CN112708648A - Preparation method and application of pharmaceutical-grade sparassis crispa polysaccharide for enhancing immunity - Google Patents
Preparation method and application of pharmaceutical-grade sparassis crispa polysaccharide for enhancing immunity Download PDFInfo
- Publication number
- CN112708648A CN112708648A CN202011561116.4A CN202011561116A CN112708648A CN 112708648 A CN112708648 A CN 112708648A CN 202011561116 A CN202011561116 A CN 202011561116A CN 112708648 A CN112708648 A CN 112708648A
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- Prior art keywords
- parts
- sparassis crispa
- polysaccharide
- powder
- ionic liquid
- Prior art date
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- Pending
Links
- 241000272503 Sparassis radicata Species 0.000 title claims abstract description 121
- 150000004676 glycans Chemical class 0.000 title claims abstract description 95
- 229920001282 polysaccharide Polymers 0.000 title claims abstract description 95
- 239000005017 polysaccharide Substances 0.000 title claims abstract description 95
- 230000036039 immunity Effects 0.000 title claims abstract description 29
- 230000002708 enhancing effect Effects 0.000 title claims abstract description 21
- 238000002360 preparation method Methods 0.000 title claims abstract description 14
- 102000004190 Enzymes Human genes 0.000 claims abstract description 56
- 108090000790 Enzymes Proteins 0.000 claims abstract description 56
- 239000002608 ionic liquid Substances 0.000 claims abstract description 44
- 238000003756 stirring Methods 0.000 claims abstract description 25
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 18
- 239000002994 raw material Substances 0.000 claims abstract description 18
- 239000000706 filtrate Substances 0.000 claims abstract description 11
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- 230000007062 hydrolysis Effects 0.000 claims abstract description 7
- 238000006460 hydrolysis reaction Methods 0.000 claims abstract description 7
- 238000001035 drying Methods 0.000 claims abstract description 6
- 238000001914 filtration Methods 0.000 claims abstract description 6
- 108010009736 Protein Hydrolysates Proteins 0.000 claims abstract description 3
- 229940088598 enzyme Drugs 0.000 claims description 53
- 239000000843 powder Substances 0.000 claims description 50
- 239000000243 solution Substances 0.000 claims description 33
- 239000011259 mixed solution Substances 0.000 claims description 24
- 239000007788 liquid Substances 0.000 claims description 18
- 238000000034 method Methods 0.000 claims description 15
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- 108010084185 Cellulases Proteins 0.000 claims description 12
- 235000014486 Hydrangea macrophylla Nutrition 0.000 claims description 11
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- 102000016943 Muramidase Human genes 0.000 claims description 11
- 108010062010 N-Acetylmuramoyl-L-alanine Amidase Proteins 0.000 claims description 11
- 239000007864 aqueous solution Substances 0.000 claims description 11
- 235000013305 food Nutrition 0.000 claims description 11
- 229960000274 lysozyme Drugs 0.000 claims description 11
- 239000004325 lysozyme Substances 0.000 claims description 11
- 235000010335 lysozyme Nutrition 0.000 claims description 11
- 239000007787 solid Substances 0.000 claims description 11
- 239000000126 substance Substances 0.000 claims description 10
- WVXRAFOPTSTNLL-NKWVEPMBSA-N 2',3'-dideoxyadenosine Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@H]1CC[C@@H](CO)O1 WVXRAFOPTSTNLL-NKWVEPMBSA-N 0.000 claims description 8
- 229920001202 Inulin Polymers 0.000 claims description 8
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- 102000004882 Lipase Human genes 0.000 claims description 8
- 108090001060 Lipase Proteins 0.000 claims description 8
- 240000003394 Malpighia glabra Species 0.000 claims description 8
- 235000014837 Malpighia glabra Nutrition 0.000 claims description 8
- 241000208829 Sambucus Species 0.000 claims description 8
- 235000018735 Sambucus canadensis Nutrition 0.000 claims description 8
- 108010073771 Soybean Proteins Proteins 0.000 claims description 8
- 108010046377 Whey Proteins Proteins 0.000 claims description 8
- 102000007544 Whey Proteins Human genes 0.000 claims description 8
- 235000007123 blue elder Nutrition 0.000 claims description 8
- 239000005018 casein Substances 0.000 claims description 8
- BECPQYXYKAMYBN-UHFFFAOYSA-N casein, tech. Chemical compound NCCCCC(C(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(CC(C)C)N=C(O)C(CCC(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(C(C)O)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(COP(O)(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(N)CC1=CC=CC=C1 BECPQYXYKAMYBN-UHFFFAOYSA-N 0.000 claims description 8
- 235000021240 caseins Nutrition 0.000 claims description 8
- 235000007124 elderberry Nutrition 0.000 claims description 8
- 235000008995 european elder Nutrition 0.000 claims description 8
- FTSSQIKWUOOEGC-RULYVFMPSA-N fructooligosaccharide Chemical compound OC[C@H]1O[C@@](CO)(OC[C@@]2(OC[C@@]3(OC[C@@]4(OC[C@@]5(OC[C@@]6(OC[C@@]7(OC[C@@]8(OC[C@@]9(OC[C@@]%10(OC[C@@]%11(O[C@H]%12O[C@H](CO)[C@@H](O)[C@H](O)[C@H]%12O)O[C@H](CO)[C@@H](O)[C@@H]%11O)O[C@H](CO)[C@@H](O)[C@@H]%10O)O[C@H](CO)[C@@H](O)[C@@H]9O)O[C@H](CO)[C@@H](O)[C@@H]8O)O[C@H](CO)[C@@H](O)[C@@H]7O)O[C@H](CO)[C@@H](O)[C@@H]6O)O[C@H](CO)[C@@H](O)[C@@H]5O)O[C@H](CO)[C@@H](O)[C@@H]4O)O[C@H](CO)[C@@H](O)[C@@H]3O)O[C@H](CO)[C@@H](O)[C@@H]2O)[C@@H](O)[C@@H]1O FTSSQIKWUOOEGC-RULYVFMPSA-N 0.000 claims description 8
- 229940107187 fructooligosaccharide Drugs 0.000 claims description 8
- 238000010438 heat treatment Methods 0.000 claims description 8
- 229910052500 inorganic mineral Inorganic materials 0.000 claims description 8
