阅读说明：本技术 一种灵芝菌丝体多糖的制备方法及其应用 (Preparation method and application of ganoderma lucidum mycelium polysaccharide ) 是由 王中振 谢涛 于 2020-06-08 设计创作，主要内容包括：本发明属于植物提取技术领域,具体涉及一种灵芝菌丝体多糖及其制备方法和应用。本发明采用含有丰富的灵芝多糖的灵芝菌丝体为提取原料,通过原料预处理、脱脂处理、超临界CO<Sub>2</Sub>提取、酶解、醇沉淀等工艺步骤制得灵芝菌丝体多糖。本发明提供的灵芝菌丝体多糖的制备方法制得的灵芝菌丝体多糖的提取率达到15.4％以上,纯度达到76.1％以上。本发明采用超临界CO<Sub>2</Sub>提取技术和酶解技术对灵芝菌丝体进行多糖提取,提高了多糖的提取率,避免了原材料的浪费,节约生产成本,制得的灵芝菌丝体多糖应用于保健品和药物的制备,具有良好的功效,经济效益和的和会效益显著。(The invention belongs to plant extractionThe technical field, in particular to a ganoderma lucidum mycelium polysaccharide and a preparation method and application thereof. The invention adopts ganoderma lucidum mycelium containing rich ganoderma lucidum polysaccharide as extraction raw material, and the ganoderma lucidum mycelium is subjected to pretreatment, degreasing treatment and supercritical CO 2 Extracting, performing enzymolysis, precipitating with ethanol, etc. to obtain Ganoderma mycelia polysaccharide. The extraction rate of the ganoderma lucidum mycelium polysaccharide prepared by the preparation method of the ganoderma lucidum mycelium polysaccharide provided by the invention reaches more than 15.4%, and the purity of the ganoderma lucidum mycelium polysaccharide reaches more than 76.1%. The invention adopts supercritical CO 2 The extraction technology and the enzymolysis technology carry out polysaccharide extraction on the ganoderma lucidum mycelia, the extraction rate of the polysaccharide is improved, the waste of raw materials is avoided, the production cost is saved, the prepared ganoderma lucidum mycelia polysaccharide is applied to the preparation of health-care products and medicines, the effect is good, and the economic benefit and the concordant benefit are obvious.)

1. The preparation method of the ganoderma lucidum mycelium polysaccharide is characterized by comprising the following steps:

s1, putting the dried ganoderma lucidum mycelia into a grinder to be ground to 8-10 mu m, then adding an organic solvent to carry out degreasing treatment, carrying out centrifugal separation, taking the lower-layer precipitate, and drying at 50-70 ℃ to obtain ganoderma lucidum mycelia coarse powder;

s2, adding 10-20 times of deionized water into the ganoderma lucidum mycelium coarse powder prepared in the step S1, and then performing supercritical CO₂Extracting, filtering to obtain water extract;

s3, taking the water extract prepared in the step S2, adding complex enzyme, and carrying out heat preservation enzymolysis under the stirring state of 400-500 rpm for 3-5 hours to prepare enzymolysis liquid;

s4, heating the enzymolysis liquid prepared in the step S3 to 100-120 ℃, preserving heat for 1 hour, naturally cooling to 20 ℃, and then performing spray drying to prepare polysaccharide coarse powder;

s5, adding 8-10 times of absolute ethyl alcohol into the polysaccharide coarse powder prepared in the step S4, stirring for 1-1.5 hours under the condition of 600-800 rpm, performing centrifugal separation, removing supernatant, and performing spray drying to obtain the polysaccharide coarse powder.

2. The method for preparing ganoderma lucidum mycelium polysaccharide according to claim 1, wherein the amount of the organic solvent added in the step S1 is 8-10 times of the mass of ganoderma lucidum mycelium; the organic solvent is a mixture of propylene glycol methyl ether, acetone and dimethyl sulfoxide; the degreasing time is 1-3 h.

3. The method for preparing ganoderma lucidum mycelium polysaccharide according to claim 2, wherein the mixing mass ratio of propylene glycol methyl ether, acetone and dimethyl sulfoxide is 2-3: 4-7: 1-2.

4. The method for preparing the ganoderma lucidum mycelium polysaccharide according to claim 1, wherein supercritical CO is adopted in the step S2₂The extraction conditions are as follows: the pressure is 30-35 Mpa, the extraction temperature is 50-60 ℃, and the extraction time is 4-5 h.

5. The preparation method of the ganoderma lucidum mycelium polysaccharide according to claim 1, wherein the complex enzyme in the step S3 is composed of cellulase, keratinase and hemicellulase according to a mass ratio of 5-7: 1-3: 3-5.

6. The method for preparing ganoderma lucidum mycelium polysaccharide according to claim 1, wherein the volume fraction of the absolute ethyl alcohol in the step S5 is 95%.

