CN108976314A - A kind of ganoderma lucidum beta glucan and preparation method thereof and preparing the application in immunoregulation medicament - Google Patents
A kind of ganoderma lucidum beta glucan and preparation method thereof and preparing the application in immunoregulation medicament Download PDFInfo
- Publication number
- CN108976314A CN108976314A CN201810426702.4A CN201810426702A CN108976314A CN 108976314 A CN108976314 A CN 108976314A CN 201810426702 A CN201810426702 A CN 201810426702A CN 108976314 A CN108976314 A CN 108976314A
- Authority
- CN
- China
- Prior art keywords
- beta glucan
- ganoderma lucidum
- water
- preparation
- lucidum beta
- 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
- 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 69
- 229920002498 Beta-glucan Polymers 0.000 title claims abstract description 69
- 240000008397 Ganoderma lucidum Species 0.000 title claims abstract description 58
- 235000001637 Ganoderma lucidum Nutrition 0.000 title claims abstract description 58
- 238000002360 preparation method Methods 0.000 title claims abstract description 19
- 239000003814 drug Substances 0.000 title claims abstract description 15
- 230000007365 immunoregulation Effects 0.000 title claims abstract description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 29
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims abstract description 26
- 150000004676 glycans Chemical class 0.000 claims abstract description 19
- 229920001282 polysaccharide Polymers 0.000 claims abstract description 18
- 239000005017 polysaccharide Substances 0.000 claims abstract description 18
- 239000000284 extract Substances 0.000 claims abstract description 13
- 150000002772 monosaccharides Chemical group 0.000 claims abstract description 12
- KVYRCBOUKXJXDK-UHFFFAOYSA-N 3,4-dimethylphenazine-1,2-diamine hydrochloride Chemical compound Cl.C1=CC=CC2=NC3=C(C)C(C)=C(N)C(N)=C3N=C21 KVYRCBOUKXJXDK-UHFFFAOYSA-N 0.000 claims abstract description 11
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims abstract description 10
- 230000004957 immunoregulator effect Effects 0.000 claims abstract description 10
- 235000009508 confectionery Nutrition 0.000 claims abstract description 9
- 229940079593 drug Drugs 0.000 claims abstract description 9
- 239000000463 material Substances 0.000 claims abstract description 9
- 238000002474 experimental method Methods 0.000 claims abstract description 8
- 238000012869 ethanol precipitation Methods 0.000 claims abstract description 7
- AEMOLEFTQBMNLQ-AQKNRBDQSA-N D-glucopyranuronic acid Chemical compound OC1O[C@H](C(O)=O)[C@@H](O)[C@H](O)[C@H]1O AEMOLEFTQBMNLQ-AQKNRBDQSA-N 0.000 claims abstract description 4
- IAJILQKETJEXLJ-UHFFFAOYSA-N Galacturonsaeure Natural products O=CC(O)C(O)C(O)C(O)C(O)=O IAJILQKETJEXLJ-UHFFFAOYSA-N 0.000 claims abstract description 4
- 239000003957 anion exchange resin Substances 0.000 claims abstract description 4
- 229940097043 glucuronic acid Drugs 0.000 claims abstract description 4
- HEMHJVSKTPXQMS-UHFFFAOYSA-M sodium hydroxide Inorganic materials [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 20
- 239000000243 solution Substances 0.000 claims description 19
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 12
- 238000000605 extraction Methods 0.000 claims description 12
- 238000000034 method Methods 0.000 claims description 11
- 239000006228 supernatant Substances 0.000 claims description 11
- 241000222336 Ganoderma Species 0.000 claims description 10
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 8
- 239000007864 aqueous solution Substances 0.000 claims description 8
- 239000000287 crude extract Substances 0.000 claims description 8
- 239000012153 distilled water Substances 0.000 claims description 8
- 239000007788 liquid Substances 0.000 claims description 8
- 229920001503 Glucan Polymers 0.000 claims description 6
- KWYUFKZDYYNOTN-UHFFFAOYSA-M Potassium hydroxide Chemical compound [OH-].[K+] KWYUFKZDYYNOTN-UHFFFAOYSA-M 0.000 claims description 6
- 239000011780 sodium chloride Substances 0.000 claims description 6
- 235000002639 sodium chloride Nutrition 0.000 claims description 6
- 239000002253 acid Substances 0.000 claims description 5
- 238000010438 heat treatment Methods 0.000 claims description 5
- 229960002668 sodium chloride Drugs 0.000 claims description 5
- GUBGYTABKSRVRQ-WFVLMXAXSA-N DEAE-cellulose Chemical compound OC1C(O)C(O)C(CO)O[C@H]1O[C@@H]1C(CO)OC(O)C(O)C1O GUBGYTABKSRVRQ-WFVLMXAXSA-N 0.000 claims description 4
- 229920005654 Sephadex Polymers 0.000 claims description 4
- 239000012507 Sephadex™ Substances 0.000 claims description 4
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 claims description 4
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 claims description 4
- 235000019441 ethanol Nutrition 0.000 claims description 4
- BWHMMNNQKKPAPP-UHFFFAOYSA-L potassium carbonate Chemical compound [K+].[K+].[O-]C([O-])=O BWHMMNNQKKPAPP-UHFFFAOYSA-L 0.000 claims description 4
- 239000002994 raw material Substances 0.000 claims description 4
- 239000003513 alkali Substances 0.000 claims description 3
- 239000012043 crude product Substances 0.000 claims description 3
- 238000004108 freeze drying Methods 0.000 claims description 3
- CTYRPMDGLDAWRQ-UHFFFAOYSA-N phenyl hydrogen sulfate Chemical compound OS(=O)(=O)OC1=CC=CC=C1 CTYRPMDGLDAWRQ-UHFFFAOYSA-N 0.000 claims description 3