- JYJIGFIDKWBXDU-MNNPPOADSA-N inulin Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)OC[C@]1(OC[C@]2(OC[C@]3(OC[C@]4(OC[C@]5(OC[C@]6(OC[C@]7(OC[C@]8(OC[C@]9(OC[C@]%10(OC[C@]%11(OC[C@]%12(OC[C@]%13(OC[C@]%14(OC[C@]%15(OC[C@]%16(OC[C@]%17(OC[C@]%18(OC[C@]%19(OC[C@]%20(OC[C@]%21(OC[C@]%22(OC[C@]%23(OC[C@]%24(OC[C@]%25(OC[C@]%26(OC[C@]%27(OC[C@]%28(OC[C@]%29(OC[C@]%30(OC[C@]%31(OC[C@]%32(OC[C@]%33(OC[C@]%34(OC[C@]%35(OC[C@]%36(O[C@@H]%37[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O%37)O)[C@H]([C@H](O)[C@@H](CO)O%36)O)[C@H]([C@H](O)[C@@H](CO)O%35)O)[C@H]([C@H](O)[C@@H](CO)O%34)O)[C@H]([C@H](O)[C@@H](CO)O%33)O)[C@H]([C@H](O)[C@@H](CO)O%32)O)[C@H]([C@H](O)[C@@H](CO)O%31)O)[C@H]([C@H](O)[C@@H](CO)O%30)O)[C@H]([C@H](O)[C@@H](CO)O%29)O)[C@H]([C@H](O)[C@@H](CO)O%28)O)[C@H]([C@H](O)[C@@H](CO)O%27)O)[C@H]([C@H](O)[C@@H](CO)O%26)O)[C@H]([C@H](O)[C@@H](CO)O%25)O)[C@H]([C@H](O)[C@@H](CO)O%24)O)[C@H]([C@H](O)[C@@H](CO)O%23)O)[C@H]([C@H](O)[C@@H](CO)O%22)O)[C@H]([C@H](O)[C@@H](CO)O%21)O)[C@H]([C@H](O)[C@@H](CO)O%20)O)[C@H]([C@H](O)[C@@H](CO)O%19)O)[C@H]([C@H](O)[C@@H](CO)O%18)O)[C@H]([C@H](O)[C@@H](CO)O%17)O)[C@H]([C@H](O)[C@@H](CO)O%16)O)[C@H]([C@H](O)[C@@H](CO)O%15)O)[C@H]([C@H](O)[C@@H](CO)O%14)O)[C@H]([C@H](O)[C@@H](CO)O%13)O)[C@H]([C@H](O)[C@@H](CO)O%12)O)[C@H]([C@H](O)[C@@H](CO)O%11)O)[C@H]([C@H](O)[C@@H](CO)O%10)O)[C@H]([C@H](O)[C@@H](CO)O9)O)[C@H]([C@H](O)[C@@H](CO)O8)O)[C@H]([C@H](O)[C@@H](CO)O7)O)[C@H]([C@H](O)[C@@H](CO)O6)O)[C@H]([C@H](O)[C@@H](CO)O5)O)[C@H]([C@H](O)[C@@H](CO)O4)O)[C@H]([C@H](O)[C@@H](CO)O3)O)[C@H]([C@H](O)[C@@H](CO)O2)O)[C@@H](O)[C@H](O)[C@@H](CO)O1 JYJIGFIDKWBXDU-MNNPPOADSA-N 0.000 claims description 8
- 229940029339 inulin Drugs 0.000 claims description 8
- 235000019421 lipase Nutrition 0.000 claims description 8
- 239000011707 mineral Substances 0.000 claims description 8
- 235000019710 soybean protein Nutrition 0.000 claims description 8
- 229940088594 vitamin Drugs 0.000 claims description 8
- 229930003231 vitamin Natural products 0.000 claims description 8
- 235000013343 vitamin Nutrition 0.000 claims description 8
- 239000011782 vitamin Substances 0.000 claims description 8
- 235000021119 whey protein Nutrition 0.000 claims description 8
- 108010004032 Bromelains Proteins 0.000 claims description 7
- 108010001682 Dextranase Proteins 0.000 claims description 7
- 239000004365 Protease Substances 0.000 claims description 7
- 235000019835 bromelain Nutrition 0.000 claims description 7
- 235000019197 fats Nutrition 0.000 claims description 7
- 150000003722 vitamin derivatives Chemical class 0.000 claims description 7
- 101710130006 Beta-glucanase Proteins 0.000 claims description 6
- 235000019871 vegetable fat Nutrition 0.000 claims description 6
- 241000167854 Bourreria succulenta Species 0.000 claims description 5
- 235000019693 cherries Nutrition 0.000 claims description 5
- BJQWBACJIAKDTJ-UHFFFAOYSA-N tetrabutylphosphanium Chemical compound CCCC[P+](CCCC)(CCCC)CCCC BJQWBACJIAKDTJ-UHFFFAOYSA-N 0.000 claims description 3
- HJHUXWBTVVFLQI-UHFFFAOYSA-N tributyl(methyl)azanium Chemical compound CCCC[N+](C)(CCCC)CCCC HJHUXWBTVVFLQI-UHFFFAOYSA-N 0.000 claims description 3
- 241000196324 Embryophyta Species 0.000 claims description 2
- -1 imide salt Chemical class 0.000 claims description 2
- 150000003949 imides Chemical class 0.000 claims description 2
- 244000267823 Hydrangea macrophylla Species 0.000 claims 3
- JRZXASDTNSFJBR-UHFFFAOYSA-L 1-ethyl-3-methylimidazol-3-ium;sulfate Chemical compound [O-]S([O-])(=O)=O.CCN1C=C[N+](C)=C1.CCN1C=C[N+](C)=C1 JRZXASDTNSFJBR-UHFFFAOYSA-L 0.000 claims 1
- 230000000694 effects Effects 0.000 abstract description 18
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- 241000699670 Mus sp. Species 0.000 description 8
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- 235000018102 proteins Nutrition 0.000 description 5
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- 241000287828 Gallus gallus Species 0.000 description 4
- 229920001503 Glucan Polymers 0.000 description 4
- 241000699666 Mus <mouse, genus> Species 0.000 description 4
- 206010057249 Phagocytosis Diseases 0.000 description 4
- 150000001875 compounds Chemical class 0.000 description 4
- 210000003743 erythrocyte Anatomy 0.000 description 4
- 210000000822 natural killer cell Anatomy 0.000 description 4
- 210000001539 phagocyte Anatomy 0.000 description 4
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- 231100000331 toxic Toxicity 0.000 description 3
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- 235000001674 Agaricus brunnescens Nutrition 0.000 description 2
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- XDZAFZVZTAGZHI-UHFFFAOYSA-N 1-ethyl-3-methyl-1,2-dihydroimidazol-1-ium;ethyl sulfate Chemical compound CCOS([O-])(=O)=O.CC[NH+]1CN(C)C=C1 XDZAFZVZTAGZHI-UHFFFAOYSA-N 0.000 description 1
- VRFOKYHDLYBVAL-UHFFFAOYSA-M 1-ethyl-3-methylimidazol-3-ium;ethyl sulfate Chemical compound CCOS([O-])(=O)=O.CCN1C=C[N+](C)=C1 VRFOKYHDLYBVAL-UHFFFAOYSA-M 0.000 description 1
- 229920002307 Dextran Polymers 0.000 description 1
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- 241000123261 Sparassidaceae Species 0.000 description 1
- 241000123241 Sparassis Species 0.000 description 1
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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
- 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
-
- 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
- A23L33/115—Fatty acids or derivatives thereof; Fats or oils
-
- 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
- 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
- A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
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- 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
- A23L33/15—Vitamins
-
- 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
- A23L33/16—Inorganic salts, minerals or trace elements
-
- 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
- A23L33/17—Amino acids, peptides or proteins
- A23L33/185—Vegetable proteins
-
- 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
- A23L33/17—Amino acids, peptides or proteins
- A23L33/19—Dairy proteins
-
- 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
