutrition, has higher effective components such as protein, unsaturated fatty acid, beta-glucan, active polypeptide and the like, and has the content of 8 amino acids and vitamin E required by human bodies higher than rice and white noodles.
Schizophyllum commune (academic name: schizophyllum commune Fr.) is also called white ginseng fungus, is fungus of Schizophyllum genus of Schizophyllum family, and is also rare mushroom fungus for both food and medicine. At present, most researches are carried out on extracellular polysaccharide generated by fermentation of schizophyllum commune, wherein the extracellular polysaccharide mainly comprises schizophyllum commune polysaccharide (SPG for short), also called schizophyllum commune, which is water-soluble beta-D-glucan with beta- (1-6) branches in a molecular structure; the cell wall and intracellular research on schizophyllum commune mycelium is relatively less, and the schizophyllum commune mainly contains relatively high protein, polysaccharides, amino acids and trace elements necessary for human bodies; the protein content and quality of schizophyllum commune are excellent protein sources, and the schizophyllum commune compound contains a plurality of active ingredients such as schizophyllum commune polysaccharide, schizophyllum commune, sterol and the like, so that the schizophyllum commune compound has remarkable medicinal value.
The patent researches on oat, highland barley and schizophyllum commune at present are as follows: chinese patent document CN 114292348A discloses an extraction method of oat polypeptide and beta-glucan and a nutritional product, wherein oat is processed by ball milling, so that the fiber structure of the oat is destroyed, the dissolution speed in the water extraction process of the beta-glucan is improved, and the yield of the beta-glucan is improved; meanwhile, the microwave-assisted water extraction method is adopted for extraction, and the rapid dissolution of beta-glucan and protein is realized by adjusting the power of microwaves; in addition, the beta-glucan is prepared by adopting a repeated freeze thawing method, so that the biological activity and purity of the beta-glucan are ensured, and the oat polypeptide is obtained by adopting an enzymolysis method, so that the yield of the oat polypeptide is higher, and the biological activity is better; chinese patent document CN114941019A discloses a method for reutilizing microbial fermentation residues, an extract of schizophyllum commune residues and application thereof, wherein the microbial fermentation residues are adjusted into bacterial liquid; performing wall breaking extraction on the bacterial liquid, and centrifuging to remove residues to obtain a first supernatant; concentrating the first supernatant to obtain a concentrated solution, performing alcohol precipitation on the concentrated solution, and performing centrifugal separation to obtain a precipitate and a second supernatant; adding complex enzyme into the second supernatant for enzymolysis; then separating and purifying to obtain a third supernatant; drying the precipitate to obtain a first extract SPE-SPG; concentrating the third supernatant to obtain an extract, and drying the extract to obtain a second extract SPE-SPP; chinese patent document CN 102585030A discloses a method for extracting beta-glucan by using grains, which comprises pulverizing grains or grains with seed coats removed, soaking in water to obtain a material with a water content of 40-85%, performing high-temperature high-pressure treatment, wet-milling to obtain a slurry, and performing enzymolysis with cellulase, alkali extraction, enzymolysis saccharification, and enzyme deactivation to obtain a feed liquid; finally, separating, purifying, concentrating and drying the feed liquid by adopting a membrane separation process to obtain beta-glucan powder; chinese patent document CN 107011460A discloses a method for extracting beta-glucan by taking highland barley bran as a raw material, and finally obtaining the highland barley bran beta-glucan by pretreating the highland barley bran raw material, removing starch and xylan, extracting alkaline solution, removing protein, dialyzing an extracting solution, concentrating and precipitating with ethanol, redissolving the precipitate and freeze-drying; the enzyme method and alkaline solution combined action are utilized to effectively improve the yield of beta-glucan, and the steps of dialysis and alcohol precipitation are adopted to effectively remove micromolecule and oligosaccharide impurities. Most of the prior art processes obtain low levels of beta-glucan or polypeptide by first enzymatic hydrolysis followed by purification; or the extracellular polysaccharide is obtained by a fermentation method except for protein, the protein in fermentation and the protein contained in mycelium are not hydrolyzed into polypeptide to be recovered, and the cell wall and the intracellular beta-glucan of the mycelium are recovered, so that the recovery rate is not high.
Disclosure of Invention
Aiming at the problems in the prior art, the application provides a product rich in beta-glucan and polypeptide, a preparation method and application thereof, and the conversion technology of twice ultrasonic auxiliary enzymolysis and biological fermentation with high efficiency and specificity is adopted, and simultaneously, the high-pressure cell disruption extraction technology is combined, so that the extraction rate of the beta-glucan and the polypeptide is improved, and the product of the beta-glucan and the polypeptide with high content, small molecular weight and good stability is obtained, thereby having good application prospect in the field of cosmetics.
