阅读说明：本技术 一种食用菌酶解液及其制备方法 (Edible fungus enzymatic hydrolysate and preparation method thereof ) 是由 贾凤娟 王文亮 弓志青 宋莎莎 张剑 于 2020-09-08 设计创作，主要内容包括：本发明属于食用菌提取物技术领域,具体涉及一种食用菌酶解液及其制备方法。本发明所述食用菌酶解液,以黑木耳为原料经纤维素酶解及水提制得,选用的所述黑木耳利用选定培养基质进行培养,并通过添加活性污泥颗粒的促进作用,有效促进了栽培基质中营养成分的吸收和目标物质的形成,不仅大幅提升了黑木耳的产量,更为有效提高了黑木耳中黑木耳多糖的含量,同时有效保证所得黑木耳产品的品质和性能。(The invention belongs to the technical field of edible fungus extracts, and particularly relates to an edible fungus enzymatic hydrolysate and a preparation method thereof. The edible fungus enzymatic hydrolysate is prepared by taking black fungus as a raw material and performing cellulose hydrolysis and water extraction on the black fungus, the selected black fungus is cultured by utilizing a selected culture medium, and the absorption of nutrient components and the formation of target substances in the culture medium are effectively promoted by adding the promoting effect of activated sludge particles, so that the yield of the black fungus is greatly increased, the content of black fungus polysaccharide in the black fungus is more effectively increased, and the quality and the performance of the obtained black fungus product are effectively ensured.)

1. The preparation method of the edible fungus enzymatic hydrolysate is characterized by comprising the steps of cultivating black fungus by using artificial bagged materials and carrying out enzymatic extraction on the obtained black fungus, and specifically comprises the following steps:

(1) taking 50-60 parts by weight of sawdust, 10-20 parts by weight of bran, 5-12 parts by weight of soybean protein isolate, 8-20 parts by weight of brewer's grain alkaline hydrolysate, 5-15 parts by weight of rice residue, 0.5-2 parts by weight of lime, 0.5-2 parts by weight of gypsum and 0.3-0.8 part by weight of propolis, uniformly mixing to obtain the required culture medium, sterilizing at high temperature, and naturally cooling to room temperature for later use;

(2) under an aseptic environment, adding 5-20 parts by weight of activated sludge particles into the sterilized culture medium, uniformly mixing, adding water, fully and uniformly mixing, filling into a cylindrical plastic layer, and tying and sealing two ends of the plastic layer to obtain a required fungus bag for later use;

(3) implanting bacterium liquid containing black fungus strains into the fungus bag by using a quantitative injector to prepare a fungus bag, uniformly puncturing the fungus bag, and placing the obtained fungus bag in a culture room for culture; placing the cultured fungus bags in a hanging bag type greenhouse for cultivation and planting, and picking up the black fungus when the black fungus is mature;

(4) removing surface impurities of picked black fungus, drying to constant weight, adding water, pulping for later use;

(5) adding cellulase into the black fungus slurry for enzymolysis, performing solid-liquid separation after enzyme deactivation, and collecting enzymolysis liquid to obtain the black fungus slurry.

2. The method for preparing the edible fungus enzymatic hydrolysate of claim 1, wherein in the step (1), the brewer's grain alkaline hydrolysate is a product of alkaline hydrolysis of brewer's grain by NaOH solution at 60-80 ℃.

3. The method for preparing edible fungus enzymatic hydrolysate of claim 1 or 2, wherein in the step (1), the rice residue is leftovers of a process for preparing starch sugar from rice.

4. The method for preparing an enzymatic hydrolysate of edible fungi according to any one of claims 1 to 3, wherein in the step (2), the activated sludge granules are aerobic activated sludge granules.

5. The method for preparing an enzymatic hydrolysate of edible fungi according to any one of claims 1 to 4, wherein in the step (2), the mass ratio of the culture substrate to the water is controlled to be 40 to 50%: 50 to 60 percent.

6. The method for preparing an enzymatic hydrolysate of edible fungi according to any one of claims 1 to 5, wherein in the step (3), the amount of the fungus liquid containing Auricularia auricular strains implanted is 5 to 10 wt% of the culture medium.