-
- 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
- C08B37/0006—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid
- C08B37/0024—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid beta-D-Glucans; (beta-1,3)-D-Glucans, e.g. paramylon, coriolan, sclerotan, pachyman, callose, scleroglucan, schizophyllan, laminaran, lentinan or curdlan; (beta-1,6)-D-Glucans, e.g. pustulan; (beta-1,4)-D-Glucans; (beta-1,3)(beta-1,4)-D-Glucans, e.g. lichenan; Derivatives thereof
-
- 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
Abstract
The invention discloses a preparation method of a pharmaceutical-grade sparassis crispa polysaccharide for enhancing immunity, which comprises the following steps: (1) pretreatment of raw materials: taking a proper amount of dried sparassis crispa sporocarp, and crushing the sparassis crispa sporocarp; (2) adding ionic liquid solution into crude Sparassis crispa polysaccharide, stirring, adding enzyme for hydrolysis, filtering hydrolysate, adding ethanol into filtrate, standing, separating precipitate, and drying to obtain water-soluble Sparassis crispa polysaccharide; the invention adopts the high-efficiency specific enzymolysis extraction process of an ionic liquid/complex enzyme system to remove cellulose, fat and protein from the sparassis crispa sporocarp, thereby improving the yield of sparassis crispa polysaccharide. The viscosity of the sparassis crispa polysaccharide is reduced by adopting the green and environment-friendly ionic liquid/combined enzyme system directional hydrolysis technology, the water solubility is increased, the absorption by a human body is facilitated, and the effects of enhancing immunity, resisting tumors, reducing blood sugar and the like are remarkable.
Description
Technical Field
The invention relates to the technical field of food, in particular to a preparation method and application of pharmaceutical-grade sparassis crispa polysaccharide for enhancing immunity.
Background
Sparassis crispa is of the genus Sparassis of the order Sparassiales, family Sparassidaceae. Sparassis crispa is rich in proteins, amino acids, mineral elements and the like, is delicious in taste and rich in nutrition, is rich in active ingredients beneficial to human bodies, and has the reputation of Wan mushroom king and sunshine mushroom. The content of polysaccharide in the sparassis crispa dry product is up to 40-60%, and the sparassis crispa polysaccharide is an important active ingredient of sparassis crispa, and the sparassis crispa polysaccharide is a bioactive substance, and has multiple functions of immunoregulation, anti-tumor, anti-inflammation, antivirus, antioxidation, antiradiation, blood sugar reduction, blood fat reduction, liver protection and the like. The polysaccharide is extracted from the sparassis crispa and is prepared into various foods, medicines and the like, and the polysaccharide has wide market and economic value.
By virtue of good health-care efficacy and nutritional value of sparassis crispa, the sparassis crispa has gradually received great attention of scholars and consumers at home and abroad in recent years. The sparassis crispa polysaccharide extracted by the conventional method is difficult to industrially produce due to low yield, and is insoluble in water due to high viscosity. The problem of how to improve the yield of the Sparassis crispa polysaccharide and reduce the viscosity of the Sparassis crispa polysaccharide so as to increase the water solubility of the Sparassis crispa polysaccharide and enable the Sparassis crispa polysaccharide to be better applied to health care products and foods with special medical application is solved at present. When the sparassis crispa polysaccharide is used as a raw material to prepare health-care products and foods with special medical application, the effective absorption of the sparassis crispa polysaccharide by a human body is influenced due to the poor water solubility of the sparassis crispa polysaccharide, so that the effects of enhancing immunity, resisting tumors and reducing blood sugar are greatly reduced.
Disclosure of Invention
Aiming at the defects of the prior art, the invention provides a preparation method and application of pharmaceutical-grade sparassis crispa polysaccharide for enhancing immunity. The viscosity of the sparassis crispa polysaccharide is reduced by adopting the green and environment-friendly ionic liquid/combined enzyme system directional hydrolysis technology, the water solubility is increased, the absorption by a human body is facilitated, and the effects of enhancing immunity, resisting tumors, reducing blood sugar and the like are remarkable.
The technical scheme of the invention is as follows: a preparation method of sparassis crispa polysaccharide for enhancing immunity in pharmaceutical grade comprises the following steps:
(1) pretreatment of raw materials: taking a proper amount of dried sparassis crispa sporocarp, and crushing the sparassis crispa sporocarp; adding 10-60 times volume of ionic liquid solution, heating to 80-100 ℃, uniformly stirring to obtain mixed solution, adding 0.1-0.3 mass percent of specific endo-cellulase and lysozyme, and stirring the mixed solution for 1-2 hours at the temperature of 40-50 ℃ at the pH value of 5-7 for enzymolysis; adding lipase and bromelain with the mass percentage concentration of 0.1-0.5%, stirring the mixed solution at the temperature of 20-50 ℃ for 0.5-2 h under the condition that the pH value is 3-7, carrying out enzymolysis, carrying out centrifugal treatment on the enzymolysis solution to separate solid from liquid, discarding filtrate, and obtaining residue to obtain crude Sparassis crispa polysaccharide;
(2) taking crude Sparassis crispa polysaccharide, adding ionic liquid solution, stirring, adding enzyme for hydrolysis, filtering hydrolysate, adding ethanol into filtrate, standing, separating precipitate, and drying to obtain water-soluble Sparassis crispa polysaccharide.