- 230000001376 precipitating effect Effects 0.000 claims description 3
- 238000000746 purification Methods 0.000 claims description 3
- 239000012609 strong anion exchange resin Substances 0.000 claims description 3
- 229920000936 Agarose Polymers 0.000 claims description 2
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 claims description 2
- 239000000908 ammonium hydroxide Substances 0.000 claims description 2
- 230000018044 dehydration Effects 0.000 claims description 2
- 238000006297 dehydration reaction Methods 0.000 claims description 2
- 238000001914 filtration Methods 0.000 claims description 2
- 229910000027 potassium carbonate Inorganic materials 0.000 claims description 2
- 238000007670 refining Methods 0.000 claims description 2
- 239000013049 sediment Substances 0.000 claims description 2
- 229910000030 sodium bicarbonate Inorganic materials 0.000 claims description 2
- 235000017557 sodium bicarbonate Nutrition 0.000 claims description 2
- 229910000029 sodium carbonate Inorganic materials 0.000 claims description 2
- 239000007787 solid Substances 0.000 claims description 2
- WMFOQBRAJBCJND-UHFFFAOYSA-M Lithium hydroxide Chemical compound [Li+].[OH-] WMFOQBRAJBCJND-UHFFFAOYSA-M 0.000 claims 3
- 230000001900 immune effect Effects 0.000 claims 1
- 239000008103 glucose Substances 0.000 abstract description 13
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 abstract description 4
- 238000005516 engineering process Methods 0.000 abstract description 2
- 239000000203 mixture Substances 0.000 description 7
- 206010057249 Phagocytosis Diseases 0.000 description 6
- 230000008782 phagocytosis Effects 0.000 description 6
- 238000004458 analytical method Methods 0.000 description 5
- 238000002835 absorbance Methods 0.000 description 4
- 238000005259 measurement Methods 0.000 description 4
- 108090000790 Enzymes Proteins 0.000 description 3
- 102000004190 Enzymes Human genes 0.000 description 3
- 238000003556 assay Methods 0.000 description 3
- 238000001977 collision-induced dissociation tandem mass spectrometry Methods 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 229940088598 enzyme Drugs 0.000 description 3
- 239000012634 fragment Substances 0.000 description 3
- 238000001228 spectrum Methods 0.000 description 3
- SRBFZHDQGSBBOR-IOVATXLUSA-N D-xylopyranose Chemical compound O[C@@H]1COC(O)[C@H](O)[C@H]1O SRBFZHDQGSBBOR-IOVATXLUSA-N 0.000 description 2
- XLYOFNOQVPJJNP-ZSJDYOACSA-N Heavy water Chemical group [2H]O[2H] XLYOFNOQVPJJNP-ZSJDYOACSA-N 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 150000001720 carbohydrates Chemical class 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 238000012512 characterization method Methods 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 238000006073 displacement reaction Methods 0.000 description 2
- 238000005227 gel permeation chromatography Methods 0.000 description 2
- 125000002791 glucosyl group Chemical group C1([C@H](O)[C@@H](O)[C@H](O)[C@H](O1)CO)* 0.000 description 2
- 238000004192 high performance gel permeation chromatography Methods 0.000 description 2
- 238000006386 neutralization reaction Methods 0.000 description 2
- 239000013641 positive control Substances 0.000 description 2
- 238000001556 precipitation Methods 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- NWUYHJFMYQTDRP-UHFFFAOYSA-N 1,2-bis(ethenyl)benzene;1-ethenyl-2-ethylbenzene;styrene Chemical compound C=CC1=CC=CC=C1.CCC1=CC=CC=C1C=C.C=CC1=CC=CC=C1C=C NWUYHJFMYQTDRP-UHFFFAOYSA-N 0.000 description 1
- 238000005160 1H NMR spectroscopy Methods 0.000 description 1
- 238000001026 1H--1H correlation spectroscopy Methods 0.000 description 1
- 238000004701 1H-13C HSQC Methods 0.000 description 1
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical class CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 1
- 108010059892 Cellulase Proteins 0.000 description 1
- SHZGCJCMOBCMKK-UHFFFAOYSA-N D-mannomethylose Natural products CC1OC(O)C(O)C(O)C1O SHZGCJCMOBCMKK-UHFFFAOYSA-N 0.000 description 1
- WQZGKKKJIJFFOK-QTVWNMPRSA-N D-mannopyranose Chemical compound OC[C@H]1OC(O)[C@@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-QTVWNMPRSA-N 0.000 description 1
- PNNNRSAQSRJVSB-SLPGGIOYSA-N Fucose Natural products C[C@H](O)[C@@H](O)[C@H](O)[C@H](O)C=O PNNNRSAQSRJVSB-SLPGGIOYSA-N 0.000 description 1
- 241000167880 Hirundinidae Species 0.000 description 1
- 101000616810 Homo sapiens MAL-like protein Proteins 0.000 description 1
- SHZGCJCMOBCMKK-DHVFOXMCSA-N L-fucopyranose Chemical compound C[C@@H]1OC(O)[C@@H](O)[C@H](O)[C@@H]1O SHZGCJCMOBCMKK-DHVFOXMCSA-N 0.000 description 1
- WHXSMMKQMYFTQS-UHFFFAOYSA-N Lithium Chemical compound [Li] WHXSMMKQMYFTQS-UHFFFAOYSA-N 0.000 description 1
- 102100021832 MAL-like protein Human genes 0.000 description 1
- 239000007832 Na2SO4 Substances 0.000 description 1
- 240000002853 Nelumbo nucifera Species 0.000 description 1
- 235000006508 Nelumbo nucifera Nutrition 0.000 description 1
- 235000006510 Nelumbo pentapetala Nutrition 0.000 description 1
- 239000012564 Q sepharose fast flow resin Substances 0.000 description 1
- 235000009754 Vitis X bourquina Nutrition 0.000 description 1
- 235000012333 Vitis X labruscana Nutrition 0.000 description 1
- 240000006365 Vitis vinifera Species 0.000 description 1