7. The method for preparing edible fungus enzymatic hydrolysate of any one of claims 1 to 6, wherein in the step (3), the temperature of the culture chamber is controlled to be 25 +/-2 degrees, the humidity is controlled to be 20-30 percent, and the culture chamber is protected from light while a ventilation system is adopted to control carbon dioxide in the culture chamber to be not more than 3000 ppm.

8. The method for preparing an enzymatic hydrolysate of edible fungi according to any one of claims 1 to 7, wherein in the step (4), the pulping step is to add water with an amount of 8 to 10 times the mass of the black fungus for pulping.

9. The method for preparing an enzymatic hydrolysate of edible fungi according to any one of claims 1 to 8, wherein in the step (5), the cellulase is added in an amount of 0.3 to 0.8 wt% based on the mass of the black fungus.

10. An enzymatic hydrolysate of edible fungi prepared by the method of any one of claims 1 to 9.

Technical Field

The invention belongs to the technical field of edible fungus extracts, and particularly relates to an edible fungus enzymatic hydrolysate and a preparation method thereof.

Background

The edible fungus refers to edible mushroom (large-scale fungus) with large fruiting body, and is commonly called mushroom. More than 350 kinds of edible fungi are known in China, wherein more of the edible fungi belong to basidiomycotina, and common edible fungi include shiitake mushroom, straw mushroom, black fungus, tremella, hericium erinaceus, dictyophora phalloidea, tricholoma matsutake, russula, ganoderma lucidum, cordyceps sinensis, truffle, pleurotus nebrodensis, boletus and the like; the minority of the strains belong to ascomycetous subgenus, and the minority mainly comprises morchella, saddle fungus, truffle and the like. The fungi grow in different regions and different ecological environments respectively.

The edible fungi contain various components such as abundant carbohydrates, proteins, various amino acids, steroids and the like, and have various biological activities such as anti-tumor, anti-oxidation, antibiosis, liver protection and the like; meanwhile, the edible fungi are delicious in taste, the by-products of the edible fungi are rich in dietary fibers such as beta-glucan, chitin, hemicellulose, mannose and the like, and have rich nutrition and higher medicinal value, so that the edible fungi become food materials which are widely concerned in recent years, are popular among people and become common food materials on daily dining tables.

The black fungus is a kind of edible fungus with rich nutrition, and is named because it grows on rotten wood and is shaped like human ears. Fresh black fungus is in a colloid sheet shape, is elastic and semitransparent, grows laterally on trees, has the diameter of 5-10 cm, is smooth and concave on the abdominal surface, is slightly curled at the edge, is convex at the back, has superfine villi, and is black brown or dark brown. Shrinking into cutin shape after drying, hard and brittle, dark gray or grey white back; it swells in water and can recover to original shape, and is soft and translucent, and the surface is adhered with smooth mucus. The black fungus is rich in beneficial elements such as iron, calcium, phosphorus, vitamin B1 and the like, has soft texture, delicious taste and rich nutrition, can be eaten as vegetable or meat, not only adds wind for dishes, but also can nourish blood, preserve youthful looks, remove diseases and prolong life. Black fungus is praised as 'meat in vegetables' by modern nutriologists, has a nutritional value comparable with that of food, and is a traditional health food and an export commodity in China.

The polysaccharide component in the black fungus can improve the immunity of a human body, enhance the disease resistance of the human body and protect the damage of human tissue cells according to research; the auricularia auricula polysaccharide can also reduce the blood fat content of an experimental white mouse, and has an antithrombotic effect on experimental rabbits and guinea pigs; the auricularia auricula polysaccharide can reduce the formation of lipochrome, maintain the normal function of human cells and delay aging by reducing the content of cholesterol in blood vessels; the Auricularia polysaccharide also has effects of resisting tumor cell activity and mutation, promoting ulcer healing, and eliminating black spot; in addition, Auricularia auricula polysaccharide also has effects of enhancing superoxide dismutase activity, and can scavenge free radicals in human body. Therefore, how to cultivate and obtain the black fungus with high polysaccharide content and further obtain high-performance extract liquid for downstream product processing has positive significance for the development of edible fungi.