Further, in the step (1), the sparassis crispa fruiting body is crushed to 800-1200 meshes by an ultrafine crusher.
Further, in the step (1), the ionic liquid solution used is an aqueous solution of 1-5% by mass of ionic liquid, wherein the ionic liquid is one or two of 1-butyl-1-methylpyrrolidine bistrifluoromethanesulfonylimide and tetrabutylphosphine tetrachloroferrite.
Further, in the step (1), the mass ratio of the specific endo-cellulase to the lysozyme is (2-5): 1, decomposing the cell wall of sparassis crispa to better dissolve intracellular glucan; the mass ratio of the lipase to the bromelain is (2-4): 1, to remove fat and protein from the raw material, thereby improving the yield of Sparassis crispa polysaccharide.
Further, in the step (2), the ionic liquid solution used is an aqueous solution of 0.5-2% by mass of an ionic liquid, wherein the ionic liquid is one or two of tributylmethylammonium bistrifluoromethylsulfonyl imide salt and 1-ethyl-3-methylimidazole ethyl sulfate.
Further, in the step (2), the enzyme is a combination enzyme of dextranase and beta-glucosaccharase, and the mass ratio of the combination enzyme is 1 (1-3), so as to degrade glycosidic bonds, and obtain the low-viscosity Sparassis crispa polysaccharide.
The ionic liquid and the enzyme preparation adopted by the invention are not limited to a specific manufacturer and can be used in the market.
The invention also aims to provide the application of the sparassis crispa polysaccharide for enhancing immunity, which is applied to a food formula with reasonable and scientific composition, has the effect of remarkably enhancing the immunity of organisms, does not have toxic or side effect and does not generate dependence.
The food formula consists of the following raw materials in parts by weight:
15-55 parts of hydrangea powder, 0-20 parts of whey protein powder, 0-20 parts of casein, 0-10 parts of soybean protein isolate, 10-50 parts of plant fat powder, 0-10 parts of fructo-oligosaccharide, 0-10 parts of inulin, 0-5 parts of acerola cherry powder, 0-5 parts of elderberry powder, 0.05-1 part of vitamin and 0.05-5 parts of mineral substances.
Further, the food formula comprises the following raw materials in parts by weight: 25-45 parts of hydrangea powder, 5-15 parts of whey protein powder, 5-15 parts of casein, 1-5 parts of soybean protein isolate, 20-40 parts of vegetable fat powder, 1-5 parts of fructo-oligosaccharide, 1-5 parts of inulin, 0.5-1.5 parts of acerola cherry powder, 0.5-2.5 parts of elderberry powder, 0.1-0.3 part of vitamin and 2-4 parts of mineral substances.
Preferably, the food formula consists of the following raw materials in parts by weight: 35 parts of hydrangea powder, 10 parts of whey protein powder, 10 parts of casein, 3 parts of soybean protein isolate, 30 parts of vegetable fat powder, 3 parts of fructo-oligosaccharide, 3.3 parts of inulin, 1 part of acerola powder, 1.5 parts of elderberry powder, 0.2 part of vitamin and 3 parts of mineral substances.
Furthermore, the hydrangea powder is prepared from the low-viscosity hydrangea polysaccharide obtained according to the technical method through a spray freeze drying technology, and is better in instant solubility.
The invention has the beneficial effects that: since most of glucan is on the cell wall of Sparassis crispa, ultramicro pulverization of Sparassis crispa fruiting body to 800-1200 mesh can mechanically destroy part of cell wall, so that glucan can be better dissolved out. The invention adopts an ionic liquid/compound enzyme system, the ionic liquid can effectively dissolve cellulose in the sparassis crispa and has good biocompatibility with enzyme, the high-efficiency enzymolysis of the sparassis crispa cell walls by specific endo-cellulase and lysozyme is promoted, the dissolution rate of intracellular glucan is improved, and the aim of improving the yield of the sparassis crispa polysaccharide is fulfilled. Because an ionic liquid/combined enzyme system is adopted, the dextran enzyme and the beta-glucanase combined enzyme can efficiently directionally control and degrade the macromolecular glycosidic chain segment in the ionic liquid, and the low-viscosity Sparassis crispa polysaccharide is obtained. The Sparassis crispa polysaccharide prepared by the method is prepared by a latest spray freeze drying technology, so that loss of bioactive substances is reduced, the structure of the product is loose and porous, the instant solubility of the product is improved, the absorption rate of a human body is improved, and the immunity of the human body is enhanced. The invention has reasonable and scientific application formula composition by taking low-viscosity Sparassis crispa polysaccharide as a raw material, has the function of obviously enhancing the immunity of the organism, has no toxic or side effect, and does not generate dependence.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products available commercially.
Example 1
The preparation method comprises the following steps of preparing pharmaceutical-grade sparassis crispa polysaccharide for enhancing immunity:
1. pretreating the raw materials by an ionic liquid/compound enzyme system:
taking 1kg of dried sparassis crispa fruiting body, and crushing the sparassis crispa fruiting body into 800 meshes by using an ultrafine crusher; adding 10L of 1 wt% 1-butyl-1-methylpyrrolidine bistrifluoromethanesulfonylimide ionic liquid solution, heating to 85 ℃, uniformly stirring to obtain a mixed solution, cooling to 45 ℃, adjusting the pH to 6, and adding a complex enzyme with the mass percentage concentration of 0.2%, wherein the complex enzyme comprises specific endo-cellulase in parts by weight: and (2) 1:1, stirring the mixed solution at 45 ℃ for 1.5h for enzymolysis, and then adding 0.3% of complex enzyme in percentage by weight, wherein the complex enzyme comprises the following components in parts by weight: and (3) mixing the mixed solution at 40 ℃ for 1.5h to carry out enzymolysis, carrying out centrifugal treatment on the enzymolysis solution to separate solid from liquid, discarding filtrate to obtain residue, carrying out repeated enzymolysis and extraction on the solid residue for 3 times, and combining the obtained residues to obtain the crude polysaccharide of the Sparassis crispa.
In the steps, the cell wall structure is broken through the ultrafine grinder to promote the polysaccharide content to be separated out, the cell wall structure is further hydrolyzed in an ionic liquid solution through specific endo-cellulase and lysozyme in the complex enzyme, and fat and protein contained in the Sparassis crispa are hydrolyzed through lipase and bromelain, so that the yield and the purity of the Sparassis crispa polysaccharide are improved.
