阅读说明：本技术 一种猴头菇子实体粉固态酶解的方法 (Method for solid-state enzymolysis of hericium erinaceus sporophore powder ) 是由 杨焱 马强 吴迪 张忠 陈万超 李文 张赫男 汪雯翰 于 2020-12-23 设计创作，主要内容包括：本发明提供了一种猴头菇子实体粉固态酶解的方法,属于食用菌深加工领域,本发明将猴头菇子实体粉用缓冲液调糊,添加纤维素酶、β-葡萄糖苷酶和壳聚糖酶组成的复合酶进行酶解,然后烘干灭酶得到猴头菇子实体粉酶解产物。本发明通过酶解处理猴头菇子实体粉,可以较大程度的释放多糖,增加猴头菌产品的附加值,同时可以减少有机试剂的使用,降低成本,减少对环境污染。(The invention provides a solid-state enzymolysis method for hericium erinaceus sporophore powder, belonging to the field of deep processing of edible fungi. According to the invention, the hericium erinaceus sporophore powder is subjected to enzymolysis treatment, so that the polysaccharide can be released to a greater extent, the additional value of hericium erinaceus products is increased, the use of organic reagents can be reduced, the cost is reduced, and the environmental pollution is reduced.)

1. A method for solid enzymolysis of hericium erinaceus sporophore powder is characterized by comprising the following steps: adopting cellulase: β -glucosidase: carrying out enzymolysis on the complex enzyme with the mass ratio of chitosanase of 1-3: 1-3.

2. The method of claim 1, wherein: the cellulase comprises: β -glucosidase: the mass ratio of the chitosanase is 1-2: 2.

3. The method of claim 1, wherein: the addition amount of the complex enzyme is 100-200U/g.

4. The method of claim 1, wherein: the enzymolysis time is 30-70 min.

5. The method according to claim 1, wherein the enzymolysis temperature is 40-60 ℃.

6. The method of claim 1, wherein: the pH value of the enzymolysis is 4-6.

7. The method of claim 1, wherein: and (3) diluting the complex enzyme by using a buffer solution, adding the diluted complex enzyme into the hericium erinaceus sporophore powder, adding the buffer solution into the hericium erinaceus sporophore powder, mixing the hericium erinaceus sporophore powder into paste, and starting enzymolysis.

8. The method of claim 7, wherein: the total addition amount of the buffer solution in each gram of hericium erinaceus sporophore powder is 3-5 mL.

9. The method of claim 1, wherein: drying and inactivating enzyme of the enzymolysis product.

10. The method of claim 9, wherein: the drying and enzyme deactivation conditions are as follows: and inactivating the enzyme in an oven at 75-90 ℃ for 6-10 hours.

Technical Field

The invention belongs to the field of deep processing of edible fungi, and particularly relates to a solid-state enzymolysis method for hericium erinaceus sporophore powder.

Background

Hericium erinaceus (Hericium erinaceus), also known as Hericium erinaceus, Hericium erinaceus and the like, is a large fungus, belongs to the class Basidiomycetes, the order Polyporales, the family Hydnaceae and the genus Hericium, is named because the fruiting body of the Hericium erinaceus is shaped like the head of a monkey, and the nutrient components contained in the Hericium erinaceus are the first and second place compared with other edible fungi artificially cultured at present.

Hericium erinaceus is a famous dish in Chinese banquet and is widely cultivated artificially at present. Hericium erinaceus contains 8 amino acids, polysaccharides and polypeptides which are necessary for human bodies, has the effects of invigorating stomach, improving immunity and the like (Mao Xiaolan [ M ], Chinese mushroom fungus, science publishers, 2009), is a medicine and food dual-purpose fungus, can be used as a raw material of medicines and functional foods, such as a Hericium erinaceus tablet, a Hericium erinaceus oral liquid and the like, and can be used as a food additive for food development, such as a Hericium erinaceus biscuit, a Hericium erinaceus rice paste and the like. However, when hericium erinaceus is developed as a functional food, it is usually necessary to extract it to obtain an extract having a high polysaccharide content; when the product is used as food, the extraction cost is too high, and the product is often a superfine powder product directly utilizing the fruit body of the product. However, the fruiting body of Hericium erinaceus contains a large amount of fibers, cannot be degraded and absorbed by human body, has limited relative content and release of effective components, can achieve the effect only by using a certain amount of fruiting body powder, and increases the burden of human body administration and the limitation of product preparation. However, research and effective technical means are still lacked aiming at how to obtain high-quality hericium erinaceus powder and further develop and utilize the hericium erinaceus powder.