- 235000014787 Vitis vinifera Nutrition 0.000 description 1
- 238000005349 anion exchange Methods 0.000 description 1
- PYMYPHUHKUWMLA-UHFFFAOYSA-N arabinose Natural products OCC(O)C(O)C(O)C=O PYMYPHUHKUWMLA-UHFFFAOYSA-N 0.000 description 1
- SRBFZHDQGSBBOR-UHFFFAOYSA-N beta-D-Pyranose-Lyxose Natural products OC1COC(O)C(O)C1O SRBFZHDQGSBBOR-UHFFFAOYSA-N 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 235000014633 carbohydrates Nutrition 0.000 description 1
- 238000001460 carbon-13 nuclear magnetic resonance spectrum Methods 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 229940106157 cellulase Drugs 0.000 description 1
- 239000000470 constituent Substances 0.000 description 1
- 239000000110 cooling liquid Substances 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- CSCPPACGZOOCGX-WFGJKAKNSA-N deuterated acetone Substances [2H]C([2H])([2H])C(=O)C([2H])([2H])[2H] CSCPPACGZOOCGX-WFGJKAKNSA-N 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000004821 distillation Methods 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 230000008014 freezing Effects 0.000 description 1
- 229920000550 glycopolymer Polymers 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 230000002519 immonomodulatory effect Effects 0.000 description 1
- 239000003456 ion exchange resin Substances 0.000 description 1
- 229920003303 ion-exchange polymer Polymers 0.000 description 1
- 238000002356 laser light scattering Methods 0.000 description 1
- 229910052744 lithium Inorganic materials 0.000 description 1
- 238000000874 microwave-assisted extraction Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 229920001542 oligosaccharide Polymers 0.000 description 1
- 150000002482 oligosaccharides Chemical class 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 230000000242 pagocytic effect Effects 0.000 description 1
- 238000002203 pretreatment Methods 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 238000000197 pyrolysis Methods 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000000790 scattering method Methods 0.000 description 1
- 229910052938 sodium sulfate Inorganic materials 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 238000012916 structural analysis Methods 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 150000003538 tetroses Chemical class 0.000 description 1
- 238000000108 ultra-filtration Methods 0.000 description 1
- 238000002525 ultrasonication Methods 0.000 description 1
- 238000001291 vacuum drying Methods 0.000 description 1
Classifications
-
- 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
- C08B37/006—Heteroglycans, i.e. polysaccharides having more than one sugar residue in the main chain in either alternating or less regular sequence; Gellans; Succinoglycans; Arabinogalactans; Tragacanth or gum tragacanth or traganth from Astragalus; Gum Karaya from Sterculia urens; Gum Ghatti from Anogeissus latifolia; Derivatives thereof
- C08B37/0087—Glucomannans or galactomannans; Tara or tara gum, i.e. D-mannose and D-galactose units, e.g. from Cesalpinia spinosa; Tamarind gum, i.e. D-galactose, D-glucose and D-xylose units, e.g. from Tamarindus indica; Gum Arabic, i.e. L-arabinose, L-rhamnose, D-galactose and D-glucuronic acid units, e.g. from Acacia Senegal or Acacia Seyal; Derivatives thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/02—Immunomodulators
-
- 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
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Medicinal Chemistry (AREA)
- Polymers & Plastics (AREA)
- Biochemistry (AREA)
- Materials Engineering (AREA)
- Immunology (AREA)
- Molecular Biology (AREA)
- Pharmacology & Pharmacy (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Emergency Medicine (AREA)
- Animal Behavior & Ethology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- General Chemical & Material Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Sustainable Development (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention belongs to pharmaceutical technology fields, and in particular to a kind of ganoderma lucidum beta glucan and preparation method thereof and prepare the application in immunoregulation medicament.Ganoderma lucidum beta glucan molecular weight is 5~500kDa, monosaccharide group becomes glucose and glucuronic acid, two kinds of monosaccharide ratios are 30:1~4:1, and connection type is with → 3) Glc (β 1 → and → 6) Glc (β 1 → based on, two kinds of connection type ratios are 1:1~3:1.The present invention is by the way that by ganoderma lucidum medicinal material, after superheated water extracts and removes water-soluble polysaccharide, residue is extracted with aqueous slkali again, and after extracting solution is neutralized with hydrochloric acid, concentration obtains Thick many candies after ethanol precipitation.Thick many candies are purified by anion exchange resin, finally obtain the beta glucan of high-purity, can be remarkably promoted RAW264.7 cell by cell experiment proof and be swallowed dimethyl diaminophenazine chloride, can be used for preparing immunoregulatory drug.
Description
Technical field
The invention belongs to pharmaceutical technology fields, and in particular to a kind of ganoderma lucidum beta glucan and preparation method thereof and exempt from preparation
Epidemic disease adjusts the application in drug.