2. Preparing low-viscosity Sparassis crispa polysaccharide liquid by enzymolysis of an ionic liquid/combined enzyme system:
adding 15 times volume of 0.8 wt% of tributyl methyl ammonium bis (trifluoromethyl) sulfonyl imide salt ionic liquid solution into the residue Sparassis crispa crude polysaccharide obtained in the step 1, heating to 85 ℃, uniformly stirring to obtain a mixed solution, cooling to 50 ℃, adjusting the pH to 5, adding a combined enzyme with the mass percent concentration of 0.07%, wherein the combined enzyme comprises dextranase and beta-glucanase 1:1 in parts by weight, uniformly stirring to obtain a mixed solution, stirring the mixed solution at 50 ℃ for enzymolysis for 3 hours, centrifuging an enzymolysis solution in a centrifuge (the rotation speed is 4000r/min) for 15min, separating solid from liquid, discarding residues, and obtaining a filtrate to obtain a low-viscosity Sparassis crispa polysaccharide liquid.
3. Concentrating and precipitating with ethanol:
concentrating the low-viscosity Sparassis crispa polysaccharide liquid obtained in the step (2), inactivating at 90 ℃ for 10min, and cooling to room temperature; adding 95% ethanol solution with 3 times volume, standing, and filtering to obtain precipitate.
4. Freezing and spray drying:
and (3) drying the precipitate obtained in the step (3) in a freezing spray dryer, and crushing to 60 meshes to obtain the final Sparassis crispa polysaccharide powder product 1.
Example 2
The preparation method comprises the following steps of preparing pharmaceutical-grade sparassis crispa polysaccharide for enhancing immunity:
1. pretreating the raw materials by an ionic liquid/compound enzyme system:
taking 1kg of dried sparassis crispa fruiting body, and crushing the sparassis crispa fruiting body into 1000 meshes by using an ultrafine crusher; adding 15L of 1 wt% tetrabutyl phosphine tetrachloroferrite ionic liquid solution, heating to 85 ℃, uniformly stirring to obtain a mixed solution, cooling to 45 ℃, regulating the pH to 6, and adding a complex enzyme with the mass percent concentration of 0.3%, wherein the complex enzyme comprises the following specific endo-cellulase in parts by weight: and (2) mixing the lysozyme with the mixed solution at the temperature of 45 ℃ for 1.5h for enzymolysis, and then adding a complex enzyme with the mass percentage concentration of 0.3%, wherein the complex enzyme comprises the following components in parts by weight: and (3) stirring the mixed solution at the temperature of 40 ℃ for 1.5h to carry out enzymolysis, centrifuging the enzymolysis solution to separate solid from liquid, discarding filtrate to obtain residue, repeatedly carrying out enzymolysis and leaching on the solid residue for 3 times, and combining the obtained residues to obtain the crude polysaccharide of the Sparassis crispa.
In the steps, the cell wall structure is broken through the ultrafine grinder to promote the polysaccharide content to be separated out, the cell wall structure is further hydrolyzed in an ionic liquid solution through specific endo-cellulase and lysozyme in the complex enzyme, and fat and protein contained in the Sparassis crispa are hydrolyzed through lipase and bromelain, so that the yield and the purity of the Sparassis crispa polysaccharide are improved.
2. Preparing low-viscosity Sparassis crispa polysaccharide liquid by enzymolysis of an ionic liquid/combined enzyme system:
adding 15 times volume of 0.6 wt% 1-ethyl-3-methylimidazolium ethyl sulfate ionic liquid solution into the residue Sparassis crispa crude polysaccharide obtained in the step 1, heating to 85 ℃, uniformly stirring to obtain a mixed solution, cooling to 50 ℃, adjusting the pH to 5, adding a combined enzyme with the mass percent concentration of 0.07%, wherein the combined enzyme comprises dextranase and beta-glucanase which are 1:2 in parts by weight, uniformly stirring to obtain a mixed solution, stirring the mixed solution at 50 ℃ for enzymolysis for 3 hours, centrifuging the enzymolysis solution in a centrifuge (the rotation speed is 4000r/min) for 15min, separating solid from liquid, discarding residues, and obtaining a filtrate to obtain a low-viscosity Sparassis crispa polysaccharide liquid.
3. Concentrating and precipitating with ethanol:
concentrating the low-viscosity Sparassis crispa polysaccharide liquid obtained in the step (2), inactivating at 90 ℃ for 10min, and cooling to room temperature; adding 95% ethanol solution with 3 times volume, standing, and filtering to obtain precipitate.
4. Freezing and spray drying:
and (3) drying the precipitate obtained in the step (3) in a freezing spray dryer, and crushing to 60 meshes to obtain a final Sparassis crispa polysaccharide powder product 2.
Example 3
The preparation method comprises the following steps of preparing pharmaceutical-grade sparassis crispa polysaccharide for enhancing immunity:
1. pretreating the raw materials by an ionic liquid/compound enzyme system:
taking 1kg of dried sparassis crispa fruiting body, and crushing the sparassis crispa fruiting body into 1200 meshes by using an ultrafine crusher; adding 10L of 1.5 wt% 1-butyl-1-methylpyrrolidine bistrifluoromethanesulfonylimide ionic liquid solution, heating to 85 ℃, uniformly stirring to obtain a mixed solution, cooling to 45 ℃, adjusting the pH to 6, and adding a complex enzyme with the mass percentage concentration of 0.2%, wherein the complex enzyme comprises specific endo-cellulase in parts by weight: and (2) mixing lysozyme with the ratio of 4:1, stirring the mixed solution at the temperature of 45 ℃ for 1.5h for enzymolysis, and then adding a complex enzyme with the mass percentage concentration of 0.3%, wherein the complex enzyme comprises the following lipase in parts by weight: and (3) stirring the mixed solution at the temperature of 40 ℃ for 1.5h to carry out enzymolysis, centrifuging the enzymolysis solution to separate solid from liquid, discarding filtrate to obtain residue, repeatedly carrying out enzymolysis and leaching on the solid residue for 3 times, and combining the obtained residues to obtain the crude polysaccharide of the Sparassis crispa.
In the steps, the cell wall structure is broken through the ultrafine grinder to promote the polysaccharide content to be separated out, the cell wall structure is further hydrolyzed in an ionic liquid solution through specific endo-cellulase and lysozyme in the complex enzyme, and fat and protein contained in the Sparassis crispa are hydrolyzed through lipase and bromelain, so that the yield and the purity of the Sparassis crispa polysaccharide are improved.