Therefore, how to effectively treat the fruiting body of the hericium erinaceus and improve the release amount of polysaccharide of the hericium erinaceus becomes a technical problem which needs to be solved urgently in the field.

Disclosure of Invention

In view of the above, the present invention aims to provide a method for solid enzymolysis of hericium erinaceus sporophore powder, which can release polysaccharides to a greater extent and increase the added value of hericium erinaceus products.

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

the invention provides a solid enzymolysis method of hericium erinaceus sporophore powder, which comprises the following steps: adopting cellulase: β -glucosidase: carrying out enzymolysis on the complex enzyme with the mass ratio of chitosanase of 1-3: 1-3.

Preferably, the cellulase: β -glucosidase: the mass ratio of the chitosanase is 1-2: 2.

Preferably, the addition amount of the complex enzyme is 100-200U/g.

Preferably, the enzymolysis time is 30-70 min.

Preferably, the enzymolysis temperature is 40-60 ℃.

Preferably, the pH value of the enzymolysis is 4-6.

Preferably, the complex enzyme is diluted by a buffer solution and then added into the hericium erinaceus sporophore powder, and then the buffer solution is added to mix the hericium erinaceus sporophore powder into paste, and then enzymolysis is started.

Preferably, the total addition amount of the buffer solution in each gram of hericium erinaceus sporophore powder is 3-5 mL.

Preferably, the enzymatic hydrolysate is dried and inactivated.

Preferably, the drying enzyme deactivation conditions are as follows: and inactivating the enzyme in an oven at 75-90 ℃ for 6-10 hours.

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

the invention adopts cellulase, beta-glucosidase and chitosanase to mix and use in a proper proportion, cellulose substances in the cell wall and the fruiting body of the hericium erinaceus can be obviously decomposed under proper conditions, the release of polysaccharide effective substances is promoted, the polysaccharide content is increased to 6-8%, the polysaccharide content is increased by more than 120% compared with the traditional process, theoretical basis is provided for the research of the physiological activity of the hericium erinaceus polysaccharide, and the invention has important significance for the development of health care products related to the hericium erinaceus polysaccharide.

The invention utilizes the compound enzymolysis technology to carry out solid degradation on the hericium erinaceus sporophore powder, has the characteristics of high conversion rate, strong specificity, mild action condition, simple operation and the like, so as to obtain more hericium erinaceus polysaccharide release, absorption and utilization, increase the additional value of hericium erinaceus products, and provide a technical method for the development and utilization of the hericium erinaceus sporophore powder food.

The enzyme degradation method adopted by the invention can effectively reduce the use of organic reagents, reduce the production cost, cause no pollution to the environment and have wide industrial application prospect.

Drawings

FIG. 1 shows the effect of different kinds of single enzymes on the release amount of polysaccharide from fruiting body powder of Hericium erinaceus;

FIG. 2 shows the effect of complex enzymes with different proportions on the release amount of polysaccharide from fruiting body powder of Hericium erinaceus;

FIG. 3 shows the effect of different complex enzyme addition amounts on the release amount of polysaccharide from fruiting body powder of Hericium erinaceus;

FIG. 4 shows the influence of different enzymolysis temperatures on the release amount of polysaccharide from fruiting body powder of Hericium erinaceus;

FIG. 5 shows the effect of different enzymolysis times on the release amount of polysaccharide from fruiting body powder of Hericium erinaceus;

FIG. 6 shows the effect of different pH values on the release amount of polysaccharide from fruiting body powder of Hericium erinaceus.

Detailed Description

The invention provides a solid enzymolysis method of hericium erinaceus sporophore powder, which comprises the following steps: adopting cellulase: β -glucosidase: carrying out enzymolysis on the complex enzyme with the mass ratio of chitosanase of 1-3: 1-3.