The technical scheme of the application is as follows:
a product enriched in β -glucan and polypeptide, the method of making comprising the steps of:
s1, taking highland barley and/or oat rice as raw materials, crushing, sieving to obtain highland barley or oat rice powder, adding water, adjusting the pH value, adopting ultrasonic auxiliary complex enzyme for enzymolysis, and then inactivating enzyme to obtain an enzymolysis product;
s2, inoculating an enzymolysis product into a bacterial liquid of schizophyllum commune, fermenting, sterilizing under high pressure, and centrifuging to obtain a supernatant 1 and a precipitate;
s3, performing high-pressure cell disruption extraction on the precipitate, centrifuging to remove residues, and obtaining supernatant 2;
s4, combining the supernatant 1 and the supernatant 2, regulating the pH value, carrying out enzymolysis by adopting ultrasonic-assisted compound protease, separating and purifying, and freeze-drying to obtain a product rich in beta-glucan and polypeptide.
Further, the ultrasonic-assisted complex enzyme enzymolysis in the step S1 is carried out for 2-3 hours by adopting ultrasonic-assisted complex enzyme with the ultrasonic frequency of 300-500W and the temperature of 45-65 ℃.
Furthermore, the compound enzyme in the step S1 is cellulase, amylase and papain, and the addition amount of the cellulase, amylase and papain is (1% -2%), 0.5% -1% and 0.5% -1% of the total weight of highland barley and/or oat rice powder.
Further, the step S2 of inoculating the schizophyllum commune seed solution refers to inoculating the schizophyllum commune seed solution with 10-20% of the inoculation amount of the schizophyllum commune seed solution into the enzymolysis product after culturing the schizophyllum commune in the seed culture solution for 3 days.
Further, the fermentation in the step S2 is carried out for 5-7 days at the temperature of 25-28 ℃ and the rotating speed of 100-200 rpm/min.
Further, the high-pressure cell disruption extraction in the step S3 refers to the high-pressure cell disruption extraction for 10-30min under the conditions that the pressure is 1000-2000bar and the temperature is 20-40 ℃.
Further, the compound protease in the step S4 comprises papain, subtilisin and trypsin, and the addition amounts of the papain, the subtilisin and the trypsin are respectively (0.5% -1%), the (0.2% -0.5%) and the (0.2% -5%) of the combined total weight of the supernatant 1 and the supernatant 2.
Further, the separation and purification in the step S4 means that 0.5-1% of active carbon of the total combined weight of the supernatant 1 and the supernatant 2 is decolorized, and the mixture is filtered by a 10 mu m polypropylene micro-filtration membrane and then separated by a Sephadex G-25 gel chromatographic column.
Further, the average molecular weight of the β -glucan and polypeptide rich product obtained in step S4 is less than 5kDa.
Further, the product rich in beta-glucan and polypeptide obtained in the step S4 is dissolved and precipitated by adopting an ethanol water solution with the concentration of 60-80%, and is kept stand for 12 hours, and is centrifuged for 10-20 min at 3000-5000 rpm, so that the purity of the product which is precipitated to be rich in beta-glucan is more than 75%, the supernatant is subjected to low-temperature decompression to recover ethanol, and the product which is rich in polypeptide is obtained after drying is more than 60%.
The application also provides an application of the product rich in beta-glucan and polypeptide in preparing cosmetics.
The principle of the application is as follows: the application adopts first complex enzyme enzymolysis, and obtains full release of nutrient components of highland barley and/or oat rice under the assistance of ultrasonic waves, then is inoculated into schizophyllum for fermentation, macromolecular substances are further converted into micromolecular substances, simultaneously substances such as schizophyllum extracellular polysaccharide, schizophyllum intracellular polysaccharide, amino acid and the like are produced by the fermentation of the schizophyllum, the content of schizophyllum beta-glucan and polypeptide is increased, mycelium, highland barley rice residue and/or oat rice residue and the like are extracted by using a high-pressure cell disruption extraction technology, the components of substances such as schizophyllum wall and substances for promoting intracellular polysaccharide and protein are released, the content of schizophyllum beta-glucan and polypeptide is further increased, then the second complex protease enzymolysis is adopted, the proteins after fermentation and mycelium are directionally hydrolyzed into polypeptides after high-pressure cell disruption, the content of polypeptides is further increased, finally, the decolorization and filtration are carried out, and the beta-glucan and the polypeptides are separated and purified by a Sephadex G-25 gel chromatographic column, so that the products with high content, small molecular weight and good stability are obtained.