2. Preparing low-viscosity Sparassis crispa polysaccharide liquid by enzymolysis of an ionic liquid/combined enzyme system:
adding 15 times volume of 0.7 wt% of tributyl methyl ammonium bistrifluoromethylsulfonyl imide ionic liquid solution into the residue Sparassis crispa crude polysaccharide obtained in the step 1, heating to 85 ℃, uniformly stirring to obtain a mixed solution, cooling to 50 ℃, adjusting the pH to 5, adding a combined enzyme with the mass percentage concentration of 0.07%, wherein the combined enzyme comprises beta-dextranase and beta-glucosaccharase in parts by weight, uniformly stirring to obtain a mixed solution, stirring the mixed solution at 50 ℃ for enzymolysis for 3h, centrifuging the enzymolysis solution in a centrifuge (the rotation speed is 4000r/min) for 15min, separating solid from liquid, discarding residues, and obtaining a filtrate to obtain a low-viscosity Sparassis crispa polysaccharide liquid.
3. Concentrating and precipitating with ethanol:
concentrating the low-viscosity Sparassis crispa polysaccharide liquid obtained in the step (2), inactivating at 90 ℃ for 10min, and cooling to room temperature; adding 95% ethanol solution with 3 times volume, standing, and filtering to obtain precipitate.
4. Freezing and spray drying:
and (3) drying the precipitate obtained in the step (3) in a freezing spray dryer, and crushing to 60 meshes to obtain a final Sparassis crispa polysaccharide powder product 3.
Comparative example 1
The difference from example 1 is that the fruit body of Sparassis crispa was pulverized to 200 mesh, and the rest of the procedure was not changed.
Comparative example 2
In contrast to example 1, 10L of a 1% wt 1-butyl-1-methylpyrrolidine bistrifluoromethanesulfonylimide ionic liquid solution was replaced in step 1 with 10L of distilled water, and the rest of the procedure was unchanged.
Comparative example 3
Unlike example 1, the enzyme added in step 1 was a single specific endo-cellulase, and the other processes were not changed.
Comparative example 4
The difference from example 1 is that the enzyme added in step 1 is a single lysozyme and the other processes are not changed.
Comparative example 5
Different from the example 1, 15 times of volume of the ionic liquid solution of 0.8 percent weight of tributyl methyl ammonium bis (trifluoromethyl) sulfonyl imide salt is replaced by 15 times of volume of distilled water in the step 2, and other processes are not changed,
comparative example 6
Different from the example 2, the enzyme added in the step 3 is single dextranase, other processes are not changed, and the final sparassis crispa polysaccharide powder product 4 is obtained.
Comparative example 7
Different from the example 2, the enzyme added in the step 3 is single beta-glucosaccharase, other processes are not changed, and the final Sparassis crispa polysaccharide powder product 5 is obtained.
The crude sparassis crispa polysaccharides obtained in step 1 of the above examples 1, 2 and 3 and comparative examples 1, 2, 3 and 4 were respectively weighed, and the respective yields were calculated, and the data obtained are shown in table 1 below:
TABLE 1
As can be seen from the data in Table 1, the comparison between example 1 and comparative example 1 shows that the finer the mesh number of the crushed Sparassis crispa fruiting body is, the higher the yield of Sparassis crispa polysaccharide product is, which means that the ultra-micro mechanical crushing damages the cell wall of Sparassis crispa fruiting body more, so that the polysaccharide is dissolved out more; comparing example 1 with comparative example 2, it can be seen that the yield of the polysaccharide product of sparassis crispa is much higher in the ionic liquid/complex enzyme system than in the pure water/complex enzyme system; the comparison between example 1 and comparative examples 3 and 4 shows that the complex enzyme has a remarkable effect of damaging the cell walls of the sparassis crispa fruiting body compared with a single enzyme.
The final sparassis crispa polysaccharide powder products obtained in the above examples 1, 2 and 3 and comparative examples 5, 6 and 7 were prepared into aqueous solutions with mass percentage concentrations of 0.1%, 0.2%, 0.5% and 1.0%, respectively, and the viscosity was measured by a rotational viscometer, and the data obtained are shown in the following table 2:
TABLE 2
As can be seen from the data in Table 2, when the mass percentage concentrations of the aqueous solutions are the same, the comparison between the example 1 and the comparative example 5 shows that the viscosity of the Sparassis crispa polysaccharide product prepared by the ionic liquid/combined enzyme system is greatly reduced compared with that prepared by pure water/combined enzyme system, and the liquid/combined enzyme system is more beneficial to the enzymolysis reaction of Sparassis crispa polysaccharide; the viscosity of the Sparassis crispa polysaccharide products prepared in examples 1, 2 and 3 is much lower than that of the Sparassis crispa polysaccharide products prepared in comparative examples 6 and 7, which shows that the hydrolysis effect of the combined enzyme of dextranase and beta-glucanase is better than that of a single enzyme, the product viscosity is low, and the molecular weight is lower, so that the product is easier to absorb by human bodies.
The final Sparassis crispa polysaccharide powder products 1, 2, 3 and 4, 5 obtained in the above examples 1, 2, 3 and comparative examples 6, 7 were each prepared as an aqueous solution having a mass percentage concentration of 0.2%, and the state of the aqueous solution was observed.
The aqueous solutions of the Sparassis crispa polysaccharide products 1, 2 and 3 obtained in examples 1, 2 and 3 were found to be highly transparent, clear and transparent, whereas the aqueous solution of the Sparassis crispa polysaccharide product 4 obtained in comparative example 6 had a small amount of suspended flocs, and the aqueous solution of the Sparassis crispa polysaccharide product 5 obtained in comparative example 7 had not very good transparency, but was not clear and transparent, further showing that the Sparassis crispa polysaccharide product obtained by hydrolysis with a combination of dextranase and beta-glucanase had good water solubility, clear and transparent appearance, and was more easily favored by the market.
Example 8
An application formula of sparassis crispa polysaccharide for enhancing immunity comprises the following raw materials in parts by weight:
35 parts of hydrangea powder, 10 parts of whey protein powder, 10 parts of casein, 3 parts of soybean protein isolate, 30 parts of vegetable fat powder, 3 parts of fructo-oligosaccharide, 3.3 parts of inulin, 1 part of acerola powder, 1.5 parts of elderberry powder, 0.2 part of vitamin and 3 parts of mineral substances.
