CN113151372A - Preparation process of high-purity ganoderma lucidum polysaccharide - Google Patents
Preparation process of high-purity ganoderma lucidum polysaccharide Download PDFInfo
- Publication number
- CN113151372A CN113151372A CN202110538939.3A CN202110538939A CN113151372A CN 113151372 A CN113151372 A CN 113151372A CN 202110538939 A CN202110538939 A CN 202110538939A CN 113151372 A CN113151372 A CN 113151372A
- Authority
- CN
- China
- Prior art keywords
- ganoderma lucidum
- mycelia
- centrifuging
- purity
- purified water
- 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.)
- Granted
Links
- 240000008397 Ganoderma lucidum Species 0.000 title claims abstract description 73
- 235000001637 Ganoderma lucidum Nutrition 0.000 title claims abstract description 73
- 150000004676 glycans Chemical class 0.000 title claims abstract description 27
- 229920001282 polysaccharide Polymers 0.000 title claims abstract description 27
- 239000005017 polysaccharide Substances 0.000 title claims abstract description 27
- 238000002360 preparation method Methods 0.000 title claims abstract description 14
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 46
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 37
- 239000008213 purified water Substances 0.000 claims abstract description 34
- 238000000034 method Methods 0.000 claims abstract description 17
- 238000004519 manufacturing process Methods 0.000 claims abstract description 14
- 239000012535 impurity Substances 0.000 claims abstract description 13
- 230000008569 process Effects 0.000 claims abstract description 7
- 238000005119 centrifugation Methods 0.000 claims abstract description 3
- 238000000855 fermentation Methods 0.000 claims description 29
- 230000004151 fermentation Effects 0.000 claims description 29
- 239000006228 supernatant Substances 0.000 claims description 24
- 238000009835 boiling Methods 0.000 claims description 19
- 241000222336 Ganoderma Species 0.000 claims description 13
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 12
- 239000000843 powder Substances 0.000 claims description 12
- 239000002244 precipitate Substances 0.000 claims description 12
- 230000001376 precipitating effect Effects 0.000 claims description 12
- 238000003756 stirring Methods 0.000 claims description 12
- 239000002609 medium Substances 0.000 claims description 10
- 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 description 8
- 239000008103 glucose Substances 0.000 claims description 8
- 239000007788 liquid Substances 0.000 claims description 7
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims description 6
- 239000002518 antifoaming agent Substances 0.000 claims description 6
- 238000012258 culturing Methods 0.000 claims description 6
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 6
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 6
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 6
- 235000019796 monopotassium phosphate Nutrition 0.000 claims description 6
- 238000000643 oven drying Methods 0.000 claims description 6
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 claims description 6
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 claims description 6
- 239000001963 growth medium Substances 0.000 claims description 2
- 238000000605 extraction Methods 0.000 abstract description 12
- 230000008901 benefit Effects 0.000 abstract description 6
- 239000003814 drug Substances 0.000 abstract description 5
- 238000002474 experimental method Methods 0.000 abstract description 4
- 239000003960 organic solvent Substances 0.000 abstract description 4
- 102000004190 Enzymes Human genes 0.000 abstract description 3
- 108090000790 Enzymes Proteins 0.000 abstract description 3
- 238000003809 water extraction Methods 0.000 abstract description 2
- 210000002421 cell wall Anatomy 0.000 abstract 1
- 238000001035 drying Methods 0.000 abstract 1
- 230000003834 intracellular effect Effects 0.000 abstract 1
- 238000001556 precipitation Methods 0.000 abstract 1
- 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 description 8
- 229960001031 glucose Drugs 0.000 description 8
- 206010028980 Neoplasm Diseases 0.000 description 4
- 238000002835 absorbance Methods 0.000 description 4
- OQUKIQWCVTZJAF-UHFFFAOYSA-N phenol;sulfuric acid Chemical compound OS(O)(=O)=O.OC1=CC=CC=C1 OQUKIQWCVTZJAF-UHFFFAOYSA-N 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 241000233866 Fungi Species 0.000 description 2
- 230000009471 action Effects 0.000 description 2
- 230000032683 aging Effects 0.000 description 2
- 230000000259 anti-tumor effect Effects 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 230000036039 immunity Effects 0.000 description 2
- 230000002401 inhibitory effect Effects 0.000 description 2
- 230000001105 regulatory effect Effects 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 238000009098 adjuvant therapy Methods 0.000 description 1
- 230000002429 anti-coagulating effect Effects 0.000 description 1