In the invention, the cellulase (beta-1, 4-glucan-hydrolase) is a general name of a group of enzymes for degrading cellulose to generate glucose, is not a monomer enzyme, is a multi-component enzyme system with a synergistic effect, is a complex enzyme, mainly comprises exo-beta-glucanase, endo-beta-glucanase, beta-glucosidase and the like, accelerates the dissolution rate of water-soluble polysaccharide in the complex enzyme, and improves the release effect of the polysaccharide; the beta-glucosidase can be hydrolyzed and combined with a terminal non-reducing beta-D-glucose bond, and simultaneously releases beta-D-glucose and corresponding aglycone; the chitosan can obviously aim at the fungal cell wall, promote the decomposition of the fungal cell wall and release polysaccharide. The research of the invention finds that the three enzymes are used in a compound way, have the synergistic effect, can obviously decompose cellulose substances in the cell wall and the fruit body of the hericium erinaceus and promote the release of polysaccharide. The specific sources of cellulase, beta-glucosidase and chitosanase are not limited in the present invention.

In the invention, the complex enzyme comprises cellulase: β -glucosidase: the mass ratio of the chitosanase is preferably 1-2: 2, and more preferably 1:1: 1.

The hericium erinaceus fruit body powder can be obtained by pretreating hericium erinaceus fruit bodies. As an optional implementation mode, after unqualified fruiting bodies such as mildewed and deteriorated fruiting bodies and impurities of the hericium erinaceus are removed, the hericium erinaceus is aired and dried until the water content is lower than 8-10%, dried hericium erinaceus fruiting bodies are crushed and sieved by a sieve with 10-12 meshes to prepare hericium erinaceus fruiting body powder, and the hericium erinaceus fruiting body powder is placed in a dry and cool place for later use. The hericium erinaceus fruiting body powder can also be obtained by direct purchase. The specific source of the hericium erinaceus fruiting body/fruiting body powder is not limited in the invention.

The complex enzyme is diluted by buffer solution and then added into the hericium erinaceus sporophore powder, the buffer solution is added for mixing paste, enzymolysis is started, and the total adding amount of the buffer solution in each gram of hericium erinaceus sporophore powder is 3-5 mL, preferably 4 mL. As an optional implementation mode, 2-5 mL of disodium hydrogen phosphate-citric acid buffer solution with the pH value of 4-6 is used for diluting the complex enzyme, the complex enzyme diluent is added into 1-2 g of hericium erinaceus fruit body dry powder, 1-2 mL of the buffer solution is added, the hericium erinaceus fruit body powder and the complex enzyme solution are mixed uniformly to be pasty, and enzymolysis reaction is started. The present invention is not limited to the specific selection of the buffer type.

In the invention, the addition amount of the complex enzyme is 100-200U/g, preferably 150-180U/g, and more preferably 175U/g.

In the invention, the enzymolysis time is 30-70 min, preferably 40-60 min, and more preferably 50 min.

In the invention, the enzymolysis temperature is 40-60 ℃, preferably 45-55 ℃, and more preferably 50 ℃.

In the invention, the pH value of enzymolysis is 4-6, preferably 4.5-5.5, and more preferably 5.0.

The invention dries and inactivates the enzymolysis product. Preferably, the enzymatic hydrolysate is placed in an oven at 75-90 ℃ to inactivate enzyme for 6-10 h, so that the dried and enzyme-inactivated enzymatic hydrolysate is obtained; the temperature of the oven is further preferably 80-84 ℃, and the enzyme deactivation time is further preferably 8 hours.

In the specific implementation process of the invention, the hericium erinaceus sporocarp powder is provided by Shanghai Baixin biotechnology limited; cellulase (500000U/g, C805042, available from Shanghai Michelin Biotechnology Ltd.), beta-glucosidase (50000U/g, S10048, available from Shanghai-derived leaf Biotechnology Ltd.), chitosanase (100000U/g, S25847, available from Shanghai-derived leaf Biotechnology Ltd.).