Background technique
Ganoderma lucidum polysaccharide is one of active constituent more important in ganoderma lucidum, and structural analysis shows ganoderma lucidum polysaccharide mainly with high score
Based on the glycopolymers of son amount, mainly by glucose, xylose, mannose, the monosaccharide such as galactolipin and fucose formation ground carbohydrate
Macromolecular composition.Separate sources and extracting mode obtain ground ganoderma lucidum polysaccharide, have different monosaccharide compositions, molecular weight, branch
The features such as conformation and dissolubility can induce panimmunity reaction to play immunomodulatory effect.
Traditional Extraction method of ganoderan is mainly water extract-alcohol precipitation, ultrasonic wave assisted extraction method, microwave―assisted extraction,
Enzymatic Extraction.Chinese patent CN102766221A discloses a kind of extracting method of ganoderma lucidum polysaccharide, and this method includes complex enzyme zymohydrolysis
Ganoderma lucidum fruitbody, ultrasonication and ganoderma lucidum polysaccharide are extracted.Chinese patent CN104292357A discloses a kind of mentioning for ganoderma lucidum polysaccharide
Method is taken, this method includes that polysaccharide is extracted from ganoderma lucidum, will be in ganoderma lucidum using a variety of different enzymes using enzyme effect mild condition
Fat, cellulase hydrolysis, Polyose extraction therein is come out, the utilization rate of ganoderma lucidum is improved.Chinese patent CN104558227A
A kind of extracting method of ganoderma lucidum polysaccharide is disclosed, comprising the following steps: pre-treatment, freezing extraction, alcohol precipitation, removing protein, takes off coarse extraction
Color, ultrafiltration, vacuum drying.Above 3 patents are not related to the research of residue after water proposes.
Ganoderma lucidum is a kind of rare Chinese medicine, however ganoderma lucidum polysaccharide recovery rate is lower is a problem to be solved, the present invention
Residue after mentioning to water is further extracted with lye, improves the bioavailability of ganoderma lucidum.
Summary of the invention
The object of the present invention is to provide a kind of ganoderma lucidum beta glucan and preparation method thereof and preparing immunoregulatory drug
In application.The present invention from red ganoderma water mention after residue in further with the natural beta glucan of the isolated one kind of lye,
And purified with anion-exchange column, the beta glucan of high-purity is finally obtained, proves that this kind of β-Portugal is poly- by cell experiment
Sugar can remarkably promote RAW264.7 cell phagocytosis dimethyl diaminophenazine chloride, can be used for preparing immunoregulatory drug.
For achieving the above object, the present invention adopts the following technical solutions:
A kind of ganoderma lucidum beta glucan, ganoderma lucidum beta glucan molecular weight are 5~500kDa, and monosaccharide group becomes glucose and grape
Uronic acid, two kinds of monosaccharide mass ratios are 30:1~4:1, and connection type is with → 3) Glc (β 1 → and → 6) Glc (β 1 → be
Main, two kinds of connection type ratios are 1:1~3:1.
It is water-soluble more to extract removing by hot water using red ganoderma as raw material for the preparation method of the ganoderma lucidum beta glucan
Sugar, then extracted with lye, after extracting solution is neutralized with hydrochloric acid, concentration obtains Thick many candies through ethanol precipitation；Thick many candies are by yin
Ion-exchange resin purification finally obtains the glucan of high-purity.
The preparation method of the ganoderma lucidum beta glucan, comprising the following steps:
(1) hot water, which extracts, removes water-soluble polysaccharide: red ganoderma be crushed into 60 meshes, be added distilled water, heating extraction 1~
3 times, every time 1~3h, remove water-soluble polysaccharide, after filtering means dehydration, the medicinal material residue of red ganoderma is dried；
(2) it extracts to obtain beta glucan crude product aqueous solution: by medicinal material residue obtained by step (1), aqueous slkali is added and extracts 1~3
It is secondary, after being centrifuged 10~30min with 4000~5000r/min, merge and collect supernatant, the HCl of 0.5~1.5M of molar concentration is added
Aqueous solution is neutralized to supernatant pH value for neutrality, by the 1/2~1/5 of supernatant vacuum-concentrcted to original volume；
(3) ethanol precipitation obtains beta glucan crude extract: into step (2) acquired solution, 2~5 times of volumes are added
The ethyl alcohol of 95wt% purity, is placed at room temperature for 24~72h, with 9000~11000r/min be centrifuged 5~15min separate solid is heavy
It forms sediment, collects precipitating and redissolve freeze-drying to get beta glucan crude extract；
(4) purifying crude obtains refining ganoderam lucidum beta glucan: the distillation of beta glucan crude extract obtained by step (3) is water-soluble
Solution, using sodium-chloride water solution as mobile phase, isolates and purifies by strong anion exchange resin, detects through Phenol-sulphate acid method, merges
The component containing beta glucan is collected, dialyses by distilled water, is concentrated under reduced pressure, is lyophilized, obtain high-purity ganoderma lucidum beta glucan.
The preparation method of the ganoderma lucidum beta glucan, feed liquid mass ratio 1:5~1:20 that hot water extracts in step (1),
Extracting temperature is 60~90 DEG C.
The preparation method of the ganoderma lucidum beta glucan, aqueous slkali is sodium hydroxide, potassium hydroxide, hydrogen-oxygen in step (2)
Change at least one of lithium, sodium carbonate, potassium carbonate, sodium bicarbonate or aqueous solution of ammonium hydroxide, molar concentration are as follows: 0.05~
2M, the feed liquid mass ratio that alkali carries take are as follows: 1:3~1:20, Extracting temperature are as follows: -20 DEG C~60 DEG C, extraction time are as follows: 0.5~4h.