Compared with the prior art, the application has the beneficial effects that:
1. according to the application, the nutrition characteristics of highland barley and/or oat rice are fully utilized, the composite enzyme enzymolysis is firstly adopted to fully release the nutrition components of highland barley and/or oat rice under the assistance of ultrasonic waves, then the schizophyllum is inoculated to ferment to further convert macromolecular substances into micromolecular substances, simultaneously the schizophyllum is used to ferment to generate substances such as schizophyllum extracellular polysaccharide, schizophyllum intracellular polysaccharide and amino acid, the content of schizophyllum beta-glucan and polypeptide is increased, then the substances such as mycelium, highland barley residue and/or oat rice residue are extracted by using a high-pressure cell disruption extraction technology to release the components of the substances such as schizophyllum wall, intracellular polysaccharide and protein, the content of schizophyllum beta-glucan and polypeptide is increased, the enzymolysis is carried out by using composite protease, the proteins after fermentation and mycelium are directionally hydrolyzed into the polypeptides after high-pressure cell disruption, the polypeptide content is further increased, finally the decolorization is carried out, the products of beta-glucan and the polypeptides with high content, small molecular weight and good stability are obtained by separating by using a Sephadex G-25 gel chromatographic column, and the preparation method is high in extraction rate and/or the effective nutrition components of highland oat rice can be fully utilized.
2. The beta-glucan and polypeptide-rich product prepared by the application not only contains the beta-glucan and polypeptide products of highland barley and/or oat rice, but also contains the schizophyllum beta-glucan and polypeptide products of the schizophyllum commune which are fermented to produce rare mushroom fungi, and meanwhile, the beta-glucan and polypeptide-rich product has small molecular weight and is a small molecular beta-glucan and polypeptide product, and the preparation utilization rate is high.
Drawings
FIG. 1 is a flow chart of the production process of example 1 of the present application;
FIG. 2 is a diagram of a strain liquid after 3 days of liquid culture of schizophyllum commune of example 1 of the present application, a diagram of a fermented product after 5 days of fermentation culture, a diagram of a liquid after high-pressure cell wall breaking of a precipitate (including mycelium, highland barley rice residue, oat rice residue, etc.), a diagram of a liquid supernatant 1, a diagram of a liquid supernatant 2, and a diagram of a liquid of a product enriched in beta-glucan and polypeptide obtained after separation and purification by a gel chromatography column.
Detailed Description
The present application will be further described in detail with reference to the following embodiments, in order to make the objects, technical solutions and advantages of the present application more apparent. It should be understood that the description is only illustrative and is not intended to limit the scope of the application. In addition, in the following description, descriptions of well-known structures and techniques are omitted so as not to unnecessarily obscure the present application.
For a better illustration of the application, typical but non-limiting embodiments thereof are described below in order to facilitate the understanding of the technical solutions of the application. The enzyme preparation used in the application is purchased from Nanning east Henghua biological technology limited public responsibility company, schizophyllum commune Schizophyllum commune is purchased from China industry microbiological culture Collection center, and the code: the sources of the other raw materials used for CICC2592 are not particularly limited, and they may be commercially available or may be prepared according to conventional methods well known to those skilled in the art.
EXAMPLE 1 beta-glucan and polypeptide-enriched product of the application and its preparation
The preparation of the product enriched in β -glucan and polypeptide of this example comprises the steps of:
s1, mixing 20g of highland barley and 20g of oat as raw materials, crushing, sieving with a 80-mesh sieve to obtain highland barley and oat mixed powder, wherein the powder is as follows: adding water in a mass ratio of 1:15, regulating the pH value to 6.0, performing enzymolysis by adopting ultrasonic-assisted compound enzyme with ultrasonic frequency of 500W and temperature of 45 ℃, wherein the compound enzyme consists of cellulase, amylase and papain, the addition amount of the three is 2%, 0.5% and 0.5% of the total weight of the mixed powder, namely the mass ratio of the cellulase, amylase and papain is 4:1:1, performing enzymolysis for 3 hours, and inactivating the enzyme at 90 ℃ to obtain an enzymolysis product;
s2, culturing schizophyllum commune in a seed culture solution for 3 days, inoculating schizophyllum commune strain solution with an inoculum size of 15% of the weight of an enzymolysis product into the enzymolysis product, controlling the temperature to be 28 ℃, fermenting at 160rpm/min for 5 days, sterilizing at 121 ℃ for 20min under high pressure after fermentation, and centrifuging at 5000rpm for 10min to obtain supernatant 1 and precipitate;