- 230000003110 anti-inflammatory effect Effects 0.000 description 1
- 230000002785 anti-thrombosis Effects 0.000 description 1
- 230000000840 anti-viral effect Effects 0.000 description 1
- 239000003146 anticoagulant agent Substances 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- -1 antitumor Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 230000005907 cancer growth Effects 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 230000002124 endocrine Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 238000006911 enzymatic reaction Methods 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 210000002865 immune cell Anatomy 0.000 description 1
- 230000036737 immune function Effects 0.000 description 1
- 210000000987 immune system Anatomy 0.000 description 1
- 230000007365 immunoregulation Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 231100000053 low toxicity Toxicity 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 239000010413 mother solution Substances 0.000 description 1
- 210000005036 nerve Anatomy 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 238000000638 solvent extraction Methods 0.000 description 1
- 239000012086 standard solution Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-N sulfuric acid Substances OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
Images
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
-
- 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
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Biochemistry (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Molecular Biology (AREA)
- General Chemical & Material Sciences (AREA)
- General Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Genetics & Genomics (AREA)
- Sustainable Development (AREA)
- Microbiology (AREA)
- Materials Engineering (AREA)
- Medicinal Chemistry (AREA)
- Polymers & Plastics (AREA)
- Medicines Containing Plant Substances (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
Abstract
The invention discloses a preparation process of high-purity ganoderma lucidum polysaccharide, which comprises the steps of fermenting ganoderma lucidum mycelia, removing extracellular impurities, cell wall impurities and part of intracellular impurities, adding isometric purified water into the ganoderma lucidum mycelia after the impurities are removed, placing the ganoderma lucidum mycelia into a wall breaking machine for mechanical crushing, hot water extraction, alcohol precipitation, redissolution and centrifugation, and drying to obtain the high-purity ganoderma lucidum polysaccharide. The process method provided by the invention does not use enzymes, organic solvents and the like, does not need high-pressure and high-temperature equipment, and has the advantages of simple extraction conditions, easy implementation, low production cost and strong operability; the extraction purity is up to more than 99 percent, and the obtained pure product of the ganoderma lucidum polysaccharide can be directly applied to various ways such as experiments, medicine, production and the like.
Description
Technical Field
The invention relates to the technical field of bioengineering, in particular to a preparation process of high-purity ganoderma lucidum polysaccharide.
Background
Ganoderma lucidum (Ganoderma spp.) is one of the best-known medicinal fungi in China, has wide pharmacological action and extremely low toxicity, can be clinically used for adjuvant therapy of various diseases, and has the functions of inhibiting tumor, enhancing immunity, delaying aging, protecting liver and the like. The research shows that the compositions of the fruiting body and the mycelium of the ganoderma lucidum have no obvious difference. Traditionally, Ganoderma lucidum is obtained by field collection or artificial cultivation. However, the formation period of the ganoderma lucidum fruiting body is long, and the required labor intensity is high. With the development of liquid submerged fermentation technology, many large-scale fungi have been cultured on a large scale.
Ganoderan is the main active ingredient in ganoderma lucidum mycelia, and has the functions of resisting tumor, regulating immunity, reducing blood fat, reducing blood sugar, resisting oxidation, resisting aging and the like. The ganoderan can achieve the purpose of resisting tumor by regulating the immune function of the organism. The immunoregulation function of ganoderan is the basis of various functions, and the realization of the immune enhancement function and immune recovery function mainly depends on two aspects: one is the result of direct action on immune cells; the other is realized through the mutual regulation of the nerve, the endocrine and the immune system of the body.
The ganoderan has antiviral, antiinflammatory, antitumor, antioxidant, antitumor, antithrombotic and anticoagulant effects. In recent years, the ganoderma lucidum polysaccharide has the effect of inhibiting the growth of cancer cells, and provides a new idea for treating cancer.
The existing hot water extraction method is a more conventional ganoderma lucidum polysaccharide extraction method, but the purity of the ganoderma lucidum polysaccharide obtained by the method is not high, and is about 75%; the ganoderma lucidum polysaccharide with the purity can not be directly applied to medicines, so that the application of the ganoderma lucidum polysaccharide is limited; the enzymatic method and the organic solvent extraction method have certain requirements on extraction conditions and extraction equipment, the production cost is high, and the obtained purity is not ideal.
Therefore, the extraction research of the high-purity harmless ganoderma lucidum polysaccharide becomes important social significance.
Disclosure of Invention
Aiming at the defects of the prior art, the invention aims to provide a preparation process of high-purity ganoderma lucidum polysaccharide, and the process method does not use enzyme, organic solvent and the like, does not need high-pressure and high-temperature equipment, and has the advantages of simple extraction condition, easy implementation, low production cost and strong operability; the extraction purity is up to more than 99 percent, and the obtained pure product of the ganoderma lucidum polysaccharide can be directly applied to various ways such as experiments, medicine, production and the like.
In order to achieve the purpose, the invention provides the following technical scheme:
a preparation process of high-purity ganoderma lucidum polysaccharide comprises the following steps:
(1) firstly preparing a fermentation culture medium, and then culturing in a fermentation tank under the culture conditions of: inoculating Ganoderma mycelia 10%, fermenting at 29 deg.C, stirring at 150rpm/min, fermenting at pH5.7 under 0.05Mpa, and fermenting to obtain Ganoderma fermentation liquid;
(2) centrifuging the ganoderma lucidum fermentation liquor obtained in the step (2) to collect mycelia, adding purified water, uniformly stirring, centrifuging to collect ganoderma lucidum mycelia, and repeating the step for three times;
(3) adding purified water into the ganoderma lucidum mycelia collected in the step (2), boiling for 2h, centrifuging to collect ganoderma lucidum mycelia, and repeating for three times to obtain clear and transparent supernatant;
(4) adding purified water into the ganoderma lucidum mycelia after the impurities are removed in the step (3), mechanically crushing the ganoderma lucidum mycelia in a wall breaking machine for 2min, adding four times of volume of purified water, boiling for 2h, centrifuging, removing the precipitate, and taking supernatant; adding ethanol with the same volume, precipitating with ethanol for 8h, centrifuging to remove precipitate, boiling with purified water for dissolving, centrifuging, and collecting supernatant; precipitating with ethanol, re-dissolving, centrifuging, collecting supernatant for three times to obtain pure ganoderan, and oven drying or lyophilizing to obtain high-purity ganoderan dry powder.
Further, the fermentation medium in the step (1) comprises the following components: 3 percent of glucose, 1 percent of yeast powder, 0.4 percent of monopotassium phosphate, 0.4 percent of magnesium sulfate, 0.2 thousandth of defoaming agent and the balance of water.
Further, the fermentation period of the step (1) is 4-5 days.
Further, the rotation speed of the centrifugation in the step (2) is 4000 r/min.
Further, the volume ratio of the purified water to the ganoderma lucidum mycelia in the step (3) is 5: 1.
further, step (3) boils for 2 h.
Further, the rotating speed of the wall breaking machine in the step (4) during mechanical crushing is 25000 r/min.
The invention has the beneficial effects that:
(1) according to the preparation process provided by the invention, impurities are removed from the ganoderma lucidum mycelia, and then the impurities are simultaneously removed in the extraction process of the ganoderma lucidum polysaccharide through the improvement of the method, so that other impurities are fundamentally prevented from being introduced in the extraction process, and the purity of the product is ensured;
(2) the preparation method provided by the invention does not use enzymes, organic solvents and the like, does not need high-pressure and high-temperature equipment, and has the advantages of simple extraction conditions, easy implementation, low production cost and strong operability.
(3) By adopting the preparation process provided by the invention, the purity of the extracted ganoderma lucidum polysaccharide reaches more than 99 percent, a leap in the field is realized, and the obtained pure ganoderma lucidum polysaccharide can be directly used in various ways such as experiments, medicine, production and the like.
(4) The preparation method provided by the invention does not introduce any harmful substance in the extraction process, has high product purity and greatly reduced cost, and has better economic and social benefits.
Description of the drawings:
FIG. 1 is a process flow diagram of the present invention,
FIG. 2 is a purity standard curve according to the present invention.
Detailed Description
The present invention is further described in detail with reference to the following examples, which are provided for those skilled in the art to fully understand the present invention and are not intended to limit the present invention.
The invention is further illustrated by the following examples:
example 1 a process for preparing high purity ganoderan comprises the steps of:
(1) firstly, preparing a fermentation medium, wherein the medium comprises the following components: 3% of glucose, 1% of yeast powder, 0.4% of monopotassium phosphate, 0.4% of magnesium sulfate, 0.2% of defoaming agent and the balance of water; then culturing in a fermentation tank under the following conditions: inoculating Ganoderma mycelia 10%, fermenting at 29 deg.C, stirring at 150rpm/min, fermenting at pH5.7 under 0.05Mpa for 5 days, and fermenting to obtain Ganoderma fermentation liquid;
(2) centrifuging the ganoderma lucidum fermentation liquor obtained in the step (2) at 4000r/min to collect mycelia, adding purified water, uniformly stirring, centrifuging at 4000r/min to collect ganoderma lucidum mycelia, and repeating the step for three times;
(3) adding purified water with five times of volume into the ganoderma lucidum mycelia collected in the step (2), boiling for 2h, centrifuging and collecting the ganoderma lucidum mycelia, repeating for three times to obtain clear and transparent supernatant;
(4) adding purified water into the ganoderma lucidum mycelia after impurity removal in the step (3), mechanically crushing the ganoderma lucidum mycelia in a wall breaking machine at 25000 r/min for 2min, adding four times of volume of purified water, boiling for 2h, centrifuging, removing precipitate, and taking supernatant; adding ethanol with the same volume, precipitating with ethanol for 8h, centrifuging to remove precipitate, boiling with purified water for dissolving, centrifuging, and collecting supernatant; precipitating with ethanol, re-dissolving, centrifuging, collecting supernatant for three times to obtain pure ganoderan, and oven drying or lyophilizing to obtain high-purity ganoderan dry powder.
Embodiment 2 a process for preparing high purity ganoderan, comprising the steps of:
(1) firstly, preparing a fermentation medium, wherein the medium comprises the following components: 3% of glucose, 1% of yeast powder, 0.4% of monopotassium phosphate, 0.4% of magnesium sulfate, 0.2% of defoaming agent and the balance of water; then culturing in a fermentation tank under the following conditions: inoculating Ganoderma mycelia 10%, fermenting at 29 deg.C, stirring at 150rpm/min, fermenting at pH5.7 under 0.05Mpa for 5 days, and fermenting to obtain Ganoderma fermentation liquid;
(2) centrifuging the ganoderma lucidum fermentation liquor obtained in the step (2) at 4000r/min to collect mycelia, adding purified water, uniformly stirring, centrifuging at 4000r/min to collect ganoderma lucidum mycelia, and repeating the step for three times;
(3) adding purified water with five times of volume into the ganoderma lucidum mycelia collected in the step (2), boiling for 2.5h, centrifuging and collecting the ganoderma lucidum mycelia, repeating for three times to obtain clear and transparent supernatant;
(4) adding purified water into the ganoderma lucidum mycelia after impurity removal in the step (3), mechanically crushing the ganoderma lucidum mycelia in a wall breaking machine at 25000 r/min for 2min, adding four times of volume of purified water, boiling for 2.5h, centrifuging, removing precipitate, and taking supernatant; adding ethanol with the same volume, precipitating with ethanol for 8h, centrifuging to remove precipitate, boiling with purified water for dissolving, centrifuging, and collecting supernatant; precipitating with ethanol, re-dissolving, centrifuging, collecting supernatant for three times to obtain pure ganoderan, and oven drying or lyophilizing to obtain high-purity ganoderan dry powder.
Embodiment 3 a process for preparing high purity ganoderan, comprising the steps of:
(1) firstly, preparing a fermentation medium, wherein the medium comprises the following components: 3% of glucose, 1% of yeast powder, 0.4% of monopotassium phosphate, 0.4% of magnesium sulfate, 0.2% of defoaming agent and the balance of water; then culturing in a fermentation tank under the following conditions: inoculating Ganoderma mycelia 10%, fermenting at 29 deg.C, stirring at 150rpm/min, fermenting at pH5.7 under 0.05Mpa for 4 days, and fermenting to obtain Ganoderma fermentation liquid;
(2) centrifuging the ganoderma lucidum fermentation liquor obtained in the step (2) at 4000r/min to collect mycelia, adding purified water, uniformly stirring, centrifuging at 4000r/min to collect ganoderma lucidum mycelia, and repeating the step for three times;
(3) adding purified water with five times of volume into the ganoderma lucidum mycelia collected in the step (2), boiling for 2.5h, centrifuging and collecting the ganoderma lucidum mycelia, repeating for three times to obtain clear and transparent supernatant;
(4) adding purified water into the ganoderma lucidum mycelia after impurity removal in the step (3), mechanically crushing the ganoderma lucidum mycelia in a wall breaking machine at 25000 r/min for 2min, adding four times of volume of purified water, boiling for 2.5h, centrifuging, removing precipitate, and taking supernatant; adding ethanol with the same volume, precipitating with ethanol for 8h, centrifuging to remove precipitate, boiling with purified water for dissolving, centrifuging, and collecting supernatant; precipitating with ethanol, re-dissolving, centrifuging, collecting supernatant for three times to obtain pure ganoderan, and oven drying or lyophilizing to obtain high-purity ganoderan dry powder.
Example 4 a process for preparing high purity ganoderan comprises the following steps:
(1) firstly, preparing a fermentation medium, wherein the medium comprises the following components: 3% of glucose, 1% of yeast powder, 0.4% of monopotassium phosphate, 0.4% of magnesium sulfate, 0.2% of defoaming agent and the balance of water; then culturing in a fermentation tank under the following conditions: inoculating Ganoderma mycelia 10%, fermenting at 29 deg.C, stirring at 150rpm/min, fermenting at pH5.7 under 0.05Mpa for 5 days, and fermenting to obtain Ganoderma fermentation liquid;
(2) centrifuging the ganoderma lucidum fermentation liquor obtained in the step (2) at 4000r/min to collect mycelia, adding purified water, uniformly stirring, centrifuging at 4000r/min to collect ganoderma lucidum mycelia, and repeating the step for three times;
(3) adding purified water with five times of volume into the ganoderma lucidum mycelia collected in the step (2), boiling for 2h, centrifuging and collecting the ganoderma lucidum mycelia, repeating for three times to obtain clear and transparent supernatant;
(4) adding purified water into the ganoderma lucidum mycelia after impurity removal in the step (3), mechanically crushing the ganoderma lucidum mycelia in a wall breaking machine at 25000 r/min for 2min, adding four times of volume of purified water, boiling for 2h, centrifuging, removing precipitate, and taking supernatant; adding ethanol with the same volume, precipitating with ethanol for 8.5h, centrifuging to remove precipitate, boiling with purified water for dissolving, centrifuging, and collecting supernatant; precipitating with ethanol, re-dissolving, centrifuging, collecting supernatant for three times to obtain pure ganoderan, and oven drying or lyophilizing to obtain high-purity ganoderan dry powder.
Experimental example a purity verification test of ganoderan obtained by the preparation method of the present invention:
the total sugar content in the sample is measured by a sulfuric acid-phenol method to show the purity of the ganoderma lucidum polysaccharide.
(1) Standard curve was determined with glucose as standard:
0.52204g of anhydrous glucose (dried in an oven for 4 hours before weighing) were precisely weighed, dissolved in 25ML water, and diluted 20-fold to obtain 1044. mu.g/mL of a mother solution, and glucose standard solutions of 1+9, 1+11.5, 0.5+9.5, 1+49, 1+ 99-fold, 10-fold, 12.5-fold, 20-fold, 50-fold and 100-fold were obtained, respectively, 104.40. mu.g/mL, 83.52. mu.g/mL, 52.20. mu.g/mL, 20.88. mu.g/mL and 10.44. mu.g/mL were obtained, and the total sugar content was determined by the sulfuric acid-phenol method, respectively:
firstly, sucking 2ml of sample to be detected (taking 2ml of pure water for blank); adding 5ml of concentrated sulfuric acid, and shaking up; ③ adding 2ml of 6 percent phenol, and shaking up; fourthly, water bath is carried out for 30min at the temperature of 90 ℃; cold water bath for 10 min; the absorbance at 490nm of the UV was measured and the results are shown in Table 1.
TABLE 1
| Absorbance of the solution | Concentration/. mu.g/mL |
| 0.1398 | 10.44 |
| 0.2946 | 20.88 |
| 0.6564 | 52.20 |
| 1.0896 | 83.52 |
| 1.275 | 104.40 |
Drawing a standard curve according to the data in the table 1As shown in fig. 2, where y is 81.491x-2.0287(y is concentration, x is absorbance value, unit μ g/mL); r2＝0.9969。
(2) 0.01025g, 0.00988g, 0.01017g and 0.01525g of the ganoderma lucidum polysaccharide samples obtained by the preparation method after freeze drying are accurately weighed respectively and dissolved in 200mL of purified water to obtain 51.25 mu g/mL, 49.40 mu g/mL, 50.85 mu g/mL and 76.25 mu g/mL ganoderma lucidum polysaccharide solutions.
(3) Taking 2mL of sample and 2mL of water respectively, measuring polysaccharide by a sulfuric acid phenol method, taking water as blank, measuring the absorbance at 490nm to obtain 0.7011, 0.6785, 0.6979 and 0.9590, substituting y for 81.491x-2.0287 to obtain 55.10 mu g/mL, 53.26 mu g/mL, 54.84 mu g/mL and 76.12 mu g/mL, and calculating the purity to be 107.51%, 107.81%, 107.85% and 99.83% (by adopting the measuring method, the obtained purity is 111.11% in theory).
In conclusion, the purity of the ganoderma lucidum polysaccharide obtained by the preparation method provided by the invention is up to more than 99%, and the ganoderma lucidum polysaccharide can be directly used in various ways such as experiments, medicine, production and the like.
It should be noted that the specific embodiments are merely representative examples of the present invention, and it is obvious that the technical solution of the present invention is not limited to the above-mentioned examples, and many variations are possible. Those skilled in the art, having the benefit of this disclosure and the benefit of this written description, will appreciate that other embodiments can be devised which do not depart from the specific details disclosed herein.
Claims (7)
1. A preparation process of high-purity ganoderma lucidum polysaccharide is characterized by comprising the following steps:
(1) firstly preparing a fermentation culture medium, and then culturing in a fermentation tank under the culture conditions of: inoculating Ganoderma mycelia 10%, fermenting at 29 deg.C, stirring at 150rpm/min, fermenting at pH5.7 under 0.05Mpa, and fermenting to obtain Ganoderma fermentation liquid;
(2) centrifuging the ganoderma lucidum fermentation liquor obtained in the step (2) to collect mycelia, adding purified water, uniformly stirring, centrifuging to collect ganoderma lucidum mycelia, and repeating the step for three times;
(3) adding purified water into the ganoderma lucidum mycelia collected in the step (2), boiling for 2h, centrifuging to collect ganoderma lucidum mycelia, and repeating for three times to obtain clear and transparent supernatant;
(4) adding purified water into the ganoderma lucidum mycelia after the impurities are removed in the step (3), mechanically crushing the ganoderma lucidum mycelia in a wall breaking machine for 2min, adding four times of volume of purified water, boiling for 2h, centrifuging, removing the precipitate, and taking supernatant; adding ethanol with the same volume, precipitating with ethanol for 8h, centrifuging to remove precipitate, boiling with purified water for dissolving, centrifuging, and collecting supernatant; precipitating with ethanol, re-dissolving, centrifuging, collecting supernatant for three times to obtain pure ganoderan, and oven drying or lyophilizing to obtain high-purity ganoderan dry powder.
2. The process for preparing high-purity ganoderma lucidum polysaccharide according to claim 1, wherein the fermentation medium in the step (1) comprises the following components: 3 percent of glucose, 1 percent of yeast powder, 0.4 percent of monopotassium phosphate, 0.4 percent of magnesium sulfate, 0.2 thousandth of defoaming agent and the balance of water.
3. The process according to claim 1, wherein the fermentation period of step (1) is 4-5 days.
4. The process according to claim 1, wherein the rotation speed of the centrifugation in step (2) is 4000 r/min.
5. The process according to claim 1, wherein the volume ratio of the purified water to the ganoderma lucidum mycelia in the step (3) is 5: 1.
6. the process according to claim 1, wherein the boiling step (3) is carried out for 2 hours.
7. The process for preparing high-purity ganoderma lucidum polysaccharide according to claim 1, wherein the rotating speed of the mechanical crushing of the wall breaking machine in the step (4) is 25000 r/min.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110538939.3A CN113151372B (en) | 2021-05-18 | 2021-05-18 | Preparation process of high-purity ganoderma lucidum polysaccharide |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110538939.3A CN113151372B (en) | 2021-05-18 | 2021-05-18 | Preparation process of high-purity ganoderma lucidum polysaccharide |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN113151372A true CN113151372A (en) | 2021-07-23 |
| CN113151372B CN113151372B (en) | 2022-09-16 |
Family
ID=76876230
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110538939.3A Active CN113151372B (en) | 2021-05-18 | 2021-05-18 | Preparation process of high-purity ganoderma lucidum polysaccharide |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113151372B (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115074406A (en) * | 2022-07-22 | 2022-09-20 | 上海市农业科学院 | Ganoderma lucidum liquid submerged fermentation extracellular fluid and fermentation method and application thereof |
| CN115634241A (en) * | 2022-11-16 | 2023-01-24 | 青岛农业大学 | Preparation method of Shandong ganoderma lucidum extract and application of Shandong ganoderma lucidum extract in preparation of hypoglycemic drugs |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102977222A (en) * | 2012-12-20 | 2013-03-20 | 西藏金稞集团有限责任公司 | Method for extracting ganoderma lucidum mycelia polysaccharide germanium |
| CN104059162A (en) * | 2014-06-25 | 2014-09-24 | 上海市农业科学院 | Ganoderma lucidum polysaccharide and preparation method thereof |
| CN105367675A (en) * | 2015-11-20 | 2016-03-02 | 临沂大学 | Method for extraction of polysaccharide from ganoderma lucidum karst |
| CN108530549A (en) * | 2018-04-12 | 2018-09-14 | 湖南御家化妆品制造有限公司 | Method for extracting polysaccharide from ganoderma lucidum mycelia |
-
2021
- 2021-05-18 CN CN202110538939.3A patent/CN113151372B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102977222A (en) * | 2012-12-20 | 2013-03-20 | 西藏金稞集团有限责任公司 | Method for extracting ganoderma lucidum mycelia polysaccharide germanium |
| CN104059162A (en) * | 2014-06-25 | 2014-09-24 | 上海市农业科学院 | Ganoderma lucidum polysaccharide and preparation method thereof |
| CN105367675A (en) * | 2015-11-20 | 2016-03-02 | 临沂大学 | Method for extraction of polysaccharide from ganoderma lucidum karst |
| CN108530549A (en) * | 2018-04-12 | 2018-09-14 | 湖南御家化妆品制造有限公司 | Method for extracting polysaccharide from ganoderma lucidum mycelia |
Non-Patent Citations (1)
| Title |
|---|
| 黄晓德等: "芦笋茎叶多糖的提取纯化研究", 《江西农业学报》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115074406A (en) * | 2022-07-22 | 2022-09-20 | 上海市农业科学院 | Ganoderma lucidum liquid submerged fermentation extracellular fluid and fermentation method and application thereof |
| CN115074406B (en) * | 2022-07-22 | 2024-01-05 | 上海市农业科学院 | Ganoderma lucidum liquid deep fermentation extracellular fluid and fermentation method and application thereof |
| CN115634241A (en) * | 2022-11-16 | 2023-01-24 | 青岛农业大学 | Preparation method of Shandong ganoderma lucidum extract and application of Shandong ganoderma lucidum extract in preparation of hypoglycemic drugs |
Also Published As
| Publication number | Publication date |
|---|---|
| CN113151372B (en) | 2022-09-16 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN113151372B (en) | Preparation process of high-purity ganoderma lucidum polysaccharide | |
| CN110041172A (en) | A kind of technique improving cannabidiol recovery rate using microbiological treatment hemp floral leaf | |
| US9284585B2 (en) | Method for increasing yield of total flavonoids in Ganoderma lucidum mycelium | |
| CN106636288B (en) | Method for extracting tigogenin by fermentation | |
| CN112972520B (en) | Method for improving active ingredient yield by deeply fermenting eucommia ulmoides leaves with inonotus obliquus liquid | |
| CN106929427B (en) | Culture medium suitable for high-yield extracellular polysaccharide of medicinal fungus ganoderma lucidum and culture method thereof | |
| CN101492706A (en) | Method for improving cordyceps sinensis bacterium native volume of production with cordyceps militaris link liquid fermentation | |
| CN113121557A (en) | Marine fungus-derived heteroterpene compound and application thereof in preparation of anti-inflammatory drugs | |
| CN111500472B (en) | Cordyceps gunnii mycelium rich in flavone and polyphenol and production method thereof | |
| CN112322508A (en) | Ganoderma lucidum mycelium culture method for improving content of ganoderma lucidum polysaccharide | |
| CN111635916A (en) | Pretreatment method for yeast beta-glucan extraction | |
| CN111533823A (en) | Process for extracting polysaccharide from ganoderma lucidum mycelia | |
| CN110551234A (en) | preparation method of pueraria polysaccharide and application of pueraria polysaccharide as growth promoter | |
| CN110846354B (en) | Preparation method of blackberry lily isoflavone aglycone | |
| CN109504725B (en) | Method for preparing high-purity hericium erinaceus polysaccharide by fermenting hericium erinaceus and fermentation culture medium | |
| CN111647092B (en) | Method for improving pachyman yield by utilizing semi-solid fermentation of trichoderma reesei and application of method | |
| CN110833560B (en) | Application of 2,4, 6-tri-O-galloyl-D-glucose in decocting of Chinese herbal medicines in preparation of antitumor medicines | |
| CN111118083A (en) | Method for extracting ganoderma lucidum polysaccharide | |
| CN109182148B (en) | Method for increasing cordycepic acid content in cordyceps militaris mycelia | |
| CN111440249A (en) | Method for extracting Fuzhuan tea polysaccharide | |
| CN108059686B (en) | Method for preparing polysaccharide based on pummelo peel | |
| CN103396295B (en) | A kind of method extracting resveratrol ingredient from giant knotweed | |
| CN111647511A (en) | Combined enzymolysis wall-breaking process for lucid ganoderma | |
| CN111349097A (en) | Preparation method of matrine | |
| CN114317282B (en) | Liquid fermentation method of Antrodia camphorata |
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 | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| PE01 | Entry into force of the registration of the contract for pledge of patent right |
Denomination of invention: Preparation of a high purity Ganoderma lucidum polysaccharide Effective date of registration: 20221212 Granted publication date: 20220916 Pledgee: Qingdao Jinling Shengtai Financing Guarantee Co.,Ltd. Pledgor: QINGDAO RUNDA BIOTECHNOLOGY Co.,Ltd. Registration number: Y2022370010165 |
|
| PE01 | Entry into force of the registration of the contract for pledge of patent right | ||
| PC01 | Cancellation of the registration of the contract for pledge of patent right |
Granted publication date: 20220916 Pledgee: Qingdao Jinling Shengtai Financing Guarantee Co.,Ltd. Pledgor: QINGDAO RUNDA BIOTECHNOLOGY Co.,Ltd. Registration number: Y2022370010165 |
|
| PE01 | Entry into force of the registration of the contract for pledge of patent right |
Denomination of invention: Preparation process of a high-purity Ganoderma lucidum polysaccharide Granted publication date: 20220916 Pledgee: China Construction Bank Co.,Ltd. Jimo Branch Pledgor: QINGDAO RUNDA BIOTECHNOLOGY Co.,Ltd. Registration number: Y2024370010065 |
|
| PE01 | Entry into force of the registration of the contract for pledge of patent right |