The preparation method of the ganoderma lucidum beta glucan, in step (4) molar concentration of sodium-chloride water solution be 0.1~
2.5M；Anion exchange resin is DEAE-cellulose (DEAE-cellulose), DEAE- glucan (DEAE-
Sephadex), at least one of DEAE- agarose (DEAE-Sepharose) and Q Sephadex Fast Flow.
The ganoderma lucidum beta glucan passes through cell experiment preparing the application in immunoregulation medicament, ganoderma lucidum beta glucan
Proof can remarkably promote RAW264.7 cell phagocytosis dimethyl diaminophenazine chloride, be used to prepare immunoregulatory drug, and be 50~200 in concentration
μ g/ml shows stronger immunoregulatory activity.
Compared with prior art, it advantages of the present invention and has the technical effect that
(1) raw material of the present invention is that red ganoderma water mentions residue, and raw material sources are abundant, securely and reliably, its biology benefit can be improved
With rate.
(2) preparation process that the present invention uses is simple, and Thick many candies yield is up to 1.5~5%.
(3) using the beta glucan of the method for the present invention preparation, product purity is high, and molecular weight is 5~500kDa, monosaccharide composition
For glucose and glucuronic acid, two kinds of monosaccharide ratios are 30:1~4:1, and connection type is with → 3) Glc (β 1 → and → 6)
Glc (β 1 → based on, two kinds of connection type ratios are 1:1~3:1.
(4) RAW264.7 cell can be remarkably promoted by cell experiment proof using beta glucan prepared by the present invention to gulp down
Dimethyl diaminophenazine chloride is bitten, can be used for preparing immunoregulatory drug.
Detailed description of the invention
Fig. 1 is that the GPC of ganoderma lucidum beta glucan of the present invention schemes.In figure, abscissa time represents time (min), ordinate
Relative scale represents relative peak area.
Fig. 2 is the nmr spectrum of ganoderma lucidum beta glucan of the present invention.Wherein, Fig. 2 (a) is the beta glucan1H NMR figure
Spectrum；Fig. 2 (b) is the beta glucan13C NMR spectra；Fig. 2 (c) is the beta glucan1H-1H COSY map；Fig. 2 (d) is the β-
Glucan1H-13C HSQC map.
Fig. 3 for five formed pentasaccharides of glucose of the invention ESI-CID-MS2Figure.In figure, abscissa m/z represents matter lotus
Than ordinate Relative Abundance represents each fragment relative abundance.
Fig. 4 is the influence that ganoderma lucidum beta glucan of the present invention swallows dimethyl diaminophenazine chloride ability to 264.7 cell of mouse RAW.In figure,
Ordinate OD 540nm, which is represented, surveys each hole absorbance value at 540nm.Note: compared with model group, * P < 0.05, * * P < 0.01.
Specific embodiment
Technical solution of the present invention is further described in detail with reference to the accompanying drawings and detailed description, but the present invention
Claimed range is not limited to the range of strength statement.
Embodiment 1: the preparation of ganoderma lucidum beta glucan
The preparation method of ganoderma lucidum beta glucan of the invention specifically includes the following steps:
(1) hot water, which extracts, removes water-soluble polysaccharide: red ganoderma crushed 60 meshes, be added and steam by feed liquid mass ratio 1:10
Distilled water, 80 DEG C heating extraction 2 times, each 2h, remove water-soluble polysaccharide, red ganoderma medicinal material residue drying；
(2) it extracts beta glucan crude product: by medicinal material residue obtained by step (1), adding molar concentration by feed liquid mass ratio 1:10
The NaOH aqueous solution of 2M, 60 DEG C of heating extraction 1h are extracted twice, and centrifugation (4500r/min, 20min), which merges, collects supernatant, are added
The HCL aqueous solution of molar concentration 1M, neutralization supernatant, the 1/3 of vacuum-concentrcted to original volume.
(3) ethanol precipitation: into step (2) acquired solution, the 95wt% pure ethanol of 4 times of volumes is added, is placed at room temperature for
48h is centrifuged (10000r/min, 10min), is collected precipitating and is redissolved freeze-drying to get beta glucan crude extract；
(4) purifying crude: beta glucan crude extract obtained by step (3) is dissolved with distilled water, with molar concentration 0.2M's
Sodium-chloride water solution is mobile phase, is isolated and purified by strong anion exchange resin, Phenol-sulphate acid method detection, merges to collect and contain
Glucan component, dialyses by distilled water, is concentrated under reduced pressure, is lyophilized, and obtains high-purity beta glucan.
Embodiment 2: the structural characterization of ganoderma lucidum beta glucan
The structural characterization of ganoderma lucidum beta glucan of the present invention specifically includes the following steps:
(1) absolute molecular weight measurement and purity analysis: using High Performance Gel Permeation chromatography (HPGPC)-ten octagonal laser light
The absolute molecular quality and purity assay of scattering method (MALLs) combination measurement ganoderma lucidum beta glucan.
Chromatographic condition: chromatographic column: Shodex OHPak SB804HQ chromatographic column and Shodex OHPak SB802.5HQ color
Compose column series connection；Mobile phase: 0.1molL-1Na2SO4Aqueous solution；Composition distribution is connected online with ten octagonal laser light scattering instruments
Detection, measures the molecular weight of beta glucan；Sample is in the single symmetrical peak of GPC spectrogram intermediate range (as shown in Figure 1, the peak after 30min
For NaCl), sample purity is very high, and measuring its absolute molecular quality is 24.7kDa.