s3, adding sterile water with the mass 5 times of that of the precipitate into the precipitate, crushing and extracting the precipitate for 20min at the pressure of 1500bar and the temperature of 30 ℃, and centrifuging at 5000rpm for 10min to remove residues, thereby obtaining supernatant 2;
s4, combining the supernatant 1 and the supernatant 2, regulating the pH value to be 6.0, adopting ultrasonic wave with the frequency of 500W and the temperature of 45 ℃ to assist compound protease, wherein the compound protease comprises papain, subtilisin and trypsin, the adding amount of the papain, the subtilisin and the trypsin is respectively 1 percent, 0.2 percent and 0.2 percent of the total combined weight of the supernatant 1 and the supernatant 2, namely the mass ratio of the papain, the subtilisin and the trypsin is 5:1:1, carrying out enzymolysis for 3 hours, adding activated carbon with the total combined weight of the supernatant 1 and the supernatant 2 for decoloring, filtering by using a 10 mu m polypropylene microfiltration membrane, separating and purifying by using a Sephadex G-25 gel chromatographic column, and carrying out freeze drying to obtain the product rich in beta-glucan and polypeptide. Dissolving and precipitating the product rich in beta-glucan and polypeptide by adopting 75% ethanol water solution, standing for 12 hours, centrifuging at 5000rpm for 10min to obtain the product rich in beta-glucan, recovering ethanol from supernatant at low temperature and under reduced pressure, and drying to obtain the product rich in polypeptide. Referring to fig. 1, a production process flow chart of the present embodiment is shown; referring to fig. 2, a strain liquid diagram of schizophyllum commune liquid culture for 3 days, a ferment liquid diagram of schizophyllum commune liquid culture for 5 days, a liquid diagram of sediment (containing mycelium, highland barley rice residue, oat rice residue and other substances) subjected to high-pressure cell wall breaking, a liquid diagram of supernatant 1, a liquid diagram of supernatant 2, and a liquid diagram of a product rich in beta-glucan and polypeptide obtained after gel chromatographic column separation and purification are sequentially shown from left to right.
EXAMPLE 2 beta-glucan and polypeptide-enriched products of the application and their preparation
The preparation of the product enriched in β -glucan and polypeptide of this example comprises the steps of:
s1, taking 40g of highland barley as a raw material, crushing, sieving with a 150-mesh sieve to obtain highland barley powder, and mixing the powder with the following components: adding water in a mass ratio of 1:10, adjusting the pH value to 5.0, and then adopting ultrasonic wave with the frequency of 300W and the temperature of 50 ℃ to assist enzymolysis by using ultrasonic-assisted compound enzyme, wherein the compound enzyme consists of cellulase, amylase and papain, and the adding amount of the cellulase, the amylase and the papain is 1 percent, 1 percent and 1 percent of the weight of highland barley flour powder, namely the mass ratio of the cellulase, the amylase and the papain is 1:1:1, performing enzymolysis for 2 hours, and inactivating enzyme at 80 ℃ to obtain an enzymolysis product;
s2, after the schizophyllum commune is cultured in a seed culture solution for 3 days, the schizophyllum commune strain solution with the inoculation amount of 10 percent of the weight of an enzymolysis product is inoculated in the enzymolysis product, the temperature is controlled to be 28 ℃, the rotating speed is 160rpm/min, the fermentation is carried out for 7 days, after the fermentation is finished, the high-pressure sterilization is carried out for 20min at 121 ℃, and the centrifugation is carried out for 20min at 3000rpm, so as to obtain supernatant 1 and sediment;
s3, adding sterile water with the mass 10 times of that of the precipitate into the precipitate, crushing and extracting the precipitate for 10min at the pressure of 2000bar and the temperature of 20 ℃, and centrifuging at 3000rpm for 20min to remove residues, thereby obtaining supernatant 2;
s4, combining the supernatant 1 and the supernatant 2, regulating the pH value to 7.0, performing enzymolysis by adopting ultrasonic-assisted compound protease with the ultrasonic frequency of 300W and the temperature of 50 ℃, wherein the compound protease comprises papain, subtilisin and trypsin, the adding amount of the papain, the subtilisin and the trypsin is respectively 0.7 percent, 0.2 percent and 0.5 percent of the total combined weight of the supernatant 1 and the supernatant 2, namely the mass ratio of the papain to the subtilisin to the trypsin is 7:2:5, performing enzymolysis for 2 hours, adding activated carbon with the total combined weight of the supernatant 1 and the supernatant 2 to decolor, filtering by using a 10 mu m polypropylene microfiltration membrane, separating and purifying by using a Sephadex G-25 gel chromatographic column, and performing freeze drying to obtain the product rich in beta-glucan and polypeptide. Dissolving and precipitating the product rich in beta-glucan and polypeptide by adopting 70% ethanol aqueous solution, standing for 12 hours, centrifuging at 4000rpm for 15min to obtain the product which is precipitated to be rich in beta-glucan, recovering ethanol from supernatant fluid at low temperature and under reduced pressure, and drying to obtain the product rich in polypeptide.