The technical solution of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. It is to be understood that the described embodiments are merely exemplary of the invention, and not restrictive of the full scope of the invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

Example 1

In the embodiment, a single-factor control experiment is carried out on the addition types of enzymes during solid enzymolysis of the hericium erinaceus sporophore powder:

experimental group 1: cellulase: β -glucosidase: the mass ratio of the chitosanase is 1:1: 1;

experimental group 2: cellulase: β -glucosidase: the mass ratio of the chitosanase is 1:1: 3;

experimental group 3: cellulase: β -glucosidase: the mass ratio of the chitosanase is 1: 2: 2;

experimental group 4: cellulase: β -glucosidase: the mass ratio of the chitosanase is 2: 1: 2;

experimental group 5: cellulase: β -glucosidase: the mass ratio of the chitosanase is 2: 2: 1;

experimental group 6: cellulase: β -glucosidase: the mass ratio of the chitosanase is 2:3: 1;

experimental group 7: cellulase: β -glucosidase: the mass ratio of the chitosanase is 3: 1: 1;

experimental group 8: cellulase: β -glucosidase: the mass ratio of the chitosanase is 3: 2: 1;

experimental group 9: cellulase: β -glucosidase: the mass ratio of the chitosanase is 3: 3: 2;

control group 1: a cellulase;

control group 2: beta-glucosidase;

control group 3: chitosanase;

blank control group: no enzyme was added.

13g of hericium erinaceus sporophore dry powder is randomly divided into 13 parts, and the 13 parts respectively correspond to an experimental group 1-9, a control group 1-3 and a blank control group. Diluting each group of enzyme with 3mL of disodium hydrogen phosphate-citric acid buffer solution with pH5 to obtain enzyme solution (blank control group is only 3mL of buffer solution), correspondingly adding into each group of Hericium erinaceus sporophore dry powder, adding 1mL of disodium hydrogen phosphate-citric acid buffer solution with pH5, mixing to paste, and adjusting pH of the paste to 5. Incubating the reactants in the oven at 50 deg.C for 70min, taking out, and inactivating enzyme in the oven at 80 deg.C for 10 hr.

Drying the dried enzymatic hydrolysate of each group, inactivating enzymes, respectively stirring and uniformly mixing, randomly sampling 100mg of each group, and determining the polysaccharide content.

The enzymolysis process is performed in 3 groups of parallel experiments, the average value of polysaccharide content measured for 3 times is taken, the average polysaccharide content (%) is taken as a survey index, the average polysaccharide content of a control group is detailed in figure 1, and the average polysaccharide content of an experimental group is detailed in figure 2.

As can be seen from fig. 1, the polysaccharide content of the fruit body powder (blank control group) without enzyme treatment is 3.43%, the polysaccharide content of the fruit body powder is 5.63% after solid-state enzymolysis with cellulase, the polysaccharide content of the fruit body powder is 5.32% after solid-state enzymolysis with β -glucosidase, and the polysaccharide content of the fruit body powder is 5.06% after solid-state enzymolysis with chitosanase; the enzymolysis effect of the cellulase is superior to that of beta-glucosidase and chitosanase, and the action effect is obvious compared with that of a group without enzyme treatment, and the enzymolysis effect is improved by 64 percent.

As can be seen from FIG. 2, the results of the enzymolysis with the complex enzymes in different ratios are different, and the polysaccharide contents of the fruiting body powder obtained in experimental groups 1 to 9 are 7.77%, 7.02%, 7.63%, 7.45%, 6.98%, 6.82%, 7.61%, 7.40% and 7.21%, respectively. When the three enzymes are compounded and proportioned in a ratio of 1:1:1, the polysaccharide release is highest and is 7.78 percent; when the three enzymes are compounded and proportioned in a ratio of 2:3:1, the polysaccharide release is the lowest and is 6.82%. Shows that: the complex enzyme treatment is obviously superior to the use of single enzyme, and the polysaccharide content is improved by 21-54%; the polysaccharide content of the hericium erinaceus sporophore powder treated by the compound enzyme is obviously different from that of the hericium erinaceus sporophore powder not treated by the enzyme, and the polysaccharide content is improved by 99-127%.