(2) nuclear magnetic resoance spectrum map analysis: being transferred in nuclear magnetic tube after taking this kind of beta glucan of 20mg to carry out heavy water exchange, with
Deuterated acetone is internal standard, in 25 DEG C of progress nmr analysis.As a result such as Fig. 2 (a)-(d), this kind of beta glucan is mainly β-
The glucan of (1 → 3) and β-(1 → 6) connection, residue segment mainly include three kinds of forms, A: β-(1 → 6)-Glcp, and B: β-(1
→ 3)-Glcp, C: β-T-Glcp.The chemical displacement value of three kinds of glucose residues is concluded as shown in table 1.
The chemical displacement value of 1 three kinds of saccharide residue segments of table concludes table
(3)ESI-CID-MS2Analysis: by this kind of beta glucan with 5~20 times of amounts (W/V, bulking values under heating stirring
Than) concentration 0.01mol/L~2.0mol/L diluted acid 6~8h of degradation, alkali neutralization, concentration, supernatant after centrifugation are used after reaction solution is cooling
Liquid is concentrated under reduced pressure to give sour water solution oligosaccharide mixture.The mixture passes through second mass analysis, and wherein oligose fragment is relatively more
Be pentasaccharides, tetrose and six sugar, these three oligose fragments ratio column be respectively 16.8%, 13.8% and 12.4%.Pentasaccharides
ESI-CID-MS2Map is as shown in figure 3, be glucose A at mass-to-charge ratio (m/z) 768.280,2It is broken segment peak, it was demonstrated that glucose 2
Position is glucose B at 708.11 without being substituted0,2It is broken segment peak, the glycosidic bond fracture generation between glucose at 665.10
C segment peak, be glucose B at 588.320,2, D0,2The segment peak of generation is broken at two simultaneously, is glucose E at 384.380,2
Be broken segment peak, at 341.34 for glucose between glycosidic bond fracture generation F segment peak.Be computed molecular weight (Mw)=
828.27, be five glucose groups at pentasaccharides survey molecular weight, it is consistent with its 828.8 dalton of theoretical molecular weight, be by 5
Glc composition, structure sequence are Glc- (Glc)2-(Glc)2。
Embodiment 3: the immunoregulatory activity measurement of ganoderma lucidum beta glucan
The immunoregulatory activity measurement of ganoderma lucidum beta glucan of the present invention specifically includes following procedure:
RAW264.7 cell is cultivated, 96 orifice plates are planted, every hole cell number is 20,000, and dosing culture is for 24 hours.After for 24 hours, supernatant is abandoned,
PBS is washed cell 3 times, and the 100 μ L that concentration is 0.075wt% are added in every hole, after being incubated for 20min, abandons supernatant, and PBS is washed cell 3 times,
Every hole, which is added at cell pyrolysis liquid 200 μ L, 540nm, surveys each hole absorbance value.Meanwhile mtt assay survey tested material is thin to RAW264.7
The effect of born of the same parents' proliferation, phagocytic activity survey cell proliferation experiment with RAW264.7 cell phagocytosis dimethyl diaminophenazine chloride experiment absorbance and mtt assay
Absorbance ratio indicate.
Influence of the ganoderma lucidum beta glucan to 264.7 cell of mouse RAW phagocytosis dimethyl diaminophenazine chloride ability with LPS as shown in figure 4, made
For positive control, the results showed that, LPS positive controls phagocytosis dimethyl diaminophenazine chloride ratio is 117% (P < 0.05) compared with model group,
264.7 cell of RAW phagocytosis dimethyl diaminophenazine chloride ratio is respectively 117%, 111% and 112% after LZJ-0.2 is handled；.
To sum up, the present invention by by ganoderma lucidum medicinal material after superheated water extracts and removes water-soluble polysaccharide, then with 2M sodium hydroxide
It is extracted at 60 DEG C, after extracting solution is neutralized with hydrochloric acid, concentration obtains Thick many candies through ethanol precipitation.Thick many candies pass through Q-
Sepharose Fast Flow column purification, finally obtains the ganoderma lucidum beta glucan of high-purity, and ganoderma lucidum β-Portugal that the present invention obtains is poly-
Glycan molecule amount is 5~500kDa, and monosaccharide group becomes glucose and glucuronic acid, and two kinds of monosaccharide ratios are 30:1~4:1, is connected
Connecing mode is → 3) Glc (β 1 → and → 6) Glc (β 1 →, two kinds of connection type ratios are 1:1~3:1.Prepared by the present invention
Ganoderma lucidum beta glucan can remarkably promote RAW264.7 cell by cell experiment proof and swallow dimethyl diaminophenazine chloride, can be used for preparing immune tune
The drug of section.
The above examples are only used to illustrate the technical scheme of the present invention, rather than is limited；Although referring to aforementioned reality
Applying example, the present invention will be described in detail, for those of ordinary skill in the art, still can be to previous embodiment
Documented technical solution is modified or equivalent replacement of some of the technical features；And these modifications or substitutions,
The spirit and scope for claimed technical solution of the invention that it does not separate the essence of the corresponding technical solution.
Claims (8)
1. a kind of ganoderma lucidum beta glucan, which is characterized in that ganoderma lucidum beta glucan molecular weight is 5~500kDa, and monosaccharide group becomes Portugal
Grape sugar and glucuronic acid, two kinds of monosaccharide mass ratios are 30:1~4:1, and connection type is with → 3) Glc (β 1 → and → 6)
Glc (β 1 → based on, two kinds of connection type ratios are 1:1~3:1.
2. a kind of preparation method of ganoderma lucidum beta glucan described in claim 1, which is characterized in that using red ganoderma as raw material, lead to
Superheated water, which extracts, removes water-soluble polysaccharide, then is extracted with lye, after extracting solution is neutralized with hydrochloric acid, concentration, through ethanol precipitation
Obtain Thick many candies；Thick many candies are purified by anion exchange resin, finally obtain the glucan of high-purity.