EXAMPLE 3 beta-glucan and polypeptide-enriched products of the application and their preparation
The preparation of the product enriched in β -glucan and polypeptide of this example comprises the steps of:
s1, taking 40g of oat as a raw material, crushing, sieving with a 200-mesh sieve to obtain oat powder, and mixing the powder with the following components: adding water in a mass ratio of 1:20, adjusting the pH value to 7.0, and then adopting ultrasonic wave with the frequency of 300W and the temperature of 55 ℃ to assist enzymolysis by using compound enzyme, wherein the compound enzyme consists of cellulase, amylase and papain, and the adding amount of the cellulase, the amylase and the papain is 1.5%, 0.75% and 0.75% of the total weight of the oat flour, namely the mass ratio of the cellulase, the amylase and the papain is 2:1:1, performing enzymolysis for 2 hours, and inactivating enzyme at 80 ℃ to obtain an enzymolysis product;
s2, culturing schizophyllum commune in a seed culture solution for 3 days, inoculating schizophyllum commune strain solution with an inoculum size of 20% of the mass of an enzymolysis product into the enzymolysis product, fermenting at a temperature of 25 ℃ and a rotating speed of 160rpm/min for 7 days, sterilizing at 121 ℃ for 20min under high pressure after fermentation, and centrifuging at 4000rpm for 15min to obtain a supernatant 1 and a precipitate;
s3, adding sterile water with the mass 10 times of that of the precipitate into the precipitate, crushing and extracting the precipitate for 30min at the high-pressure cell with the pressure of 1000pa and the temperature of 40 ℃, centrifuging at 4000rpm for 15min, centrifuging and deslagging to obtain supernatant 2;
s4, combining the supernatant 1 and the supernatant 2, regulating the pH value to be 5.0, performing enzymolysis by adopting ultrasonic-assisted compound protease with the ultrasonic frequency of 300W and the temperature of 50 ℃, wherein the compound protease comprises papain, subtilisin and trypsin, the adding amount of the papain, the subtilisin and the trypsin is respectively 0.7 percent, 0.5 percent and 0.2 percent of the total combined weight of the supernatant 1 and the supernatant 2, namely, the mass ratio of the papain, the subtilisin and the trypsin is 7:5:2, performing enzymolysis for 2 hours, decoloring by using activated carbon with the total combined weight of the supernatant 1 and the supernatant 2 of 1 percent, filtering by using a 10 mu m polypropylene microfiltration membrane, separating and purifying by using a Sephadex G-25 gel chromatographic column, and performing freeze drying to obtain the product rich in beta-glucan and polypeptide. Dissolving and precipitating the product rich in beta-glucan and polypeptide by adopting an aqueous solution of ethanol with the concentration of 80%, standing for 12 hours, centrifuging at 3000rpm for 20min to obtain the product which is precipitated to be rich in beta-glucan, recovering ethanol from supernatant fluid at low temperature and under reduced pressure, and drying to obtain the product rich in polypeptide.
Comparative example 1
The comparative example is different from example 1 only in that the ultrasonic-assisted complex enzyme hydrolysis is not performed in step S1, and the other is exactly the same as example 1.
Comparative example 2
The comparative example is different from example 1 only in that the mass ratio of cellulase, amylase and papain in the ultrasonic-assisted enzyme complex in step S1 is 3:2:1, and the other is exactly the same as example 1.
Comparative example 3
This comparative example was different from example 1 only in that fermentation was performed without adding schizophyllum commune strain in step S2, and the other steps were exactly the same as in example 1.
Comparative example 4
This comparative example is different from example 1 only in that the precipitate is not subjected to high-pressure cell wall-breaking extraction in step S3, and the other is exactly the same as example 1.
Comparative example 5
This comparative example is different from example 1 only in that the ultrasonic-assisted complex protease is not performed in step S4, and the other is exactly the same as example 1.
Comparative example 6
The comparative example is different from example 1 only in that the mass ratio of papain to subtilisin to trypsin added in the ultrasonic-assisted complex protease enzymatic hydrolysis in step S4 is 3:2:2.
Comparative example 7
This comparative example is different from example 1 only in that fermentation is performed without adding schizophyllum commune strain in step S2 and high-pressure cell wall breaking extraction is not performed on the precipitate in step S3, and the other is exactly the same as example 1.
Comparative example 8
This comparative example is different from example 1 only in that the step S3 is not performed for high pressure cell wall breaking extraction in the precipitate and the step S4 is not performed for the ultrasonic-assisted complex protease, and the other is exactly the same as example 1.