Note: the polysaccharide content of the hericium erinaceus fruiting body powder in the embodiment is determined by adopting the standard of the Chinese Ministry of agriculture, namely the determination method of edible fungi polysaccharide, and the determination method comprises the following steps:

(1) measuring the content of the hericium erinaceus sporophore total sugar by adopting a phenol-sulfuric acid method, and calculating the extraction rate of the hericium erinaceus sporophore powder total sugar: total sugar content (%) - (C × D × F)/W × 100

In the above formula: c is the glucose concentration (mg/mL) in the test solution, D is the dilution factor of the polysaccharide, F is the conversion factor, and W is the fruiting body mass of Hericium erinaceus.

(2) The content of reducing sugar is determined by a 3, 5-dinitrosalicylic acid method.

(3) Polysaccharide content (%) -total sugar content (%) -reducing sugar content (%)

Example 2

In the embodiment, a single-factor control experiment is carried out on the addition amount of the compound enzyme during solid enzymolysis of the hericium erinaceus sporophore powder:

taking 5g of hericium erinaceus sporophore dry powder, and randomly dividing the hericium erinaceus sporophore dry powder into 5 parts. Diluting each group of enzyme with 3mL of a disodium hydrogen phosphate-citric acid buffer solution with the pH of 5 to obtain enzyme solutions according to 5 levels of the addition of complex enzyme (the mass ratio of cellulase to beta-glucosidase to chitosanase is 1:1:1) of 100U/g, 125U/g, 150U/g, 175U/g and 200U/g to obtain enzyme solutions, correspondingly adding into each group of hericium erinaceus sporophore dry powder, adding 1mL of a disodium hydrogen phosphate-citric acid buffer solution with the pH of 5, uniformly mixing to form paste, and adjusting the pH of the paste to be reacted to 5. Incubating the reactants in the oven at 50 deg.C for 70min, taking out, and inactivating enzyme in the oven at 80 deg.C for 6 hr. Drying the dried enzymatic hydrolysate of each group, inactivating enzymes, respectively stirring and uniformly mixing, randomly sampling 100mg of each group, and determining the polysaccharide content.

3 groups of parallel experiments are carried out in the enzymolysis process, the average value of the polysaccharide content measured for 3 times is taken, the average polysaccharide content (%) is taken as a survey index, and the influence of different adding amounts of the complex enzyme on the polysaccharide release amount of the hericium erinaceus sporocarp is shown in detail in figure 3.

As can be seen from FIG. 3, the polysaccharide release amount of the fruit body powder of Hericium erinaceus increases with the increase of the compound enzyme addition amount, the polysaccharide release amount is the highest at 175U/g, and is 7.49%, and the polysaccharide release amount gradually decreases when the enzyme addition amount exceeds 175U/g. The optimal addition amount of the complex enzyme is 175U/g.

Example 3

In the embodiment, a single-factor control experiment is carried out on the enzymolysis temperature during solid-state enzymolysis of the hericium erinaceus sporophore powder:

taking 5g of hericium erinaceus sporophore dry powder, and randomly dividing the hericium erinaceus sporophore dry powder into 5 parts. Diluting 150U/g of complex enzyme (the mass ratio of cellulase to beta-glucosidase to chitosanase is 1:1:1) with 3mL of disodium hydrogen phosphate-citric acid buffer solution with the pH value of 5 to obtain an enzyme solution, correspondingly adding the enzyme solution into each group of hericium erinaceus sporophore dry powder, respectively adding 1mL of disodium hydrogen phosphate-citric acid buffer solution with the pH value of 5, uniformly mixing to form paste, and adjusting the pH value of the paste to be reacted to 5. Incubating the reactants in oven at 40 deg.C, 45 deg.C, 50 deg.C, 55 deg.C and 60 deg.C for 70min, taking out, and inactivating enzyme in oven at 80 deg.C for 6 hr. Drying the dried enzymatic hydrolysate of each group, inactivating enzymes, respectively stirring and uniformly mixing, randomly sampling 100mg of each group, and determining the polysaccharide content.

The enzymolysis process is carried out in 3 groups of parallel experiments, the average value of the polysaccharide content measured for 3 times is taken, the average polysaccharide content (%) is taken as a survey index, and the influence of different enzymolysis temperatures on the polysaccharide release amount of the hericium erinaceus sporocarp is shown in figure 4.