3. the preparation method of ganoderma lucidum beta glucan according to claim 2, which comprises the following steps:
(1) hot water, which extracts, removes water-soluble polysaccharide: red ganoderma be crushed into 60 meshes, be added distilled water, heating extraction 1~3 time,
1~3h every time removes water-soluble polysaccharide, and after filtering means dehydration, the medicinal material residue of red ganoderma is dried；
(2) it extracts to obtain beta glucan crude product aqueous solution: by medicinal material residue obtained by step (1), aqueous slkali is added and extracts 1~3 time, with
After 4000~5000r/min is centrifuged 10~30min, merges and collect supernatant, the HCl/water that 0.5~1.5M of molar concentration is added is molten
Liquid is neutralized to supernatant pH value for neutrality, by the 1/2~1/5 of supernatant vacuum-concentrcted to original volume；
(3) ethanol precipitation obtains beta glucan crude extract: into step (2) acquired solution, the 95wt% that 2~5 times of volumes are added is pure
The ethyl alcohol of degree, is placed at room temperature for 24~72h, is centrifuged 5~15min with 9000~11000r/min and separates to obtain solid precipitating, it is heavy to collect
It forms sediment and redissolves freeze-drying to get beta glucan crude extract；
(4) purifying crude obtains refining ganoderam lucidum beta glucan: beta glucan crude extract obtained by step (3) is dissolved with distilled water, with
Sodium-chloride water solution is mobile phase, is isolated and purified by strong anion exchange resin, is detected through Phenol-sulphate acid method, merges collection and contains
There is the component of beta glucan, dialyse by distilled water, be concentrated under reduced pressure, be lyophilized, obtains high-purity ganoderma lucidum beta glucan.
4. the preparation method of ganoderma lucidum beta glucan according to claim 3, which is characterized in that hot water extracts in step (1)
Feed liquid mass ratio 1:5~1:20, Extracting temperature be 60~90 DEG C.
5. the preparation method of ganoderma lucidum beta glucan according to claim 3, which is characterized in that aqueous slkali is in step (2)
Sodium hydroxide, potassium hydroxide, lithium hydroxide, sodium carbonate, potassium carbonate, sodium bicarbonate or ammonium hydroxide aqueous solution at least one
Kind, molar concentration are as follows: 0.05~2M, the feed liquid mass ratio that alkali carries take are as follows: 1:3~1:20, Extracting temperature are as follows: -20 DEG C~60
DEG C, extraction time are as follows: 0.5~4h.
6. the purification process of ganoderma lucidum beta glucan according to claim 3, which is characterized in that sodium chloride water in step (4)
The molar concentration of solution is 0.1~2.5M；Anion exchange resin is DEAE-cellulose (DEAE-
Cellulose), DEAE- glucan (DEAE-Sephadex), DEAE- agarose (DEAE-Sepharose) and Q Sephadex
At least one of Fast Flow.
7. ganoderma lucidum beta glucan described in a kind of one of claims 1 to 6 is preparing the application in immunoregulation medicament.
8. ganoderma lucidum beta glucan according to claim 7 is preparing the application in immunoregulatory drug, which is characterized in that
Ganoderma lucidum beta glucan can remarkably promote RAW264.7 cell by cell experiment proof and swallow dimethyl diaminophenazine chloride, be used to prepare immunological regulation
Drug, and concentration be 50~200 μ g/ml show stronger immunoregulatory activity.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810426702.4A CN108976314A (en) | 2018-05-07 | 2018-05-07 | A kind of ganoderma lucidum beta glucan and preparation method thereof and preparing the application in immunoregulation medicament |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810426702.4A CN108976314A (en) | 2018-05-07 | 2018-05-07 | A kind of ganoderma lucidum beta glucan and preparation method thereof and preparing the application in immunoregulation medicament |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN108976314A true CN108976314A (en) | 2018-12-11 |
Family
ID=64542507
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810426702.4A Pending CN108976314A (en) | 2018-05-07 | 2018-05-07 | A kind of ganoderma lucidum beta glucan and preparation method thereof and preparing the application in immunoregulation medicament |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108976314A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112979840A (en) * | 2021-04-25 | 2021-06-18 | 上海市农业科学院 | Method for separating and purifying ganoderma lucidum beta-glucooligosaccharides |
| CN115558037A (en) * | 2022-10-19 | 2023-01-03 | 陕西科技大学 | Ganoderma lucidum beta-glucan extract and preparation method and detection method thereof |
| CN117186259A (en) * | 2023-08-04 | 2023-12-08 | 深圳市岩代投资有限公司 | Polysaccharide compound with definite molecular structure and capable of eliminating toxic and side effects of chemotherapeutic drugs |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1454904A (en) * | 2003-05-08 | 2003-11-12 | 扬州大学 | Method of preparing high purity fungus polysaccharide |
| TWI281399B (en) * | 2001-08-06 | 2007-05-21 | Academia Sinica | Immuno-modulating antitumor activities of ganoderma lucidum (Reishi) polysaccharides |
| CN101805414A (en) * | 2010-04-06 | 2010-08-18 | 无限极（中国）有限公司 | Preparation method of ganoderma lucidum polysaccharide with high yield |