Test example 1 determination of beta-glucan content, beta-glucan content and molecular weight
1. Test sample: the products enriched in beta-glucan and polypeptide prepared in examples 1-3, comparative examples 1-8;
2. the test method comprises the following steps: measurement of beta-glucan content reference NY/T2006-2011 "measurement of beta-glucan content in cereal and products thereof; molecular weight distribution measurement is described in GB/T22492-2008 appendix A, method for measuring relative molecular weight distribution of peptide; the method for measuring the content of the polypeptide refers to a method for measuring the content of the peptide in GB/T22492-2008 annex B;
3. test results:
table 1, table of beta-glucan content, polypeptide content and molecular weight measurement results
As can be seen from the results in Table 1, the beta-glucan and polypeptide-enriched product samples prepared in examples 1-3 of the present application have the same content of beta-glucan and polypeptide and the average molecular weight significantly better than those of the samples prepared in comparative examples 1-6, which means that the beta-glucan and polypeptide-enriched product prepared in the present application not only contains the beta-glucan and polypeptide from highland barley and/or oat, but also increases the beta-glucan and polypeptide-enriched product from schizophyllum commune fermentation to produce the beta-glucan and polypeptide from the rare fungus species, and the average molecular weight gradually decreases along with the enzymolysis and fermentation, so that the beta-glucan and polypeptide-enriched product with high content and small average molecular weight can be obtained; the content and average molecular weight of the beta-glucan and the polypeptide of example 1 are both optimal, and are the best examples of the application. The comparative examples 1-8 changed the mass ratio of the ultrasonic-assisted complex enzyme enzymolysis and complex enzyme in the step S1, the schizophyllum commune fermentation step in the step S2, the high-pressure cell disruption and extraction technology in the step S3, and the complex protease enzymolysis and complex protease mass ratio in the step S4, all affecting the content and average molecular weight of the beta-glucan and the polypeptide of the finally obtained beta-glucan and polypeptide product.
Test example 2, moisture efficacy test
1. Test article: the products rich in beta-glucan and polypeptide prepared in examples 1-3 and the prepared samples of comparative examples 1-8 were prepared as 1.0% (w/w) aqueous solutions, and 10% (w/w) aqueous glycerol solution was used as a positive control;
2. the test method comprises the following steps: 24 healthy men and women were selected, 12 for each, and randomly divided into 12 groups of 1 for each of men and women. Before the test, the hands and arms of the test subjects are required to enter a climate control chamber (22+/-1 ℃ C., relative humidity 50%) for more than 20 minutes after washing, and the test subjects are kept in a relaxed state, and the forearms are free from scars, pigments, atrophy, moles of scars or other flaws and the like, which influence the test result judgment. Randomly marking 3 skins with the area of 3cm multiplied by 3cm on the inner sides of the left forearm and the right forearm of a subject as test areas, wherein the smearing amount of the inner sides of the left hand and the right hand of each subject is (2.0+/-0.1) mg/cm 2 Examples 1 to 3, comparative examples 1 to 8, and positive control, skin water was performed at time intervals of 1h, 2h, 4h, 6hThe content testing is carried out by adopting a skin moisture content Corneometer test probe of a multifunctional skin tester of the Germany CK company to test the skin moisture content, calculating the skin moisture content variation and calculating the average value;
3. test results:
TABLE 2 average skin moisture content variation
From the test results in Table 2, it is understood that the moisturizing effect of the products rich in beta-glucan and polypeptide prepared in examples 1 to 3 of example 1 is significantly higher than that of the prepared samples of comparative examples 1 to 8, when aqueous solutions (1.0%) at the same concentration are prepared; the product aqueous solution of the beta-glucan and the polypeptide in the examples 1-3 has equivalent moisturizing effect to that of the positive control group (10% glycerol aqueous solution) in 1-6 hours, which indicates that the product rich in the beta-glucan and the polypeptide prepared by the application has good moisturizing effect on skin, and if the mass ratio of the ultrasonic-assisted complex enzyme enzymolysis and the complex enzyme in the step S1, the schizophyllum commune fermentation step in the step S2, the high-pressure cell disruption and extraction technology in the step S3 and the mass ratio of the complex protease enzymolysis and the complex protease in the step S4 are changed, the content and the average molecular weight of the beta-glucan and the polypeptide of the finally obtained product of the beta-glucan and the polypeptide are influenced, so that the moisturizing effect on skin is influenced.