The raw material Sparassis crispa powder is Sparassis crispa polysaccharide powder product 2 prepared in example 2, and other raw materials are all sold in the conventional market. The formula is comprehensive and balanced in nutrition, and the components synergize to achieve the effect of improving the immunity of the organism.
The preparation method comprises the following steps: the preparation method comprises the steps of premixing vitamins and minerals uniformly, adding acerola cherry powder and elderberry powder, mixing uniformly, adding soybean protein isolate, fructo-oligosaccharide and inulin, mixing uniformly, adding whey protein powder and casein, mixing uniformly, adding hydrangea powder and vegetable fat powder, and mixing uniformly to obtain the application formula product M of the immunity-enhancing hydrangea polysaccharide.
< evaluation test of Immunity-enhancing Effect >
Female mice of 6 weeks of age were selected for the experiment and divided into 4 groups by body weight. The human recommended dose of the sparassis crispa polysaccharide application formula product M is 0.5g/kg/d, and the equivalent dose of a mouse is 10 times of the human recommended dose. The human body recommended dose is 5 times, 10 times and 30 times respectively as low, medium and high dose groups. The gavage method is adopted, the gavage is performed once a day, and the control group is filled with distilled water. After each group of mice continuously takes the product M for 30 days, the mice freely eat and drink water, the proper temperature (25 +/-1.0 ℃) and humidity and silence of the feeding environment are ensured, and all indexes are measured.
On day 30 of administration, body weight was weighed. The mice are sacrificed, the spleen and the thymus are taken and weighed, and the organ coefficient is calculated, wherein the ratio of the spleen (thymus) to the spleen (thymus) is weight/weight; the toe swelling degree was also measured, and the results are shown in Table 3.
TABLE 3 Effect of product M on mouse thymus, spleen quality and toe swelling
As can be seen from Table 3, after the product M of the present invention is orally administered for 30 days, the thymus ratio and the spleen/body weight ratio of each dose group are not significantly different (p >0.05) compared with the blank control group after the mice are administered with different doses of the test substance for 30 days, which indicates that the spleen and the thymus can normally exert their physiological functions without causing adverse stimulation to the spleen and the thymus when the product M of the present invention is administered. Meanwhile, after the product M is taken, the toe swelling difference values of the low, medium and high dose groups are obviously different from those of a blank control group (p is less than 0.05), which shows that the product M can obviously improve the cellular immunity of the organism.
Further, on day 30 of administration, 1ml of 20% (V/V) suspension of chicken red blood cells was intraperitoneally injected into each mouse, the abdominal cavity was dropped into a tablet, incubated at 37 ℃ for 30min, fixed, stained, and then macrophages were counted under a microscope, and the phagocytosis index and phagocytosis rate were calculated. Phagocytosis index is the total number of chicken red blood cells phagocytosed per number of phagocytic cells counted; the phagocytosis rate is the number of phagocytes that phagocytose chicken erythrocytes/the number of phagocytes counted × 100%, and the results are shown in table 4.
TABLE 4 Effect of product M on phagocytic function of mice
The phagocytic function of the macrophages can be judged by the total number of phagocytic cells of the abdominal cavity injection chicken erythrocytes of the mice. As can be seen from table 4, the phagocytic index and rate were significantly higher in the low, medium and high dose groups than in the blank control group (p <0.05) after the oral administration of the product M for 30 days.
Further, on day 30 of the administration, the mouse NK cell activity was examined, and the results are shown in Table 5.
TABLE 5 Effect of product M on NK cell Activity in mice
As can be seen from Table 5, the NK cell activities of the low, medium and high dose groups were significantly different (p <0.05) from those of the blank control group after administration of the product M of the present invention, indicating that the product M has a promoting effect on NK cell activity.
In conclusion, the application formula of the sparassis crispa polysaccharide for enhancing immunity provided by the invention can remarkably enhance the immunity of organisms; the proportion of the raw materials is reasonable, and the maximum synergistic effect can be exerted under the proportion; meanwhile, the composition has a simple formula, so the composition has no toxic or side effect, and patients are not easy to generate dependence. The preparation method of the composition is simple, and various parameters are controllable, so that the composition can be produced in a large scale.
The above are merely characteristic embodiments of the present invention, and do not limit the scope of the present invention in any way. All technical solutions formed by equivalent exchanges or equivalent substitutions fall within the protection scope of the present invention.
Claims (9)
1. A preparation method of sparassis crispa polysaccharide for enhancing immunity in pharmaceutical grade is characterized by comprising the following steps:
(1) pretreatment of raw materials: taking a proper amount of dried sparassis crispa sporocarp, and crushing the sparassis crispa sporocarp; adding 10-60 times volume of ionic liquid solution, heating to 80-100 ℃, uniformly stirring to obtain mixed solution, adding 0.1-0.3 mass percent of specific endo-cellulase and lysozyme, and stirring the mixed solution for 1-2 hours at the temperature of 40-50 ℃ under the condition that the pH is =5-7 for enzymolysis; adding lipase and bromelain with the mass percentage concentration of 0.1-0.5%, stirring the mixed solution at the temperature of 20-50 ℃ for 0.5-2 h under the condition of pH =3-7 for enzymolysis, centrifuging the enzymolysis solution to separate solid from liquid, and removing filtrate to obtain residue to obtain crude Sparassis crispa polysaccharide;
(2) taking crude Sparassis crispa polysaccharide, adding ionic liquid solution, stirring, adding enzyme for hydrolysis, filtering hydrolysate, adding ethanol into filtrate, standing, separating precipitate, and drying to obtain water-soluble Sparassis crispa polysaccharide.
2. The method for preparing sparassis crispa polysaccharide for pharmaceutical grade boosting immunity as claimed in claim 1, wherein in step (1), sparassis crispa fruiting body is pulverized to 800-1200 mesh by ultramicro pulverizer.
3. The method for preparing sparassis crispa polysaccharide for pharmaceutical grade boosting immunity according to claim 1, wherein in the step (1), the ionic liquid solution is 1-5% by mass of an aqueous solution of ionic liquid, wherein the ionic liquid is one or two of 1-butyl-1-methylpyrrolidine bistrifluoromethylsulfonyl imide and tetrabutyl phosphine tetrachloroferrite.
4. The method for preparing sparassis crispa polysaccharide for pharmaceutical grade boosting immunity according to claim 1, wherein in the step (1), the mass ratio of the specific endo-cellulase to the lysozyme is (2-5): 1, the mass ratio of the lipase to the bromelain is (2-4): 1.
