阅读说明：本技术 一种羊肚菌多糖的提取方法及应用 (Extraction method and application of morchella polysaccharide ) 是由 李峻志 霍文严 祁鹏 刘愚 张黎光 戴璐 乔婷 谢薇梅 李安利 于 2019-11-11 设计创作，主要内容包括：本发明公开了一种羊肚菌多糖的提取方法及应用,首先将羊肚菌粉碎后与复合菌剂混合发酵；其中,复合菌剂包括酵母菌和能够分泌几丁质酶的枯草芽孢杆菌；然后将得到的发酵液与复合酶制剂混合,获得酶解液；其中,复合酶制剂包括几丁质酶与酸性蛋白酶；最后,将酶解液离心、取上清并进行浓缩,加入无水乙醇进行醇析、干燥,得到羊肚菌多糖。本发明的提取工艺简单,工艺条件温和,所得的羊肚菌多糖含量高、生物活性强,在促进外周血白细胞及淋巴细胞的增殖方面具有显著效果。(The invention discloses an extraction method and application of morchella polysaccharide, which comprises the steps of firstly, crushing morchella and then mixing the crushed morchella with a composite microbial inoculum for fermentation; wherein the compound microbial inoculum comprises saccharomycetes and bacillus subtilis capable of secreting chitinase; then mixing the obtained fermentation liquor with a complex enzyme preparation to obtain an enzymolysis liquid; wherein the complex enzyme preparation comprises chitinase and acid protease; and finally, centrifuging the enzymolysis liquid, taking the supernatant, concentrating, adding absolute ethyl alcohol, performing alcohol precipitation and drying to obtain the morchella polysaccharide. The extraction process is simple, the process conditions are mild, the obtained morchella polysaccharide is high in content and strong in biological activity, and the morchella polysaccharide has an obvious effect on promoting the proliferation of peripheral blood leucocytes and lymphocytes.)

1. The extraction method of morchella polysaccharide is characterized by comprising the following steps:

step 1: crushing morchella esculenta, mixing with a composite microbial inoculum, and fermenting; wherein the compound microbial inoculum comprises saccharomycetes and bacillus subtilis capable of secreting chitinase;

step 2: mixing the fermentation liquor obtained in the step 1 with a complex enzyme preparation to obtain an enzymolysis liquid; wherein the complex enzyme preparation comprises chitinase and acid protease;

and step 3: centrifuging the enzymolysis solution, collecting supernatant, concentrating, adding anhydrous ethanol, precipitating with ethanol, and drying to obtain Morchella esculenta polysaccharide.

2. The extraction method of Morchella esculenta polysaccharide as claimed in claim 1, wherein the addition mass of the complex microbial inoculum is 1% -5% of the mass of Morchella esculenta.

3. The method for extracting morchella polysaccharide according to claim 1, wherein the mass ratio of the yeast to the bacillus in the composite microbial agent is 1: 2.

4. the extraction method of Morchella esculenta polysaccharide according to claim 1, wherein the fermentation temperature in step 1 is 29-33 ℃, the pH value is 3.5-4.5, and the fermentation time is 12-16 hours.

5. The extraction method of Morchella esculenta polysaccharide as claimed in claim 1, wherein the mass of the complex enzyme preparation is 1.5-2.0% of the Morchella esculenta.

6. The extraction method of morchella polysaccharide as claimed in claim 1, wherein the mass ratio of chitinase to acid protease in the complex enzyme preparation is 1: 1.

7. the extraction method of Morchella esculenta polysaccharide as claimed in claim 1, wherein the temperature of enzymolysis in step 2 is 36-40 ℃, the pH value is 3.5-4.2, and the action time is 8-12 hours.

8. Use of morchella polysaccharide prepared by the preparation method according to any one of claims 1 to 7 for proliferation of lymphocytes and leukocytes.

Technical Field

The invention belongs to the technical field of polysaccharide extraction, and particularly relates to a method for extracting morchella polysaccharide and application thereof.

Background

Morchella is belonging to Ascomycotina, class of Pediclomycetes, order of Pediculariales, family of Morchellacaceae, genus of Morchella, and is named because it is similar to Morchella. The morchella esculenta can be used as both medicine and food, and the traditional Chinese medicine believes that the morchella esculenta has mild property and sweet taste and has the effects of benefiting intestines and stomach, digesting and assisting food. Modern researches show that the morchella contains abundant polysaccharides, proteins, amino acids, vitamins and various trace elements such as zinc, selenium, iron, germanium, copper and the like. Especially morchella polysaccharide, the modern medical research finds that the morchella polysaccharide has the effects of resisting tumor, resisting bacteria, resisting fatigue, enhancing immunity and the like, and has important development and utilization values in the modern medical science.

At present, the extraction of morchella polysaccharide generally comprises a solvent extraction method, an acid extraction method, an alkali extraction method, an enzyme method and the like. The solvent extraction method has low extraction efficiency and is time-consuming; the acid-base extraction method has complicated procedures, and is easy to destroy glycosidic bonds in the polysaccharide, so that the biological activity of the polysaccharide is reduced; the enzymatic method can act on a limited substrate, limiting its wide application.

Disclosure of Invention

Aiming at the technical problems, the invention provides an extraction method and application of morchella polysaccharide, and aims to solve the problems of low extraction efficiency, time consumption, complicated procedure and easiness in reducing the biological activity of polysaccharide in the existing method.

In order to achieve the purpose, the invention adopts the following technical scheme:

a method for extracting Morchella esculenta polysaccharide comprises:

step 1: crushing morchella esculenta, mixing with a composite microbial inoculum, and fermenting; wherein the compound microbial inoculum comprises saccharomycetes and bacillus subtilis capable of secreting chitinase;

step 2: mixing the fermentation liquor obtained in the step 1 with a complex enzyme preparation to obtain an enzymolysis liquid; wherein the complex enzyme preparation comprises chitinase and acid protease;

and step 3: centrifuging the enzymolysis solution, collecting supernatant, concentrating, adding anhydrous ethanol, precipitating with ethanol, and drying to obtain Morchella esculenta polysaccharide.

Specifically, the adding mass of the compound microbial inoculum is 1-5% of the mass of the morchella esculenta.

Specifically, the mass ratio of the yeast to the bacillus in the composite microbial agent is 1: 2.

specifically, in the step 1, the fermentation temperature is 29-33 ℃, the pH value is 3.5-4.5, and the fermentation time is 12-16 hours.

Specifically, the adding mass of the compound enzyme preparation is 1.5-2.0% of the mass of the morchella esculenta.

Specifically, the mass ratio of chitinase to acid protease in the complex enzyme preparation is 1: 1.

specifically, in the step 2, the temperature in the enzymolysis process is 36-40 ℃, the pH value is 3.5-4.2, and the action time is 8-12 hours.

The invention also discloses application of the morchella polysaccharide prepared by the preparation method in proliferation of lymphocytes and leukocytes.

Compared with the prior art, the invention has the beneficial effects that:

the extraction process is simple, the process conditions are mild, the obtained morchella polysaccharide is high in content and strong in biological activity, and the morchella polysaccharide has an obvious effect on promoting the proliferation of peripheral blood leucocytes and lymphocytes.

Detailed Description

The yeast used in the following examples of the present invention was purchased from Angel Yeast, Inc., and the preservation number of the yeast was CGMCC No. 2.1638. The bacillus is also purchased in the market, and the preservation number of the bacillus is CGMCC No. 9166.

The following embodiments of the present invention are provided, and it should be noted that the present invention is not limited to the following embodiments, and all equivalent changes based on the technical solutions of the present invention are within the protection scope of the present invention.