7. Use of the ganoderma lucidum mycelia polysaccharide prepared by the method for preparing ganoderma lucidum mycelia polysaccharide as claimed in claim 1 to 6 in health products and medicines.

8. The use of the ganoderma lucidum mycelia polysaccharides according to claim 7, wherein the medicament comprises tablets and oral liquids for treating immunoregulation, tumor resistance, aging resistance, cardiovascular and cerebrovascular diseases, blood pressure reduction, cough relieving and allergy resisting.

Technical Field

The invention belongs to the technical field of plant extraction, and particularly relates to ganoderma lucidum mycelium polysaccharide and a preparation method and application thereof.

Background

The ganoderma lucidum mycelium is hypha formed by germination of ganoderma lucidum spores, more than half of nutrient substances required by ganoderma lucidum sporocarp are derived from nutrients stored by weight of the hypha, so that the nutrient content of the mycelium and the content of ganoderma lucidum mycelium polysaccharide are generally many times higher than those of the ganoderma lucidum sporocarp and ganoderma lucidum spore powder.

The ganoderma lucidum mycelium polysaccharide is composed of three strands of monosaccharide chains, the configuration of the ganoderma lucidum mycelium polysaccharide is similar to that of DNA and RNA, the ganoderma lucidum mycelium polysaccharide is a spiral three-dimensional structure, and the spiral layers are mainly fixed by hydrogen bonds. The ganoderma lucidum mycelium polysaccharide has obvious differences in certain physicochemical properties such as monosaccharide composition, glycosidic bond configuration, molecular weight, optical rotation, solubility and the like. Most of more than 200 kinds of extracted ganoderma lucidum mycelium polysaccharides are heteropolysaccharides, namely, the polysaccharides contain other monosaccharides such as galactose, mannose, arabinose, xylose, fucose and rhamnose besides glucose; due to different sources, the molecular weight can vary from tens of thousands to hundreds of thousands, most of them are accompanied by branches, and part of the polysaccharide also contains peptide chains.

Ganoderan is one of the most effective components in ganoderma, and has the characteristics of stimulating host nonspecific resistance, immunospecific reaction and inhibiting tumor physiological activity. In addition, the ganoderma lucidum polysaccharide has good effects in the aspects of immunoregulation, tumor resistance, aging resistance, cardiovascular and cerebrovascular protection, liver protection, detoxification, blood pressure reduction, anaphylactic reaction reduction and the like. The ganoderma lucidum polysaccharide is the soul of ganoderma lucidum function, the content of ganoderma lucidum polysaccharide determines the efficacy of the product, and the ganoderma lucidum polysaccharide is the main component for treating various diseases.

Patent text with application number ZL201910800585.8 discloses a ganoderma lucidum polysaccharide extraction technology, and the extraction technology comprises the steps of raw material pretreatment, ganoderma lucidum crude polysaccharide extraction, ganoderma lucidum polysaccharide deproteinization, ganoderma lucidum extract purification, alcohol precipitation extract and the like, wherein the ganoderma lucidum crude polysaccharide extraction adopts hot water extraction, is assisted by ultrasonic extraction, and is further purified through the steps of enzymolysis of protein and alcohol precipitation, and finally the ganoderma lucidum polysaccharide is obtained. Although the extraction method can obtain a certain amount of ganoderan, the purity of ganoderan is not high, and the ganoderma is used as the extraction raw material, which causes a great amount of raw material waste in the extraction process, has high extraction cost, and is not suitable for industrial production.

In conclusion, the technical problems of low purity of ganoderma lucidum polysaccharide, serious waste of raw materials, unsuitability for industrial production and the like generally exist in the prior art.

Disclosure of Invention

In order to overcome the defects of the prior art, the invention aims to provide a preparation method and application of ganoderma lucidum mycelium polysaccharide. The ganoderma lucidum mycelium polysaccharide provided by the invention has high purity and good effect, directly adopts ganoderma lucidum mycelium with rich polysaccharide content as an extraction raw material, avoids the waste of the raw material, saves the cost, and is beneficial to realizing industrial production.

In order to achieve the purpose, the technical scheme of the invention is as follows:

a preparation method of ganoderma lucidum mycelium polysaccharide specifically comprises the following steps:

s1, putting the dried ganoderma lucidum mycelia into a grinder to be ground to 8-10 mu m, then adding an organic solvent to carry out degreasing treatment, carrying out centrifugal separation, taking the lower-layer precipitate, and drying at 50-70 ℃ to obtain ganoderma lucidum mycelia coarse powder;

s2, adding 10-20 times of deionized water into the ganoderma lucidum mycelium coarse powder prepared in the step S1, and then performing supercritical CO₂Extracting, filtering to obtain water extract;

s3, taking the water extract prepared in the step S2, adding complex enzyme, and carrying out heat preservation enzymolysis under the stirring state of 400-500 rpm for 3-5 hours to prepare enzymolysis liquid;

s4, heating the enzymolysis liquid prepared in the step S3 to 100-120 ℃, preserving heat for 1 hour, naturally cooling to 20 ℃, and then performing spray drying to prepare polysaccharide coarse powder;

s5, adding 8-10 times of absolute ethyl alcohol into the polysaccharide coarse powder prepared in the step S4, stirring for 1-1.5 hours under the condition of 600-800 rpm, performing centrifugal separation, removing supernatant, and performing spray drying to obtain the polysaccharide coarse powder.

Further, the adding amount of the organic solvent in the step S1 is 8-10 times of the mass of the ganoderma lucidum mycelia; the organic solvent is a mixture of propylene glycol methyl ether, acetone and dimethyl sulfoxide; the degreasing time is 1-3 h.

Furthermore, the mixing mass ratio of the propylene glycol methyl ether, the acetone and the dimethyl sulfoxide is 2-3: 4-7: 1-2.

Further, supercritical CO in step S2₂The extraction conditions are as follows: the pressure is 30-35 Mpa, the extraction temperature is 50-60 ℃, and the extraction time is 4-5 h.

Further, the complex enzyme in the step S3 is composed of cellulase, keratinase and hemicellulase according to a mass ratio of 5-7: 1-3: 3-5.

Further, the volume fraction of the absolute ethyl alcohol in the step S5 is 95%.

The invention also provides the application of the ganoderma lucidum mycelium polysaccharide prepared by the preparation method of the ganoderma lucidum mycelium polysaccharide in health care products and medicines.

Furthermore, the medicine comprises tablets and oral liquid for treating immunoregulation, tumor resistance, aging resistance, cardiovascular and cerebrovascular diseases, blood pressure reduction, cough relieving and allergy resistance.

The invention adopts the ganoderma lucidum mycelia as the extraction raw material of the ganoderma lucidum mycelia polysaccharide, effectively solves the problem of bitter taste of the fruiting body polysaccharide, and the prepared ganoderma lucidum mycelia polysaccharide has better taste when being applied to oral liquid. In addition, the fiber content of the ganoderma lucidum mycelia is low, the content of the basswood polysaccharide is high, and the content of the triterpene component in the ganoderma lucidum mycelia is low, so that the extraction rate of the ganoderma lucidum mycelia polysaccharide is high, the extraction of the ganoderma lucidum mycelia polysaccharide by adopting the ganoderma lucidum mycelia can effectively improve the extraction rate of the polysaccharide, the process steps of later-stage polysaccharide purification are simplified, and the comprehensive utilization effect is good.

The preparation method of the ganoderma lucidum mycelium polysaccharide provided by the invention adopts organic solvent for treatment, and can effectively remove ganoderma lucidumThe esters contained in the ganoderma lucidum mycelia lay the foundation for the efficient extraction of ganoderma lucidum mycelia polysaccharide. Supercritical CO₂The extraction technique is to use supercritical CO₂Dissolving Ganoderma mycelia polysaccharide by supercritical CO under pressure and temperature₂The ganoderma lucidum mycelium polysaccharide is extracted under the supercritical state. The complex enzyme consisting of cellulase, keratinase and hemicellulase is adopted to treat the water extract, so that protein, amino acid and nucleic acid in the water extract can be effectively removed, the purity of ganoderma lucidum mycelium polysaccharide is improved, and the enzyme can be effectively inactivated by heating to 100-120 ℃ after enzymolysis and preserving heat for 1 h. And finally, removing protein and small molecular impurities by adopting an ethanol precipitation method, and further improving the purity of the ganoderma lucidum mycelium polysaccharide.

Compared with the prior art, the invention has the following advantages;

(1) the extraction rate of the ganoderma lucidum mycelium polysaccharide prepared by the preparation method of the ganoderma lucidum mycelium polysaccharide provided by the invention reaches more than 15.4%, and the purity of the ganoderma lucidum mycelium polysaccharide reaches more than 76.1%;

(2) the invention adopts supercritical CO₂The extraction technology and the enzymolysis technology are complementary to each other for polysaccharide extraction of the ganoderma lucidum mycelia, so that the extraction rate of the polysaccharide is improved, the waste of raw materials is avoided, and the production cost is saved;

(3) the preparation method of the ganoderma lucidum mycelium polysaccharide provided by the invention is simple in process, controllable in conditions and easy to realize industrial production; the prepared ganoderma lucidum mycelium polysaccharide is applied to the preparation of health care products and medicines, has good efficacy and obvious economic benefit and concordant benefit.

Detailed Description

The present invention will be further described below by way of specific embodiments, but the present invention is not limited to only the following examples. Various modifications can be made by those skilled in the art based on the basic idea of the invention, but it is within the scope of the invention as long as it does not depart from the basic idea of the invention.