5. the method for preparing sparassis crispa polysaccharide for pharmaceutical grade boosting immunity according to claim 1, wherein in the step (2), the ionic liquid solution is 0.5-2% by mass of an aqueous solution of an ionic liquid, wherein the ionic liquid is one or two of tributylmethylammonium bistrifluoromethylsulfonyl imide salt and 1-ethyl-3-methylimidazolium sulfate.
6. The method for preparing sparassis crispa polysaccharide for pharmaceutical grade boosting immunity according to claim 1, wherein in the step (2), the enzyme is a combination of dextranase and beta-glucanase, and the mass ratio is 1 (1-3).
7. The application of sparassis crispa polysaccharide for enhancing immunity is characterized by being applied to the following food formula in parts by weight:
15-55 parts of hydrangea powder, 0-20 parts of whey protein powder, 0-20 parts of casein, 0-10 parts of soybean protein isolate, 10-50 parts of plant fat powder, 0-10 parts of fructo-oligosaccharide, 0-10 parts of inulin, 0-5 parts of acerola cherry powder, 0-5 parts of elderberry powder, 0.05-1 part of vitamin and 0.05-5 parts of mineral substances.
8. The application of sparassis crispa polysaccharide for enhancing immunity as claimed in claim 7, wherein the food formula comprises the following raw material formula in parts by weight: 25-45 parts of hydrangea powder, 5-15 parts of whey protein powder, 5-15 parts of casein, 1-5 parts of soybean protein isolate, 20-40 parts of vegetable fat powder, 1-5 parts of fructo-oligosaccharide, 1-5 parts of inulin, 0.5-1.5 parts of acerola cherry powder, 0.5-2.5 parts of elderberry powder, 0.1-0.3 part of vitamin and 2-4 parts of mineral substances.
9. The application of sparassis crispa polysaccharide for enhancing immunity as claimed in claim 7, wherein the food formula comprises the following raw material formula in parts by weight: 35 parts of hydrangea powder, 10 parts of whey protein powder, 10 parts of casein, 3 parts of soybean protein isolate, 30 parts of vegetable fat powder, 3 parts of fructo-oligosaccharide, 3.3 parts of inulin, 1 part of acerola powder, 1.5 parts of elderberry powder, 0.2 part of vitamin and 3 parts of mineral substances.
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| CN113402625A (en) * | 2021-06-28 | 2021-09-17 | 安发（福建）生物科技有限公司 | Inonotus obliquus polysaccharide and extraction method thereof |
Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102277400A (en) * | 2011-08-15 | 2011-12-14 | 上海科爱生物技术有限公司 | Extract containing beta-1,3-D-glucan and use thereof |
| CN103113487A (en) * | 2013-02-21 | 2013-05-22 | 嘉兴职业技术学院 | Method for preparing flammulina velutipes polysaccharides and protein efficiently synchronously |
| CN104146238A (en) * | 2014-07-11 | 2014-11-19 | 吉林大学 | Health food with function of enhancing immunity of human body |
| CN106434373A (en) * | 2016-09-29 | 2017-02-22 | 宁波希诺亚海洋生物科技有限公司 | High-density fermentation medium formula of sparassis crispa and pharmaceutical grade glucan preparation method of high-density fermentation medium formula |
| CN108324742A (en) * | 2018-03-20 | 2018-07-27 | 福建容益菌业科技研发有限公司 | Sparassis crispa electuary and preparation method thereof |
| CN110606899A (en) * | 2019-10-09 | 2019-12-24 | 深圳市清生元生物技术股份有限公司 | Method for extracting Sparassis crispa polysaccharide by enzymolysis |
| CN110776582A (en) * | 2019-11-25 | 2020-02-11 | 福清市火麒麟食用菌技术开发有限公司 | Method for extracting β -glucan in sparassis crispa |
| CN111990461A (en) * | 2020-08-18 | 2020-11-27 | 北安宜品努卡乳业有限公司 | Mixed milk powder for enhancing immunity and preparation method thereof |
-
2020
- 2020-12-25 CN CN202011561116.4A patent/CN112708648A/en active Pending
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102277400A (en) * | 2011-08-15 | 2011-12-14 | 上海科爱生物技术有限公司 | Extract containing beta-1,3-D-glucan and use thereof |
| CN103113487A (en) * | 2013-02-21 | 2013-05-22 | 嘉兴职业技术学院 | Method for preparing flammulina velutipes polysaccharides and protein efficiently synchronously |
| CN104146238A (en) * | 2014-07-11 | 2014-11-19 | 吉林大学 | Health food with function of enhancing immunity of human body |
| CN106434373A (en) * | 2016-09-29 | 2017-02-22 | 宁波希诺亚海洋生物科技有限公司 | High-density fermentation medium formula of sparassis crispa and pharmaceutical grade glucan preparation method of high-density fermentation medium formula |
| CN108324742A (en) * | 2018-03-20 | 2018-07-27 | 福建容益菌业科技研发有限公司 | Sparassis crispa electuary and preparation method thereof |
| CN110606899A (en) * | 2019-10-09 | 2019-12-24 | 深圳市清生元生物技术股份有限公司 | Method for extracting Sparassis crispa polysaccharide by enzymolysis |
| CN110776582A (en) * | 2019-11-25 | 2020-02-11 | 福清市火麒麟食用菌技术开发有限公司 | Method for extracting β -glucan in sparassis crispa |
| CN111990461A (en) * | 2020-08-18 | 2020-11-27 | 北安宜品努卡乳业有限公司 | Mixed milk powder for enhancing immunity and preparation method thereof |
Non-Patent Citations (7)
| Title |
|---|
| 付丽: "《食品营养与卫生》", 31 January 2013, 中国轻工业出版社 * |
| 徐明、魏建英、虞璧丹: "干粉吸入剂的制粒技术研究进展", 《中国现代中药》 * |
| 杜祯等: "喷雾冷冻干燥技术进展及其在药剂学中的应用", 《中国药学杂志》 * |
| 洪小君: "绣球菌多糖提取、分离纯化及生物活性研究", 《中国优秀硕士学位论文全文数据库 工程科技Ⅰ辑》 * |
| 相萍萍等: "离子液体在多糖溶解中的应用进展", 《食品工业科技》 * |
| 程俊文等: "香菇子实体多糖分步酶解法提取研究", 《食用菌学报》 * |
| 阚建全: "《食品化学》", 30 September 2008, 中国农业大学出版社 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113402625A (en) * | 2021-06-28 | 2021-09-17 | 安发（福建）生物科技有限公司 | Inonotus obliquus polysaccharide and extraction method thereof |
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