阅读说明：本技术 一种金针菇壳多糖的制备方法 (Preparation method of flammulina velutipes chitin ) 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,具体涉及一种金针菇壳多糖的制备方法。本发明提供的金针菇壳多糖的制备方法采用金针菇为制备原料,通过粉碎、酶解、去除蛋白质、灭酶、提纯、减压浓缩和冷冻干燥等工艺步骤制备金针菇壳多糖。本发明提供的金针菇壳多糖的制备方法提取到的金针菇壳多糖的提取率达到12％,提取到的金针菇壳多糖的纯度达到73.8％。另外,本发明提供的金针菇壳多糖的制备方法简单易操作,条件可控,易于实现工业化生产。(The invention belongs to the technical field of plant extraction, and particularly relates to a preparation method of flammulina velutipes chitin. The preparation method of the flammulina velutipes chitin provided by the invention adopts flammulina velutipes as a preparation raw material, and the flammulina velutipes chitin is prepared by the process steps of crushing, enzymolysis, protein removal, enzyme inactivation, purification, concentration under reduced pressure, freeze drying and the like. The extraction rate of the flammulina velutipes chitin extracted by the preparation method of the flammulina velutipes chitin provided by the invention reaches 12%, and the purity of the extracted flammulina velutipes chitin reaches 73.8%. In addition, the preparation method of the flammulina velutipes chitin provided by the invention is simple and easy to operate, has controllable conditions, and is easy to realize industrial production.)

1. A preparation method of flammulina velutipes chitin is characterized by comprising the following steps:

s1, cutting the cleaned needle mushrooms to the length of 1-2cm, freeze-drying and grinding to 10 microns to obtain needle mushroom powder;

s2, adding 5-8 times of deionized water into the needle mushroom powder prepared in the step S1, soaking for 4-6 hours, heating to 40-60 ℃, adding complex enzyme, and performing enzymolysis for 1-2 hours to obtain an enzymolysis liquid;

s3, adding trichloroacetic acid into the enzymolysis liquid prepared in the step S2, stirring for 2-4 hours under the condition of 400 revolutions per minute until no precipitate is generated, and filtering to prepare filtrate;

s4, heating the filtrate obtained in the step S3 to 90-95 ℃, preserving heat for 0.5-1h, naturally cooling to 25 ℃, performing centrifugal separation, and performing reduced pressure concentration on supernatant to obtain dry extract;

s5, adding an ethanol solution into the dry extract prepared in the step S4 for reflux extraction for 3-5 hours to prepare a mixed solution;

s6, concentrating the mixed solution prepared in the step S5 under reduced pressure, and freeze-drying to obtain the compound.

2. The method for preparing flammulina velutipes chitin according to claim 1, wherein the complex enzyme in step S2 is composed of cellulase, pectinase and decarboxylase in a mass ratio of 2-5:1-3: 3-5.

3. The method for preparing flammulina velutipes chitin according to claim 1, wherein the amount of trichloroacetic acid added in step S3 is 5-10% of the mass of the enzymatic hydrolysate.

4. The preparation method of flammulina velutipes chitin according to claim 1, wherein the volume fraction of the ethanol solution in step S5 is 85%, and the mass ratio of the dry extract to the ethanol solution is 1: 25-35.

5. The method of preparing Flammulina velutipes chitin according to claim 1, wherein the vacuum degree of the vacuum concentration in the steps S4 and S6 is-0.1 to-0.15 MPa, and the temperature is 80 to 90 ℃.

Technical Field

The invention belongs to the technical field of plant extraction, and particularly relates to a preparation method of flammulina velutipes chitin.

Background

The famous flammulina velutipes is called as flammulina velutipes, also called as flammulina velutipes, because the stipe is slender and is similar to that of flammulina velutipes. The needle mushroom contains complete essential amino acid components, particularly abundant lysine and arginine, and high zinc content, and has good effect in enhancing intelligence, especially for children height and intelligence development. The needle mushroom also contains a substance called as needle mushroom essence, which has the effects of enhancing the resistance of the organism to cancer cells, reducing cholesterol, preventing liver diseases and gastrointestinal ulcer, enhancing the body vital qi, preventing diseases and building body after being eaten for a long time. The needle mushroom can effectively enhance the biological activity of organisms, promote metabolism in vivo, is beneficial to the absorption and utilization of various nutrient substances in things, and is also beneficial to growth and development. In addition, the golden mushroom can also inhibit blood fat from rising, reduce cholesterol, and prevent and treat cardiovascular and cerebrovascular diseases, the edible golden mushroom has the effects of resisting fatigue, resisting bacteria and diminishing inflammation, removing heavy metal salt substances and resisting tumors, and the golden mushroom can prevent and treat liver diseases and gastric and intestinal ulcers and is also suitable for being eaten by hypertension patients, obese people and middle-aged and old people.

The flammulina velutipes polysaccharide is a main active ingredient of flammulina velutipes, is a polymer formed by connecting more than 10 monosaccharides through glycosidic bonds, has various functions of good immunoregulation, tumor resistance, liver protection, memory enhancement and the like, and becomes a hotspot in the research fields of food science, natural products, biochemistry and life science. In addition, the flammulina velutipes polysaccharide also has excellent anti-fatigue effect and moisturizing function, and researches show that the flammulina velutipes polysaccharide has better capability of preventing water loss in the horny layer of the skin than glycerin.