Disclosure of Invention

Therefore, the technical problem to be solved by the invention is to provide the edible fungus enzymatic hydrolysate which has the advantages of high polysaccharide content and better health care performance;

the second technical problem to be solved by the present invention is to provide a method for preparing the above edible fungus enzymatic hydrolysate.

In order to solve the technical problems, the preparation method of the edible fungus enzymatic hydrolysate comprises the steps of cultivating black fungus by using artificial bagged materials and carrying out enzymatic extraction on the obtained black fungus, and specifically comprises the following steps:

(1) taking 50-60 parts by weight of sawdust, 10-20 parts by weight of bran, 5-12 parts by weight of soybean protein isolate, 8-20 parts by weight of brewer's grain alkaline hydrolysate, 5-15 parts by weight of rice residue, 0.5-2 parts by weight of lime, 0.5-2 parts by weight of gypsum and 0.3-0.8 part by weight of propolis, uniformly mixing to obtain the required culture medium, sterilizing at high temperature, and naturally cooling to room temperature for later use;

(2) under an aseptic environment, adding 5-20 parts by weight of active sludge particles into the sterilized culture medium, uniformly mixing, adding water, fully and uniformly mixing, filling into a cylindrical plastic layer, and sealing two ends of the plastic layer by pricking to obtain a required fungus bag for later use;

(3) implanting bacterium liquid containing black fungus strains into the bacterium bag by using a quantitative injector to prepare a bacterium bag, uniformly puncturing the bacterium bag, and placing the obtained bacterium bag in a culture room for culture; placing the cultured fungus bags in a hanging bag type greenhouse for cultivation and planting, and picking up the black fungus when the black fungus is mature;

(4) removing surface impurities of picked black fungus, drying to constant weight, adding water, pulping for later use;

(5) adding cellulase into the black fungus slurry for enzymolysis, performing solid-liquid separation after enzyme deactivation, and collecting enzymolysis liquid to obtain the black fungus slurry.

Specifically, in the step (1), the beer lees alkaline hydrolysate is a product obtained by carrying out alkaline hydrolysis on beer lees at 60-80 ℃ by using a NaOH solution.

Specifically, in the step (1), the rice residue is leftovers of a process for preparing starch sugar from rice.

Specifically, in the step (2), the activated sludge particles are aerobic activated sludge particles.

Specifically, in the step (2), the mass ratio of the culture medium to water is controlled to be 40-50%: 50 to 60 percent.

Specifically, in the step (3), the implantation amount of the fungus liquid containing the black fungus strains accounts for 5-10 wt% of the weight of the culture medium.

Specifically, in the step (3), the temperature of the culture chamber is controlled to be 25 +/-2 degrees, the humidity is controlled to be 20-30 percent, the culture chamber is protected from light, and meanwhile, a ventilation system is adopted to control carbon dioxide in the culture chamber to be not more than 3000 ppm.

Specifically, in the step (4), the pulping step is to add water with the weight of 8-10 times of that of the black fungus for pulping.

Specifically, in the step (5), the addition amount of the cellulase accounts for 0.3-0.8 wt% of the weight of the black fungus.

Specifically, in the step (5), the temperature of the enzymolysis step is 45-50 ℃, and the enzymolysis time is controlled for 2-3 h.

The invention also discloses the edible fungus enzymatic hydrolysate prepared by the method.

The edible fungus enzymatic hydrolysate is prepared by taking black fungus as a raw material and performing cellulose enzymolysis and water extraction, the selected black fungus is cultured by utilizing a selected culture medium, and the promotion effect of activated sludge particles is added, so that the nutrient components of the culture medium are fully utilized. In the culture medium, alkaline hydrolysate obtained by alkaline hydrolysis of beer lees is added, so that the culture medium is rich in nutrition, and the alkaline hydrolyzed components are more suitable for cultivating the edible fungus, thereby effectively improving the yield of the edible fungus and the content of edible fungus polysaccharide in the edible fungus. In the process of cultivating the black fungus, the activated sludge particles are further added, so that the absorption of nutrient components in the culture medium and the formation of target substances are effectively promoted, the yield of the black fungus is greatly increased, the content of black fungus polysaccharide in the black fungus is effectively increased, and the quality and the performance of the obtained black fungus product are effectively ensured.