Test example 3 efficacy test for repairing skin Barrier
1. Test sample: the products rich in beta-glucan and polypeptide prepared in examples 1-3 and the prepared samples of comparative examples 1-8 were prepared as 1.0% (w/w) aqueous solutions, and 10% (w/w) aqueous glycerol solution was used as a positive control;
2. the test method comprises the following steps:
(1) preparation before testing: the volunteers are selected by the subjects, and are randomly divided into 12 groups of 24 healthy men and women, wherein each group of men and women comprises 1 person, and the volunteers have no scar, pigment, atrophy, bright red nevus or other flaws on the forearms, which affect the test result judgment, and the ages of the volunteers are between 20 and 40 years;
(2) other laboratory instruments and materials: hypoallergenic external tape (3M company, usa), percutaneous moisture loss meter VapoMeter (model SWL 5648);
(3) the experimental method comprises the following steps: skin with an area of 3cm×3cm was randomly marked as a test area on the inner sides of the left and right forearms of each subject, and was marked as a blank group (without using any sample) and an experimental group (using examples 1 to 3 and comparative examples 1 to 8, positive control), skin percutaneous moisture loss value (TEWL value) was measured as a base value before modeling, denoted as T0, tear-pull treatment was performed with a transparent adhesive tape in the test areas of the experimental group and the blank group, respectively, skin percutaneous moisture loss value (TEWL value) was measured again, and as a base value after modeling, denoted as T1. Using a test sample in a test area of the experimental group, using no sample in a blank control group, measuring the skin percutaneous moisture loss value (TEWL value) again after sitting still for 1h in a constant temperature and humidity environment, taking the measured skin percutaneous moisture loss value as a modeled repair value, marking as T2, and calculating skin repair rate (%) = (T1-T2)/(T1-T0) 100% after using the sample for 1 h;
3. test results:
TABLE 3 results of efficacy test for skin Barrier repair
As can be seen from Table 3, the test samples of the products rich in β -glucan and polypeptide prepared in examples 1-3 of the present application have significantly higher repair rates after being applied to the skin for 1 hour than the samples prepared in comparative examples 1-8 and the positive control group, which indicate that the products rich in β -glucan and polypeptide prepared in the present application can repair damaged skin barriers, and have an effect of reducing epidermis water loss, and if the mass ratio of the ultrasonic-assisted complex enzyme enzymatic hydrolysis and complex enzyme in step S1, the schizophyllum fermentation step in step S2, the high-pressure cell disruption extraction technique in step S3, the complex protease enzymatic hydrolysis and complex protease in step S4 are changed, the contents and the average molecular weights of the β -glucan and the polypeptide of the finally obtained products of β -glucan and polypeptide are all affected, thereby affecting the efficacy of repairing damaged skin barriers.
The above examples illustrate only a few embodiments of the application, which are described in detail and are not to be construed as limiting the scope of the application. It should be noted that it will be apparent to those skilled in the art that several variations and modifications can be made without departing from the spirit of the application, which are all within the scope of the application. Accordingly, the scope of protection of the present application is to be determined by the appended claims.
Claims (10)
1. A beta-glucan and polypeptide-rich product, wherein the preparation method of the beta-glucan and polypeptide-rich product comprises the steps of:
s1, taking highland barley and/or oat rice as raw materials, crushing, sieving to obtain highland barley and/or oat rice powder, adding water, adjusting the pH value, adopting ultrasonic-assisted complex enzyme for enzymolysis, and then inactivating enzyme to obtain an enzymolysis product;
s2, inoculating an enzymolysis product into a bacterial liquid of schizophyllum commune, fermenting, sterilizing under high pressure, and centrifuging to obtain a supernatant 1 and a precipitate;
s3, performing high-pressure cell disruption extraction on the precipitate, centrifuging to remove residues, and obtaining supernatant 2;
s4, combining the supernatant 1 and the supernatant 2, regulating the pH value, carrying out directional enzymolysis by adopting ultrasonic-assisted compound protease, decoloring, separating and purifying, and freeze-drying to obtain a product rich in beta-glucan and polypeptide.
2. The product rich in beta-glucan and polypeptide according to claim 1 wherein the ultrasound-assisted complex enzyme in step S1 is enzymatically hydrolyzed using an ultrasound-assisted complex enzyme having an ultrasound frequency of 300-500W and a temperature of 45-65 ℃ for 2-3 hours.
3. The product rich in beta-glucan and polypeptide according to claim 1 wherein the complex enzymes in step S1 are cellulase, amylase and papain and the amounts added are (1% -2%), (0.5% -1%) and (0.5% -1%) of the total weight of highland barley and/or oat flour.
4. The product rich in beta-glucan and polypeptide according to claim 1, wherein the step S2 of inoculating the schizophyllum commune seed solution is to inoculate the schizophyllum commune seed solution with 10-20% of the schizophyllum commune seed solution of the enzymatic hydrolysate after 3 days of culture of the schizophyllum commune seed solution.
5. The product enriched in β -glucan and polypeptide as claimed in claim 1 wherein the fermentation in step S2 is performed at a temperature of 25-28 ℃ and a rotational speed of 100-200 rpm/min for 5-7 days.
6. The product rich in β -glucan and polypeptide according to claim 1 wherein the high pressure cell disruption extraction in step S3 is a high pressure cell disruption extraction at a pressure of 1000-2000bar and a temperature of 20-40 ℃ for 10-30min.
7. The beta-glucan and polypeptide-rich product of claim 1 wherein the complex protease of step S4 comprises papain, subtilisin and trypsin, and the three are added in amounts of (0.5% -1%), (0.2% -0.5%) and (0.2% -5%) of the combined total weight of supernatant 1 and supernatant 2, respectively.