The flammulina velutipes chitin is one of flammulina velutipes polysaccharides, exists in cell walls of flammulina velutipes, is a nitrogenous polysaccharide biopolymer, has the effects of bidirectional immune regulation and liver function strengthening, is nutritional and health-care, safe and free of toxic and side effects, is named as the sixth life element after five life elements of sugar, protein, lipid, vitamin and mineral substances are removed by the international medical nutrition food society, and is widely concerned. However, the extraction process of the flammulina velutipes chitin is difficult and heavy because the flammulina velutipes chitin is insoluble in water, dilute acid, alkali solution and organic solvent.

Patent application No. ZL200910045923.8 discloses a preparation method of chitosan in fungal cell walls, which comprises the steps of taking actinocor taiwanensis fungi as raw materials, obtaining bacterial liquid through scratching, shaking culture and fermentation culture, obtaining wet thalli through centrifugal separation, and obtaining the chitosan through separation, drying, crushing, alkali treatment, acid treatment and drying. The preparation method has low energy consumption, easy culture and extraction and simple process, but has low extraction efficiency, and the purity of the extracted chitosan can only reach 50 percent, and the comprehensive utilization effect is poor, thus being not beneficial to industrial production.

In conclusion, the prior art generally has the technical problems of difficult extraction, low extraction efficiency, low product purity, poor comprehensive utilization effect, difficult industrial production and the like.

Disclosure of Invention

In order to overcome the defects of the prior art, the invention aims to provide a preparation method of flammulina velutipes chitin. The preparation method of the flammulina velutipes chitin provided by the invention has the advantages of simple and easy operation process, low extraction efficiency, high product purity, high comprehensive utilization rate, unnecessary waste avoidance, cost saving and easy realization of industrial production.

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

a preparation method of flammulina velutipes chitin specifically comprises the following steps:

s1, cutting the cleaned needle mushrooms to the length of 1-2cm, freeze-drying and grinding to 10 microns to obtain needle mushroom powder;

s2, adding 5-8 times of deionized water into the needle mushroom powder prepared in the step S1, soaking for 4-6 hours, heating to 40-60 ℃, adding complex enzyme, and performing enzymolysis for 1-2 hours to obtain an enzymolysis liquid;

s3, adding trichloroacetic acid into the enzymolysis liquid prepared in the step S2, stirring for 2-4 hours under the condition of 400 revolutions per minute until no precipitate is generated, and filtering to prepare filtrate;

s4, heating the filtrate obtained in the step S3 to 90-95 ℃, preserving heat for 0.5-1h, naturally cooling to 25 ℃, performing centrifugal separation, and performing reduced pressure concentration on supernatant to obtain dry extract;

s5, adding an ethanol solution into the dry extract prepared in the step S4 for reflux extraction for 3-5 hours to prepare a mixed solution;

s6, concentrating the mixed solution prepared in the step S5 under reduced pressure, and freeze-drying to obtain the compound.

Further, the complex enzyme in the step S2 is composed of cellulase, pectinase and decarboxylase according to the mass ratio of 2-5:1-3: 3-5.

Further, the adding amount of the trichloroacetic acid in the step S3 is 5-10% of the mass of the enzymolysis liquid.

Further, the volume fraction of the ethanol solution in the step S5 is 85%, and the mass ratio of the dry extract to the ethanol solution is 1: 25-35.

Further, the vacuum degree of the vacuum concentration in the step S4 and the step S6 is-0.1 to-0.15 Mpa, and the temperature is 80 to 90 ℃.

The preparation method of the flammulina velutipes chitin provided by the invention adopts flammulina velutipes as a preparation raw material, and the flammulina velutipes chitin is prepared by the process steps of crushing, enzymolysis, protein removal, enzyme inactivation, purification, concentration under reduced pressure, freeze drying and the like. The cellulase and the pectinase in the complex enzyme can effectively break the cell wall of the flammulina velutipes, so that chitin in the cell wall of the flammulina velutipes is released, and a solid foundation is laid for extracting the flammulina velutipes chitin; the decarboxylase can remove amino acid contained in the flammulina velutipes, and is favorable for improving the purity of the extracted flammulina velutipes chitin. Trichloroacetic acid can form an insoluble salt with protein under acidic conditions, and the trichloroacetic acid can be used as a protein denaturant to change the conformation of the protein, expose more hydrophobic groups and enable the hydrophobic groups to aggregate and precipitate. Heating the supernatant without protein to 90-95 ℃ and preserving heat for 0.5-1h can effectively inactivate enzyme, thereby being beneficial to improving the purity of the flammulina velutipes chitin. And finally, carrying out ethanol reflux extraction on the solution, and removing monosaccharide, oligosaccharide and other small molecular substances in the solution through ethanol reflux in the process so as to further improve the purity of the flammulina velutipes chitin.

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

(1) the extraction rate of the flammulina velutipes chitin extracted by the preparation method of the flammulina velutipes chitin provided by the invention reaches 12%, and the purity of the extracted flammulina velutipes chitin reaches 73.8%;

(2) according to the preparation method of the flammulina velutipes chitin provided by the invention, the cell wall of the flammulina velutipes is effectively cracked by adopting cellulase and pectinase, and then amino acid and protein contained in the flammulina velutipes are removed by decarboxylase and trichloroacetic acid, so that the purity of the flammulina velutipes chitin is effectively improved;

(3) the preparation method of the flammulina velutipes chitin provided by the invention is simple and easy to operate, controllable in condition, stable in process and easy to realize industrial production.

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.