Detailed Description

The following examples of the invention:

the beer lees alkaline hydrolysis product is prepared by adding beer lees into 5 times of 30 wt% NaOH solution, mixing uniformly, carrying out alkaline hydrolysis at 70 ℃ for 2h, washing the obtained black brown paste to remove alkali liquor, and drying;

the rice residue is leftovers produced in the process of preparing starch sugar from rice, wherein the dry basis of protein is more than or equal to 60 percent, the water content is less than or equal to 10 percent, the total sugar is less than or equal to 20 percent, the fat is less than or equal to 12 percent, and the ash content is less than or equal to 5 percent.

Example 1

The preparation method of the black fungus enzymolysis liquid comprises the following steps:

(1) taking 50 parts by weight of sawdust, 20 parts by weight of bran, 5 parts by weight of soybean protein isolate, 20 parts by weight of beer vinasse alkaline hydrolysate, 5 parts by weight of rice residue, 2 parts by weight of lime, 0.5 part by weight of gypsum and 0.8 part by weight of propolis, uniformly mixing to obtain a required culture medium, sterilizing at high temperature, and naturally cooling to room temperature for later use;

(2) and (2) under an aseptic environment, adding 5 parts by weight of aerobic activated sludge particles into the sterilized culture substrate, uniformly mixing, and mixing according to the weight percentage of 50%: adding water into the culture medium in a mass ratio of 50 wt%, fully and uniformly mixing, filling the mixture into a cylindrical plastic layer (the diameter is 15cm, the length is 25cm), and sealing the two ends of the plastic layer to obtain the required fungus bag for later use;

(3) injecting a bacterium solution (a commercial product) containing black fungus strains accounting for 10 wt% of the weight of the culture medium into the bacterium bag by using a quantitative injector to prepare a bacterium bag, uniformly puncturing the bacterium bag by using 6mm iron nails, and placing the obtained bacterium bag in a culture room for fungus-promoting culture; in the culture room, the temperature is controlled to be 25 +/-2 degrees, the humidity is controlled to be 20-30 percent, the culture room is ventilated in a dark place, and meanwhile, a ventilation system is adopted to control the carbon dioxide in the culture room to be not more than 3000 ppm;

placing the fungus bags with cultured ears in a hanging bag type greenhouse for cultivation: the cultured fungus bags are placed in a hanging bag type greenhouse through binding or vertical suspension, the outer layer of the greenhouse is covered by a plastic film, and meanwhile, a sunshade net is covered on the outer layer of the plastic film to control the temperature in the greenhouse; controlling the humidity of black fungus in the greenhouse to be about 70 +/-5% in the early stage of planting and 85-90% in the later stage by using a spraying facility at the top end inside the greenhouse, and picking the black fungus when the black fungus is mature;

(4) cleaning the picked black fungus with clear water, naturally airing, adding 100g of the treated black fungus into water which accounts for 10 times of the weight of the black fungus, and pulping to obtain black fungus slurry;

(5) adding cellulase accounting for 0.5 wt% of the solid mass of the black fungus into the black fungus slurry, carrying out enzymolysis treatment at 45 ℃ for 2h, heating to 90-100 ℃ after the enzymolysis is finished, and keeping for 20-30min to inactivate enzyme; transferring the enzyme-inactivated Auricularia solid-liquid mixture to a centrifuge, performing solid-liquid separation, and collecting the obtained Auricularia enzymolysis solution.