As can be seen from FIG. 4, the release amount of polysaccharide in the fruit body powder of Hericium erinaceus increases with the increase of the enzymolysis temperature, the release amount of polysaccharide is the highest at 50 ℃ and is 8.11%, and the release amount of polysaccharide gradually decreases when the enzymolysis temperature exceeds 50 ℃. The optimum enzymolysis temperature of the compound enzyme is shown to be 50 ℃.

Example 4

In the embodiment, a single-factor control experiment is carried out on the enzymolysis time during solid-state enzymolysis of the hericium erinaceus sporophore powder:

taking 5g of hericium erinaceus sporophore dry powder, and randomly dividing the hericium erinaceus sporophore dry powder into 5 parts. Diluting 150U/g of complex enzyme (the mass ratio of cellulase to beta-glucosidase to chitosanase is 1:1:1) with 3mL of disodium hydrogen phosphate-citric acid buffer solution with the pH value of 5 to obtain an enzyme solution, correspondingly adding the enzyme solution into each group of hericium erinaceus sporophore dry powder, respectively adding 1mL of disodium hydrogen phosphate-citric acid buffer solution with the pH value of 5, uniformly mixing to form paste, and adjusting the pH value of the paste to be reacted to 5. Respectively incubating the reactants in oven at 50 deg.C for 30min, 50min, 70min, 90min, and 110min, taking out, and placing in oven at 80 deg.C to inactivate enzyme for 6 hr. Drying the dried enzymatic hydrolysate of each group, inactivating enzymes, respectively stirring and uniformly mixing, randomly sampling 100mg of each group, and determining the polysaccharide content.

The enzymolysis process is carried out in 3 groups of parallel experiments, the average value of the polysaccharide content measured for 3 times is taken, the average polysaccharide content (%) is taken as a survey index, and the influence of different enzymolysis time on the polysaccharide release amount of the hericium erinaceus sporocarp is shown in figure 5.

As can be seen from FIG. 5, the release amount of polysaccharide in the fruit body powder of Hericium erinaceus increases with the increase of the enzymolysis time, the release amount of polysaccharide is the highest at 50min, and is 7.32%, and the release amount of polysaccharide gradually decreases with the enzymolysis time exceeding 50 min. The optimum enzymolysis temperature of the compound enzyme is shown to be 50 min.

Example 5

In the embodiment, a single-factor control experiment is carried out on the pH value of the hericium erinaceus sporophore powder during solid enzymolysis:

taking 5g of hericium erinaceus sporophore dry powder, and randomly dividing the hericium erinaceus sporophore dry powder into 5 parts. According to the addition amount of 150U/g of complex enzyme (the mass ratio of cellulase to beta-glucosidase to chitosanase is 1:1:1), diluting with 3mL of disodium hydrogen phosphate-citric acid buffer solution with the pH value of 5 respectively to obtain enzyme solution, correspondingly adding into each group of hericium erinaceus sporophore dry powder, then adding disodium hydrogen phosphate-citric acid buffer solution with the pH value of 1mL 5 respectively, and uniformly mixing to obtain paste. Adjusting the pH value of each group of reactants to be reacted to 4, 4.5, 5, 5.5 and 6 respectively, incubating each group of reactants in an oven at 50 ℃ for 70min, taking out, and placing in an oven at 80 ℃ to inactivate enzyme for 6 h. Drying the dried enzymatic hydrolysate of each group, inactivating enzymes, respectively stirring and uniformly mixing, randomly sampling 100mg of each group, and determining the polysaccharide content.

The enzymolysis process is carried out in 3 groups of parallel experiments, the average value of polysaccharide content measured for 3 times is taken, the average polysaccharide content (%) is taken as a survey index, and the influence of different enzymolysis pH values on the polysaccharide release amount of the hericium erinaceus sporocarp is shown in figure 6.

As can be seen from fig. 6, the release amount of polysaccharide in the fruit body powder of hericium erinaceus increases with the increase of the pH value of the enzymolysis, the release amount of polysaccharide is the highest at pH5, which is 8.76%, and the release amount of polysaccharide gradually decreases after the pH value of the enzymolysis exceeds 5. The pH value of the compound enzyme is shown to be 5.

The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.