8. The product rich in beta-glucan and polypeptide according to claim 1 wherein the separation and purification in step S4 is performed by decolorizing with activated carbon in an amount of 0.5-1% of the combined total weight of supernatant 1 and supernatant 2, filtering with 10 μm polypropylene microfiltration membrane, and separating with Sephadex G-25 gel chromatography column.
9. The product rich in beta-glucan and polypeptide according to claim 1 wherein the product rich in beta-glucan and polypeptide obtained in step S4 is obtained by dissolving and precipitating with ethanol in 60-80% ethanol aqueous solution, standing for 12 hours, centrifuging at 3000-5000 rpm for 10-20 min to obtain a product with purity of more than 75% precipitated to be rich in beta-glucan, recovering ethanol from the supernatant at low temperature under reduced pressure, and drying to obtain a product with purity of more than 60% enriched in polypeptide.
10. Use of a product enriched in β -glucan and polypeptide as claimed in any one of claims 1-9 in the preparation of a cosmetic.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310702412.9A CN116694702A (en) | 2023-06-13 | 2023-06-13 | Product rich in beta-glucan and polypeptide as well as preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310702412.9A CN116694702A (en) | 2023-06-13 | 2023-06-13 | Product rich in beta-glucan and polypeptide as well as preparation method and application thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN116694702A true CN116694702A (en) | 2023-09-05 |
Family
ID=87827248
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202310702412.9A Pending CN116694702A (en) | 2023-06-13 | 2023-06-13 | Product rich in beta-glucan and polypeptide as well as preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN116694702A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117959208A (en) * | 2024-01-29 | 2024-05-03 | 广州星汇生物科技有限公司 | Multiple beta-glucan oat fermentation liquor with moisturizing and anti-wrinkle effects and preparation method and application thereof |
-
2023
- 2023-06-13 CN CN202310702412.9A patent/CN116694702A/en active Pending
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117959208A (en) * | 2024-01-29 | 2024-05-03 | 广州星汇生物科技有限公司 | Multiple beta-glucan oat fermentation liquor with moisturizing and anti-wrinkle effects and preparation method and application thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN108157579B (en) | Preparation method of cardamine violifolia selenium polypeptide with high organic selenium content | |
| CN106244658B (en) | Preparation method of sweet potato protein polypeptide | |
| CN108324633A (en) | A kind of preparation method and products thereof of fermentative brown rice stoste used for cosmetic | |
| CN116694702A (en) | Product rich in beta-glucan and polypeptide as well as preparation method and application thereof | |
| CN113755544B (en) | Schizophyllum commune fermentation product, and preparation method and application thereof | |
| CN107541533A (en) | A kind of preparation method of medicine food hypha polysaccharide polypeptide immunopotentiator | |
| WO2020181833A1 (en) | Method for extracting selenoprotein from selenium-rich edible mushroom | |
| CN108034688A (en) | The preparation process of rice bran polysaccharide | |
| CN111334540A (en) | Method for extracting dendrobium officinale polysaccharide by utilizing biological fermentation | |
| CN109170922B (en) | Preparation method of wheat bran soluble dietary fiber | |
| CN111387489B (en) | Lactic acid bacteria bird's nest product and preparation method thereof | |
| CN103229666B (en) | Cordyceps militaris peanut and preparing method thereof | |
| CN108588142B (en) | Method for improving polysaccharide content of ganoderma lucidum mycelia by utilizing fungal polysaccharide and ganoderma lucidum product obtained by method | |
| CN112961883B (en) | Heat-resistant white kidney bean amylase inhibitor and preparation method thereof | |
| CN111418827B (en) | Lactobacillus plantarum with functions of improving enzymolysis and fermentation flavor of bird's nest and increasing small molecular peptides and sialic acid | |
| CN114177121A (en) | Preparation method and application of pollen pini probiotic fermented cosmetic raw material | |
| CN106819778A (en) | A kind of preparation method of water-soluble dietary fiber of corn peels | |
| CN112522114A (en) | Cordyceps militaris mushroom residue extracting solution, lucid ganoderma fermentation product, preparation method and application thereof | |
| CN114150033B (en) | Preparation method of rice bran antioxidant peptide compound treated by peak alpha amylase | |
| CN112899213B (en) | Nutrition promoter and application thereof in hericium erinaceus fermentation culture | |
| CN110447720B (en) | Deep processing method of red dates | |
| CN117286206B (en) | Enzyme-ferment coupling preparation method of wolfberry glycosylated polypeptide | |
| CN117618316B (en) | Fermentation composition, mask and preparation method and application thereof | |
| CN113080453A (en) | Preparation method of special dietary supplement | |
| CN106889631A (en) | A kind of preparation method of oat kind severe edema due to hypofunction of the spleen soluble dietary fiber |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |