CN116535536A - Extraction method of Phellinus linteus polysaccharide - Google Patents
Extraction method of Phellinus linteus polysaccharide Download PDFInfo
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- CN116535536A CN116535536A CN202310704561.9A CN202310704561A CN116535536A CN 116535536 A CN116535536 A CN 116535536A CN 202310704561 A CN202310704561 A CN 202310704561A CN 116535536 A CN116535536 A CN 116535536A
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- phellinus linteus
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- linteus polysaccharide
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- 241000001727 Tropicoporus linteus Species 0.000 title claims abstract description 114
- 150000004676 glycans Chemical class 0.000 title claims abstract description 91
- 229920001282 polysaccharide Polymers 0.000 title claims abstract description 91
- 239000005017 polysaccharide Substances 0.000 title claims abstract description 91
- 238000000605 extraction Methods 0.000 title claims abstract description 19
- 239000002904 solvent Substances 0.000 claims abstract description 44
- 230000005496 eutectics Effects 0.000 claims abstract description 40
- 108090000790 Enzymes Proteins 0.000 claims abstract description 39
- 102000004190 Enzymes Human genes 0.000 claims abstract description 39
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 30
- 238000011282 treatment Methods 0.000 claims abstract description 26
- 238000000034 method Methods 0.000 claims abstract description 22
- 239000000243 solution Substances 0.000 claims abstract description 19
- 239000000843 powder Substances 0.000 claims abstract description 14
- 238000001914 filtration Methods 0.000 claims abstract description 11
- 239000011259 mixed solution Substances 0.000 claims abstract description 10
- 238000010298 pulverizing process Methods 0.000 claims abstract description 7
- 238000001816 cooling Methods 0.000 claims abstract description 6
- 238000007873 sieving Methods 0.000 claims abstract description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 6
- 238000002156 mixing Methods 0.000 claims abstract description 5
- 238000006243 chemical reaction Methods 0.000 claims abstract description 3
- 238000001035 drying Methods 0.000 claims abstract description 3
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims description 17
- 239000001257 hydrogen Substances 0.000 claims description 12
- 229910052739 hydrogen Inorganic materials 0.000 claims description 12
- KWIUHFFTVRNATP-UHFFFAOYSA-N Betaine Natural products C[N+](C)(C)CC([O-])=O KWIUHFFTVRNATP-UHFFFAOYSA-N 0.000 claims description 10
- KWIUHFFTVRNATP-UHFFFAOYSA-O N,N,N-trimethylglycinium Chemical compound C[N+](C)(C)CC(O)=O KWIUHFFTVRNATP-UHFFFAOYSA-O 0.000 claims description 10
- 229960003237 betaine Drugs 0.000 claims description 10
- 239000001763 2-hydroxyethyl(trimethyl)azanium Substances 0.000 claims description 8
- 235000019743 Choline chloride Nutrition 0.000 claims description 8
- SGMZJAMFUVOLNK-UHFFFAOYSA-M choline chloride Chemical compound [Cl-].C[N+](C)(C)CCO SGMZJAMFUVOLNK-UHFFFAOYSA-M 0.000 claims description 8
- 229960003178 choline chloride Drugs 0.000 claims description 8
- 239000007788 liquid Substances 0.000 claims description 7
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 6
- 239000001913 cellulose Substances 0.000 claims description 6
- 229920002678 cellulose Polymers 0.000 claims description 6
- 108010029541 Laccase Proteins 0.000 claims description 4
- 102000003992 Peroxidases Human genes 0.000 claims description 4
- 125000002887 hydroxy group Chemical group [H]O* 0.000 claims description 4
- 108040007629 peroxidase activity proteins Proteins 0.000 claims description 4
- 230000035484 reaction time Effects 0.000 claims description 3
- FBPFZTCFMRRESA-FSIIMWSLSA-N D-Glucitol Natural products OC[C@H](O)[C@H](O)[C@@H](O)[C@H](O)CO FBPFZTCFMRRESA-FSIIMWSLSA-N 0.000 claims description 2
- FBPFZTCFMRRESA-JGWLITMVSA-N D-glucitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-JGWLITMVSA-N 0.000 claims description 2
- 102000016680 Dioxygenases Human genes 0.000 claims description 2
- 108010028143 Dioxygenases Proteins 0.000 claims description 2
- 102000005744 Glycoside Hydrolases Human genes 0.000 claims description 2
- 108010031186 Glycoside Hydrolases Proteins 0.000 claims description 2
- PWHULOQIROXLJO-UHFFFAOYSA-N Manganese Chemical compound [Mn] PWHULOQIROXLJO-UHFFFAOYSA-N 0.000 claims description 2
- TVXBFESIOXBWNM-UHFFFAOYSA-N Xylitol Natural products OCCC(O)C(O)C(O)CCO TVXBFESIOXBWNM-UHFFFAOYSA-N 0.000 claims description 2
- CDQSJQSWAWPGKG-UHFFFAOYSA-N butane-1,1-diol Chemical compound CCCC(O)O CDQSJQSWAWPGKG-UHFFFAOYSA-N 0.000 claims description 2
- 229910052748 manganese Inorganic materials 0.000 claims description 2
- 239000011572 manganese Substances 0.000 claims description 2
- HEBKCHPVOIAQTA-UHFFFAOYSA-N meso ribitol Natural products OCC(O)C(O)C(O)CO HEBKCHPVOIAQTA-UHFFFAOYSA-N 0.000 claims description 2
- 239000000600 sorbitol Substances 0.000 claims description 2
- 239000000811 xylitol Substances 0.000 claims description 2
- HEBKCHPVOIAQTA-SCDXWVJYSA-N xylitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)CO HEBKCHPVOIAQTA-SCDXWVJYSA-N 0.000 claims description 2
- 229960002675 xylitol Drugs 0.000 claims description 2
- 235000010447 xylitol Nutrition 0.000 claims description 2
- 239000000126 substance Substances 0.000 description 10
- 230000000694 effects Effects 0.000 description 9
- 229920005610 lignin Polymers 0.000 description 6
- LYCAIKOWRPUZTN-UHFFFAOYSA-N ethylene glycol Natural products OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 description 5
- 230000009471 action Effects 0.000 description 4
- 230000000052 comparative effect Effects 0.000 description 4
- 238000004090 dissolution Methods 0.000 description 4
- 239000002244 precipitate Substances 0.000 description 4
- 239000000370 acceptor Substances 0.000 description 3
- 238000004880 explosion Methods 0.000 description 3
- 239000012535 impurity Substances 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 description 2
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 2
- 239000004472 Lysine Substances 0.000 description 2
- 241000789545 Trimeria grandifolia Species 0.000 description 2
- 229920002522 Wood fibre Polymers 0.000 description 2
- 238000010170 biological method Methods 0.000 description 2
- 238000000354 decomposition reaction Methods 0.000 description 2
- 229930003935 flavonoid Natural products 0.000 description 2
- 235000017173 flavonoids Nutrition 0.000 description 2
- 150000002215 flavonoids Chemical class 0.000 description 2
- 235000013922 glutamic acid Nutrition 0.000 description 2
- 239000004220 glutamic acid Substances 0.000 description 2
- 229920002521 macromolecule Polymers 0.000 description 2
- 238000001556 precipitation Methods 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 230000002195 synergetic effect Effects 0.000 description 2
- 150000003648 triterpenes Chemical class 0.000 description 2
- 238000003809 water extraction Methods 0.000 description 2
- 239000002025 wood fiber Substances 0.000 description 2
- 206010012735 Diarrhoea Diseases 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 241000123113 Phellinus igniarius Species 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 229930013930 alkaloid Natural products 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000003110 anti-inflammatory effect Effects 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 230000000740 bleeding effect Effects 0.000 description 1
- 230000017531 blood circulation Effects 0.000 description 1
- 235000010633 broth Nutrition 0.000 description 1
- 210000002421 cell wall Anatomy 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 238000011097 chromatography purification Methods 0.000 description 1
- 239000002131 composite material Substances 0.000 description 1
- 238000004185 countercurrent chromatography Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000000593 degrading effect Effects 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 238000010262 high-speed countercurrent chromatography Methods 0.000 description 1
- 230000002218 hypoglycaemic effect Effects 0.000 description 1
- 230000004957 immunoregulator effect Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 238000002386 leaching Methods 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 238000000643 oven drying Methods 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- 238000000053 physical method Methods 0.000 description 1
- 150000008442 polyphenolic compounds Chemical class 0.000 description 1
- 235000013824 polyphenols Nutrition 0.000 description 1
- 230000001376 precipitating effect Effects 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 150000003431 steroids Chemical class 0.000 description 1
- 238000002604 ultrasonography Methods 0.000 description 1
- 238000002137 ultrasound extraction 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
- C08B37/0003—General processes for their isolation or fractionation, e.g. purification or extraction from biomass
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- General Health & Medical Sciences (AREA)
- Sustainable Development (AREA)
- Biochemistry (AREA)
- Materials Engineering (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Medicinal Chemistry (AREA)
- Polymers & Plastics (AREA)
- Organic Chemistry (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention discloses a method for extracting Phellinus linteus polysaccharide, which comprises the following steps: pulverizing Phellinus linteus fruiting body, and sieving with 40-60 mesh sieve; adding Phellinus linteus fruiting body powder into eutectic solvent containing biological enzyme for reaction to obtain mixed solution; filtering and concentrating the mixed solution to obtain a concentrated solution, adding an ethanol solution into the concentrated solution, and standing for 6-12 h to obtain crude Phellinus linteus polysaccharide; mixing crude Phellinus linteus polysaccharide with water, performing microwave treatment, cooling to room temperature, centrifuging, filtering, and drying to obtain Phellinus linteus polysaccharide. The extraction method is simple and efficient, and has high Phellinus linteus polysaccharide yield.
Description
Technical Field
The invention belongs to the technical field of traditional Chinese medicine extraction, and particularly relates to an extraction method of Phellinus linteus polysaccharide.
Background
Phellinus linteus (Phellinus Igniarius), also known as Phellinus linteus, is a precious perennial large-scale medicinal fungus. Sang Huangshi the medicinal theory has the effects of promoting blood circulation, stopping bleeding, resolving food stagnation, and relieving diarrhea. It is counted that Phellinus linteus is mainly distributed in korea, southeast asia, australia, america, etc. abroad, and domestic in northeast, northwest, southwest, etc. Phellinus linteus contains polysaccharides, flavonoids, triterpenes, polyphenols, steroids, pyrones and alkaloids, wherein polysaccharides, flavonoids and triterpenes are main functional active ingredients of Phellinus linteus. Phellinus linteus has been attracting great attention because of its remarkable pharmacological functions of anti-tumor, antioxidant, immunoregulatory, anti-inflammatory and hypoglycemic. Phellinus linteus polysaccharides can be present in Phellinus linteus fruiting bodies, fermentation broths and mycelia. The wild mulberry Huang Shengchang has a long period and takes a long time to grow into a size suitable for medical use, and the wild mulberry Huang Xishao has a huge market demand and is expensive in recent years.
However, phellinus linteus fruiting body is hard and lignified, and the cell wall contains a large amount of cellulose macromolecules, and its functional components are usually linked with macromolecules such as cellulose and lignin, which makes extraction of Phellinus linteus polysaccharide difficult.
At present, the existing Phellinus linteus polysaccharide extraction technology has more defects and drawbacks, mainly comprises water extraction and alcohol precipitation, and has lower extraction efficiency and longer extraction time. Pretreatment is usually carried out by physical, chemical or biological technology to assist water extraction and alcohol precipitation, although these modes achieve certain effects. However, these techniques often have the disadvantages of long reaction time, high cost, incomplete and unstable polysaccharide extraction, etc., which is disadvantageous for popularization and application of Yu Sanghuang polysaccharide.
Chinese patent publication No. CN101323648A discloses a method for extracting crude Phellinus linteus polysaccharide and purifying Phellinus linteus polysaccharide. The technical problem to be solved in the patent is to disclose an extraction method of a Phellinus linteus crude polysaccharide extract and a method for preparing Phellinus linteus polysaccharide by high-efficiency countercurrent chromatography. The patent uses ultrasonic extraction method to extract Phellinus linteus crude polysaccharide from Phellinus linteus fruiting body, and uses high-speed countercurrent chromatography to further purify Phellinus linteus polysaccharide. But involves chromatographic purification techniques at a relatively high cost.
Chinese patent publication No. CN 114044832A discloses a method for extracting Phellinus linteus fruiting body polysaccharide; pulverizing dried Phellinus linteus fruiting body by pulverizer, sieving, putting into steam explosion device to complete explosion, collecting steam exploded sample, placing into oven, oven drying, pulverizing, precipitating with ethanol, redissolving the precipitate, and respectively treating with microwave and ultrasound to obtain Phellinus linteus fruiting body polysaccharide. The steam explosion pretreatment and the microwave and ultrasonic combined treatment further promote the leaching of Phellinus linteus fruiting body substances, the yield reaches 8.1 percent, the industrial cost is low, the production cycle time is short, the integrity of polysaccharide is ensured, and the Phellinus linteus fruiting body substances are safe, green and nontoxic. But the method has complicated steps, more related equipment and higher cost.
In the prior art, a physical method is generally adopted, and a biological method is adopted for extracting Phellinus linteus crude polysaccharide, so that the adopted method is complex, the cost is high, the extraction process can be simplified, impurities can be removed accurately and efficiently, and the required Phellinus linteus polysaccharide is one of the problems to be solved in the field.
Disclosure of Invention
The invention provides a Phellinus linteus polysaccharide extraction method which is simple and efficient and has higher Phellinus linteus polysaccharide yield.
The embodiment of the invention provides an extraction method of Phellinus linteus polysaccharide, which comprises the following steps:
(1) Pulverizing Phellinus linteus fruiting body, and sieving with 40-60 mesh sieve;
(2) Adding the Phellinus linteus fruiting body powder obtained in the step (1) into a eutectic solvent containing biological enzyme to react to obtain a mixed solution;
(3) Filtering and concentrating the mixed solution obtained in the step (2) to obtain a concentrated solution, adding an ethanol solution into the concentrated solution, and standing for 6-12 h to obtain crude Phellinus linteus polysaccharide;
(4) Mixing the crude Phellinus linteus polysaccharide obtained in the step (3) with water, performing microwave treatment, cooling to room temperature, centrifuging, filtering, and drying to obtain Phellinus linteus polysaccharide.
According to the invention, lignin of the Phellinus linteus fruiting body powder is dissolved and destroyed by the biological enzyme and the eutectic solvent, so that Phellinus linteus polysaccharide can be dissolved out easily, and under the action of microwaves, phellinus linteus polysaccharide substances can be melted rapidly and efficiently, thereby avoiding the generation of impurities and having higher yield.
Further, the eutectic solvent comprises a hydrogen bond donor and a hydrogen bond acceptor, wherein the hydrogen bond acceptor comprises choline chloride and/or betaine, and the donor is a solvent containing hydroxyl hydrogen bonds.
Further, the solvent containing hydroxyl hydrogen bonds is one or more of methanol, ethanol, butanediol, glycerol, xylitol and sorbitol.
The invention provides the eutectic solvent with proper ingredients, so that the active enzyme can keep activity under the eutectic solvent, the connection between lignin and Phellinus linteus polysaccharide can be accurately destroyed by using the active enzyme, the lignin is dissolved by the eutectic solvent, and the eutectic solvent provided by the invention can also avoid the decomposition of Phellinus linteus polysaccharide and reduce the generation of impurities.
Further, the biological enzyme is one or more of laccase, peroxidase, cellulose complex enzyme, glycosidase, manganese-based peroxidase and dioxygenase.
The biological enzyme provided by the invention can destroy the connection between lignin and Phellinus linteus polysaccharide, and can avoid decomposition of Phellinus linteus polysaccharide, so that higher Phellinus linteus polysaccharide yield can be obtained.
Further preferably, after pulverizing Phellinus linteus fruiting body, sieving with 60 mesh sieve.
Further, the solid-to-liquid ratio of the Phellinus linteus fruiting body powder to the eutectic solvent is 1:10-1:40 g/mL.
Further, the addition amount of the biological enzyme is 0.5% -5% of the mass of the Phellinus linteus fruiting body powder.
Further, the Phellinus linteus fruiting body powder is added into the eutectic solvent containing biological enzyme to react to obtain mixed solution, the reaction temperature is 35 ℃ to 55 ℃, and the reaction time is 24 hours to 48 hours.
Further, the ethanol solution is 80% -100% ethanol solution, and the volume ratio of the ethanol solution to the mixed solution is 1:10-1:30.
Further, the power of the microwave treatment is 100-400 w, and the microwave treatment time is 2-8 min.
The lignin of the Phellinus linteus fruiting body powder is removed and dissolved by adding the eutectic solvent containing biological enzyme to obtain crude Phellinus linteus polysaccharide, so that the Phellinus linteus polysaccharide is extracted by utilizing proper microwave treatment parameters under the premise of lower power, poorer extraction effect and higher power to destroy the Phellinus linteus polysaccharide, so that the Phellinus linteus polysaccharide is decomposed.
Compared with the prior art, the invention has the beneficial effects that:
(1) The eutectic solvent and the biological enzyme are cooperated to break the connection bond in the wood fiber, promote the entry and attack of the eutectic solvent containing the enzyme preparation, break the connection between the polysaccharide and the woody substances under the accurate action of the biological enzyme, dissolve the lignified part into the eutectic solvent, promote the dissolution of the polysaccharide substances, and rapidly and efficiently dissolve the polysaccharide substances under the action of microwaves. The invention comprehensively considers the physical and chemical treatment strength and the process condition, and realizes the efficient dissolution of the polysaccharide in the Phellinus linteus fruiting body.
(2) The biological enzyme in the eutectic solvent can keep activity, so that the sample is accurately treated, and the dissolution yield of polysaccharide is improved; the microwave with smaller energy is adopted for post-treatment, and on the premise of ensuring the activity and minimum loss of polysaccharide, the Phellinus linteus polysaccharide is efficiently dissolved, and the highest yield reaches 9.8 percent.
Drawings
FIG. 1 is a bar graph showing the effect of various microwave treatments provided in examples 10-14 on Phellinus linteus polysaccharide yields;
FIG. 2 is a bar graph of the yields of Phellinus linteus polysaccharide extracted in example 1 and comparative examples 1-3.
Detailed Description
The present invention will be further described with reference to specific examples, but the invention is not limited thereto, and other equivalents and modifications may be made thereto without departing from the spirit and scope of the present invention, and are intended to fall within the scope of the present invention.
Example 1
The embodiment provides a method for extracting Phellinus linteus polysaccharide based on biological enzyme synergistic eutectic solvent, which comprises the following steps:
pulverizing Phellinus linteus fruiting body, and sieving with 60 mesh sieve to obtain Phellinus linteus fruiting body powder.
100g of Phellinus linteus fruiting body powder is added into eutectic solvent containing biological enzyme. The eutectic solvent consists of choline chloride and glycerol; the biological enzyme is laccase and cellulose composite enzyme, and the eutectic solvent and the biological enzyme are mixed according to the mass ratio of 1:1;
the solid-to-liquid ratio of the crushed and sieved Phellinus linteus fruiting body powder to the eutectic solvent containing biological enzyme is 1:30g/mL, the treatment temperature of the eutectic solvent containing biological enzyme is 45 ℃, the treatment time is 48h, and the adding amount of biological enzyme is 1.5% of the mass of the crushed fruiting body.
After the treatment of the eutectic solvent containing biological enzyme is finished, gauze filtration separation and concentration are carried out, 80% ethanol solution is added into the obtained concentrated solution, the mixture is added according to the volume ratio of 1:10, the mixture is stood for 12 hours, and centrifugal separation is carried out, thus obtaining the precipitate, namely crude Phellinus linteus polysaccharide.
Mixing the obtained precipitate with water according to a solid-to-liquid ratio of 1:30, treating with microwave for 2min under the condition of microwave power of 200W, cooling to room temperature, centrifuging, filtering, and concentrating to obtain Phellinus linteus polysaccharide with a yield of 9.8%.
Example 2-example 9
Preparing different types of eutectic solvents, wherein the compositions are shown in table 1, treating for 48 hours at 45 ℃ according to the solid-to-liquid ratio of 1:30g/mL of crushed and sieved Phellinus linteus fruiting bodies, adding biological enzyme in an amount of 1.5% of the mass of the Phellinus linteus fruiting bodies, filtering and concentrating, adding 80% ethanol solution into the obtained concentrated solution, adding according to the volume ratio of 1:10, standing for 12 hours, and centrifuging. Mixing the obtained precipitate with water according to a solid-to-liquid ratio of 1:30, treating with microwave under microwave power of 200W for 2min, cooling to room temperature, centrifuging, filtering, and concentrating to obtain Phellinus linteus polysaccharide. Results of polysaccharide extraction rates with different eutectic solvents.
TABLE 1 correlation between different eutectic solvent species and Phellinus linteus polysaccharide yields
| Sequence number | Hydrogen bond acceptors | Hydrogen bond donors | Phellinus linteus polysaccharide yield% |
| Example 2 | Choline chloride | Ethylene glycol | 8.1 |
| Example 3 | Choline chloride | Glycerol | 9.8 |
| Example 4 | Choline chloride | Lysine | 7.1 |
| Example 5 | Choline chloride | Glutamic acid | 6.8 |
| Example 6 | Betaine (betaine) | Ethylene glycol | 5.8 |
| Example 7 | Betaine (betaine) | Glycerol | 6.9 |
| Example 8 | Betaine (betaine) | Lysine | 5.1 |
| Example 9 | Betaine (betaine) | Glutamic acid | 5.8 |
Example 10-example 14
Different from example 1, under the same treatment conditions of the eutectic solvent and the biological enzyme, the treatment is carried out for 2min by adopting different microwave powers, the solid-liquid ratio is 1:30g/mL, the microwave powers are selected to be 100W, 200W, 400W, 600W and 800W, the cooling is carried out to room temperature, and the Phellinus linteus fruiting body polysaccharide is obtained through centrifugation, filtration and concentration.
TABLE 2 relationship between different powers and Phellinus linteus polysaccharide yields
| Sequence number | Power of | Phellinus linteus polysaccharide yield% |
| Example 10 | 100W | 7.1 |
| Example 11 | 200W | 9.8 |
| Example 12 | 400W | 8.3 |
| Example 13 | 600W | 7.5 |
| Example 14 | 800W | 6.8 |
Comparative example 1
Unlike example 1, the eutectic solvent containing the bioenzyme was replaced with a eutectic solvent composed of choline chloride and glycerol, and the Phellinus linteus polysaccharide yield was only 4.3%.
Comparative example 2
Unlike example 1, the eutectic solvent containing the biological enzyme was replaced with the biological enzyme, which is a laccase and cellulose complex enzyme, and the Phellinus linteus polysaccharide yield was only 3.1%. The eutectic solvent and biological enzyme synergistic treatment has obvious promotion effect on the extraction of Phellinus linteus polysaccharide.
Comparative example 3
Unlike example 3, the eutectic solvent containing biological enzyme is not added, the crushed and sieved Phellinus linteus fruiting body is directly mixed with water for microwave treatment, the Phellinus linteus polysaccharide yield is 1.5%, which means that the treatment is carried out firstly and then the microwave treatment is carried out, the improvement of polysaccharide yield is more facilitated, the microwave effect of the treatment without the eutectic solvent is smaller, and the microwave action sites are improved due to the damage of the eutectic solvent and the biological enzyme to the network structure and the connecting bond of the wood fiber, thereby facilitating the extraction of Phellinus linteus polysaccharide.
Performance analysis:
as shown in figure 1, the yield of Phellinus linteus polysaccharide under different microwave treatment conditions is different, and the yield of Phellinus linteus polysaccharide is increased and reduced along with the increase of microwave power, and is 9.8% at the maximum of 200W, because the woody substances in Phellinus linteus fruiting body are dissolved out under the previous treatment conditions, the structure is loose, the proper microwave power is favorable for the dissolution of Phellinus linteus polysaccharide, however, when the microwave power is too high, the Phellinus linteus polysaccharide is degraded or destroyed, and the yield of Phellinus linteus polysaccharide is further reduced. The method has the advantage that the power requirement is obviously lower than that of a similar method because the method is subjected to the treatment of the eutectic solvent of the biological enzyme in the early stage.
As shown in figure 2, the yields of Phellinus linteus polysaccharide by using only eutectic solvents, biological enzymes and microwave processors are respectively 4.3%,3.1% and 1.5%, and the yields of Phellinus linteus polysaccharide by using the processing steps alone are obviously lower than that of the Phellinus linteus polysaccharide by combining the steps, namely, under the common treatment of the eutectic solvents containing the biological enzymes and microwaves, the yields of Phellinus linteus polysaccharide can reach 9.8%, which shows that the physical, chemical and biological methods are organically combined, and the yields of Phellinus linteus polysaccharide can be promoted and improved on the premise of not damaging or degrading the Phellinus linteus polysaccharide.
Claims (9)
1. An extraction method of Phellinus linteus polysaccharide, comprising:
(1) Pulverizing Phellinus linteus fruiting body, and sieving with 40-60 mesh sieve;
(2) Adding the Phellinus linteus fruiting body powder obtained in the step (1) into a eutectic solvent containing biological enzyme to react to obtain a mixed solution;
(3) Filtering and concentrating the mixed solution obtained in the step (2) to obtain a concentrated solution, adding an ethanol solution into the concentrated solution, and standing for 6-12 h to obtain crude Phellinus linteus polysaccharide;
(4) Mixing the crude Phellinus linteus polysaccharide obtained in the step (3) with water, performing microwave treatment, cooling to room temperature, centrifuging, filtering, and drying to obtain Phellinus linteus polysaccharide.
2. The method for extracting Phellinus linteus polysaccharide as claimed in claim 1, wherein the eutectic solvent comprises hydrogen bond donor and hydrogen bond acceptor, the hydrogen bond acceptor comprises choline chloride and/or betaine, and the donor is a solvent containing hydroxyl hydrogen bonds.
3. The method for extracting Phellinus linteus polysaccharide as claimed in claim 2, wherein the solvent containing hydroxyl hydrogen bond is one or more of methanol, ethanol, butanediol, glycerol, xylitol, and sorbitol.
4. The method for extracting Phellinus linteus polysaccharide as claimed in claim 1, wherein the biological enzyme is one or more of laccase, peroxidase, cellulose complex enzyme, glycosidase, manganese-based peroxidase, dioxygenase.
5. The method for extracting Phellinus linteus polysaccharide as claimed in claim 1, wherein the solid-to-liquid ratio of Phellinus linteus fruiting body powder to eutectic solvent is 1:10-1:50 g/mL.
6. The method for extracting Phellinus linteus polysaccharide as claimed in claim 1, wherein the amount of the bio-enzyme added is 0.5-5% of the powder mass of Phellinus linteus fruiting body.
7. The method for extracting Phellinus linteus polysaccharide as claimed in claim 1, wherein Phellinus linteus fruiting body powder is added into eutectic solvent containing biological enzyme to react to obtain mixed solution, the reaction temperature is 25-65deg.C, and the reaction time is 12-96 h.
8. The method for extracting Phellinus linteus polysaccharide as claimed in claim 1, wherein the ethanol solution is 80% -100% ethanol solution, and the volume ratio of the ethanol solution to the mixed solution is 1:10-1:50.
9. The method for extracting Phellinus linteus polysaccharide as claimed in claim 1, wherein the microwave treatment power is 100-400 w, and the microwave treatment time is 0.5-10 min.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE69925916D1 (en) * | 1999-10-18 | 2005-07-28 | Korea Inst Sci & Tech | Immunostimulatory Phellinus ssp. Polysaccharide and use |
| CN114044832A (en) * | 2021-12-15 | 2022-02-15 | 中华全国供销合作总社济南果品研究院 | Method for extracting phellinus igniarius sporocarp polysaccharide |
| CN116217748A (en) * | 2023-03-01 | 2023-06-06 | 宁波御菌生物技术有限公司 | Method for extracting polysaccharide from Phellinus linteus fruiting body |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE69925916D1 (en) * | 1999-10-18 | 2005-07-28 | Korea Inst Sci & Tech | Immunostimulatory Phellinus ssp. Polysaccharide and use |
| CN114044832A (en) * | 2021-12-15 | 2022-02-15 | 中华全国供销合作总社济南果品研究院 | Method for extracting phellinus igniarius sporocarp polysaccharide |
| CN116217748A (en) * | 2023-03-01 | 2023-06-06 | 宁波御菌生物技术有限公司 | Method for extracting polysaccharide from Phellinus linteus fruiting body |
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| 于秋菊: "复合酶辅助低共熔溶剂提取枸杞子多糖的工艺优化及活性研究", 食品科技, vol. 47, no. 14, pages 171 - 179 * |
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