Example 2

The preparation method of the black fungus enzymolysis liquid comprises the following steps:

(1) taking 60 parts by weight of sawdust, 10 parts by weight of bran, 12 parts by weight of soybean protein isolate, 8 parts by weight of brewer's grain alkaline hydrolysate, 15 parts by weight of rice residue, 0.5 part by weight of lime, 2 parts by weight of gypsum and 0.3 part by weight of propolis, uniformly mixing to obtain a required culture medium, sterilizing at high temperature, and naturally cooling to room temperature for later use;

(2) under the aseptic environment, adding 20 parts by weight of aerobic activated sludge particles into the sterilized culture medium, uniformly mixing, and mixing according to the weight percentage of 50%: taking the culture medium according to the mass ratio of 50 wt%, adding water, fully and uniformly mixing, filling the mixture into a cylindrical plastic layer (the diameter is 15cm, the length is 25cm), and tying and sealing two ends of the plastic layer to obtain the required fungus bag for later use;

(3) injecting a bacterium solution (a commercial product) containing black fungus strains accounting for 10 wt% of the weight of the culture medium into the bacterium bag by using a quantitative injector to prepare a bacterium bag, uniformly puncturing the bacterium bag by using 6mm iron nails, and placing the obtained bacterium bag in a culture room for fungus-promoting culture; in the culture room, the temperature is controlled to be 25 +/-2 degrees, the humidity is controlled to be 20-30 percent, the culture room is ventilated in a dark place, and meanwhile, a ventilation system is adopted to control the carbon dioxide in the culture room to be not more than 3000 ppm;

placing the fungus bags with cultured ears in a hanging bag type greenhouse for cultivation: the cultured fungus bags are placed in a hanging bag type greenhouse through binding or vertical suspension, the outer layer of the greenhouse is covered by a plastic film, and meanwhile, a sunshade net is covered on the outer layer of the plastic film to control the temperature in the greenhouse; controlling the humidity of black fungus in the greenhouse to be about 70 +/-5% in the early stage of planting and 85-90% in the later stage by using a spraying facility at the top end inside the greenhouse, and picking the black fungus when the black fungus is mature;

(4) cleaning the picked black fungus with clear water, naturally airing, adding 100g of the treated black fungus into water which accounts for 10 times of the weight of the black fungus, and pulping to obtain black fungus slurry;

(5) adding cellulase accounting for 0.5 wt% of the solid mass of the black fungus into the black fungus slurry, carrying out enzymolysis treatment at 45 ℃ for 2h, heating to 90-100 ℃ after the enzymolysis is finished, and keeping for 20-30min to inactivate enzyme; transferring the enzyme-inactivated Auricularia solid-liquid mixture to a centrifuge, performing solid-liquid separation, and collecting the obtained Auricularia enzymolysis solution.

Example 3

The preparation method of the black fungus enzymolysis liquid comprises the following steps:

(1) taking 55 parts by weight of sawdust, 15 parts by weight of bran, 8 parts by weight of soybean protein isolate, 15 parts by weight of beer vinasse alkaline hydrolysate, 10 parts by weight of rice residue, 1 part by weight of lime, 1 part by weight of gypsum and 0.5 part by weight of propolis, uniformly mixing to obtain a required culture medium, sterilizing at high temperature, and naturally cooling to room temperature for later use;

(2) under the aseptic environment, adding 12 parts by weight of aerobic activated sludge particles into the sterilized culture medium, uniformly mixing, and mixing according to the weight percentage of 50%: taking the culture medium according to the mass ratio of 50 wt%, adding water, fully and uniformly mixing, filling the mixture into a cylindrical plastic layer (the diameter is 15cm, the length is 25cm), and tying and sealing two ends of the plastic layer to obtain the required fungus bag for later use;

(3) injecting a bacterium solution (a commercial product) containing black fungus strains accounting for 10 wt% of the weight of the culture medium into the bacterium bag by using a quantitative injector to prepare a bacterium bag, uniformly puncturing the bacterium bag by using 6mm iron nails, and placing the obtained bacterium bag in a culture room for fungus-promoting culture; in the culture room, the temperature is controlled to be 25 +/-2 degrees, the humidity is controlled to be 20-30 percent, the culture room is ventilated in a dark place, and meanwhile, a ventilation system is adopted to control the carbon dioxide in the culture room to be not more than 3000 ppm;

placing the fungus bags with cultured ears in a hanging bag type greenhouse for cultivation: the cultured fungus bags are placed in a hanging bag type greenhouse through binding or vertical suspension, the outer layer of the greenhouse is covered by a plastic film, and meanwhile, a sunshade net is covered on the outer layer of the plastic film to control the temperature in the greenhouse; controlling the humidity of black fungus in the greenhouse to be about 70 +/-5% in the early stage of planting and 85-90% in the later stage by using a spraying facility at the top end inside the greenhouse, and picking the black fungus when the black fungus is mature;