| CN106832042A (en) * | 2017-02-03 | 2017-06-13 | 中国科学院上海药物研究所 | The glucans of β 1,3, its preparation method and pharmaceutical applications |
-
2018
- 2018-05-07 CN CN201810426702.4A patent/CN108976314A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| TWI281399B (en) * | 2001-08-06 | 2007-05-21 | Academia Sinica | Immuno-modulating antitumor activities of ganoderma lucidum (Reishi) polysaccharides |
| CN1454904A (en) * | 2003-05-08 | 2003-11-12 | 扬州大学 | Method of preparing high purity fungus polysaccharide |
| CN101805414A (en) * | 2010-04-06 | 2010-08-18 | 无限极（中国）有限公司 | Preparation method of ganoderma lucidum polysaccharide with high yield |
| CN106832042A (en) * | 2017-02-03 | 2017-06-13 | 中国科学院上海药物研究所 | The glucans of β 1,3, its preparation method and pharmaceutical applications |
Non-Patent Citations (7)
| Title |
|---|
| 刘程等主编: "《当代新型食品》", 31 December 1998, 北京工业大学出版社 * |
| 周荣汉等主编: "《植物化学分类》", 31 March 2013, 中国中医药出版社 * |
| 张俐娜等: ""灵芝子实体水溶性多糖的分离和分子量测定"", 《高分子学报》 * |
| 段金廒等主编: "《中药资源化学》", 30 September 2013, 中国中医药出版社 * |
| 潘道东主编: "《功能性食品添加剂》", 31 January 2006, 中国轻工业出版社 * |
| 王琪: ""灵芝子实体多糖的分离纯化、硫酸化及生物活性研究"", 《中国优秀硕士学位论文全文数据库 农业科技辑》 * |
| 陈康林著: "《野生灵芝 国药之王》", 31 July 2005, 中国科学技术出版社 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112979840A (en) * | 2021-04-25 | 2021-06-18 | 上海市农业科学院 | Method for separating and purifying ganoderma lucidum beta-glucooligosaccharides |
| CN112979840B (en) * | 2021-04-25 | 2022-07-29 | 上海市农业科学院 | Method for separating and purifying ganoderma lucidum beta-glucooligosaccharides |
| CN115558037A (en) * | 2022-10-19 | 2023-01-03 | 陕西科技大学 | Ganoderma lucidum beta-glucan extract and preparation method and detection method thereof |
| CN115558037B (en) * | 2022-10-19 | 2024-02-06 | 陕西科技大学 | Ganoderma lucidum beta-glucan extract and preparation method and detection method thereof |
| CN117186259A (en) * | 2023-08-04 | 2023-12-08 | 深圳市岩代投资有限公司 | Polysaccharide compound with definite molecular structure and capable of eliminating toxic and side effects of chemotherapeutic drugs |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN109400734B (en) | Rosa roxburghii polysaccharide and preparation method and application thereof | |
| CN102336840B (en) | Triple-helical Tremellan, preparation method and application thereof | |
| CN105001352B (en) | Beta-1,3/1,6-glucan, preparation method therefor, and application thereof in preparing immune enhancement and anti-tumor medicine and functional food | |
| CN108976314A (en) | A kind of ganoderma lucidum beta glucan and preparation method thereof and preparing the application in immunoregulation medicament | |
| Yang et al. | Isolation, purification, structural characterization, and hypoglycemic activity assessment of polysaccharides from Hovenia dulcis (Guai Zao) | |
| Li et al. | Isolation and structural characterization of a neutral polysaccharide from the stems of Dendrobium densiflorum | |
| Liu et al. | Structure and chain conformation of bioactive β-D-glucan purified from water extracts of Ganoderma lucidum unbroken spores | |
| CN109912700A (en) | A method of utilizing β -1,6 dextranase coproduction yeast dextran, Mannoproteins and yeast extract | |
| CN114591448B (en) | Mulberry and phellinus linteus fruiting body manna galactosan and preparation and application thereof | |
| Shi et al. | Structural characterization of a sulfated glucan isolated from the aqueous extract of Hedysarum polybotrys Hand.-Mazz | |
| CN111892663B (en) | Hericium erinaceus polysaccharide and preparation method and application thereof | |
| CN111978421A (en) | Phellinus igniarius polysaccharide and preparation and application thereof | |
| Yang et al. | Separation, characterization and hypoglycemic activity in vitro evaluation of a low molecular weight heteropolysaccharide from the fruiting body of Phellinus pini | |
| CN110916198A (en) | Method for simultaneously preparing pectic polysaccharide and viscous glycoprotein by using okra fermented wine lees | |
| Wang et al. | Purification, structure identification and immune activity of a neutral polysaccharide from Cynanchum auriculatum | |
| CN113603732B (en) | Non-animal chondroitin sulfate oligosaccharide and preparation method thereof | |
| Zong et al. | Optimized acid hydrolysis conditions for better characterization the structure of inulin-type fructan from Polygonatum sibiricum | |
| CN110128489A (en) | A method of galactomannan oligosaccharide is prepared from hydrolysis | |
| CN108892732B (en) | Preparation method and application of Jinchang jujube polysaccharide with immunoregulation function | |
| CN116874622A (en) | Armillariella mellea polysaccharide and preparation method and application thereof | |
| CN114316080B (en) | Method for improving extraction rate and bioactivity of grifola frondosa crude polysaccharide | |
| CN108976313A (en) | A kind of polyglucose aldehydic acid and preparation method thereof and the application in immunoregulation medicament | |
| CN115477709A (en) | Water-soluble yeast beta-glucan and preparation method thereof | |
| CN110343156B (en) | Extraction and purification method of black bean glycoprotein | |
| YANG et al. | Community structure and plant diversity under different degrees of degraded grassland in Tianzhu, Gansu |
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 | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20181211 |
|
| RJ01 | Rejection of invention patent application after publication |