(4) cleaning the picked black fungus with clear water, naturally airing, adding 100g of the treated black fungus into water which accounts for 10 times of the weight of the black fungus, and pulping to obtain black fungus slurry;

(5) adding cellulase accounting for 0.5 wt% of the solid mass of the black fungus into the black fungus slurry, carrying out enzymolysis treatment at 45 ℃ for 2h, heating to 90-100 ℃ after the enzymolysis is finished, and keeping for 20-30min to inactivate enzyme; transferring the enzyme-inactivated Auricularia solid-liquid mixture to a centrifuge, performing solid-liquid separation, and collecting the obtained Auricularia enzymolysis solution.

Comparative example 1

The preparation method of the black fungus enzymatic hydrolysate in the comparative example is the same as that in example 3, the difference is that the active sludge particles are not added in the black fungus cultivation, and the EM bacterial liquid is inoculated when the black fungus liquid is inoculated; the EM bacterial liquid is obtained by inoculating EM original liquid into a proper culture medium (5% of glucose, 2% of peptone, 0.2% of glutamine, 0.2% of water-soluble vitamin and adjusting pH to 7.0) and fermenting at normal temperature.

Comparative example 2

The preparation method of the black fungus enzymatic hydrolysate in the comparative example is the same as that in example 3, and only differs from the preparation method in that carboxymethyl cellulose is used for replacing the beer lees alkaline hydrolysate.

Comparative example 3

The preparation method of the black fungus enzymatic hydrolysate of the comparative example is the same as that of example 3, and only differs from that of example 3 in that beer grains are used for replacing the beer grain alkaline hydrolysate.

Examples of the experiments

1. Comparison of Auricularia auricula-judae Performance

The product morphology detection of the black fungus cultivated in the above examples 1-3 and comparative examples 1-3 is performed, and the results are shown in the following table 1.

TABLE 1 Auricularia auricula growth and product Performance test results

The data in the table above show that the black fungus culture medium provided by the invention makes full use of the nutritional ingredients of the culture medium by selecting the culture medium and adding the promotion effect of the activated sludge particles, so that the yield of the black fungus is greatly increased, and the quality and performance of the obtained black fungus product are effectively ensured.

2. Auricularia auricula polysaccharide content

The black fungus enzymatic hydrolysate prepared in the above examples 1-3 and comparative examples 1-3 were collected, and the content of black fungus polysaccharide contained in the enzymatic hydrolysate was determined (i.e. the polysaccharide content extracted from each 100g of black fungus), and the test results are shown in table 2 below.

The specific determination method refers to a phenol-sulfuric acid colorimetric method disclosed in Chinese patent CN108641011A for determining the content of Auricularia auricular polysaccharide.

TABLE 2 polysaccharide content in the enzymatic hydrolysate

Numbering	Polysaccharide content/g of enzymolysis liquid
		Example 1	7.68
Example 2	7.79
		Example 3	7.84
Comparative example 1	7.13
		Comparative example 2	6.03
Comparative example 3	6.59

As can be seen from the data in the table, the black fungus culture medium is selected, and the promotion effect of adding activated sludge particles is utilized, so that the nutrient components of the culture medium are fully utilized, and the content of black fungus polysaccharide in the black fungus is effectively improved.

It should be understood that the above-described embodiments are merely examples for clarity of description and are not intended to limit the scope of the embodiments. Other variations and modifications will be apparent to persons skilled in the art in light of the above description. This list is neither intended to be exhaustive nor exhaustive. And obvious variations or modifications therefrom are within the scope of the invention.