A preparation process and device for edible mushroom small molecule peptide

Abstract

The present invention discloses a preparation process of edible mushroom small molecule peptides and its supporting device, wherein the mushroom small molecule peptides are prepared by a high voltage pulse electric field assisted composite enzyme method, the extraction efficiency is high, and the peptides are suitable for large-scale industrial production. The present invention also discloses the edible mushroom small molecule peptides prepared by the process and their application.

Description

Technical Field

[0001] The present application belongs to the field of preparation of small molecule peptides, and specifically relates to a preparation process and device of small molecule peptides of edible mushrooms. Background Art

[0002] In recent years, proteins from edible fungi have gradually attracted the attention of the scientific community and the food industry. Compared with proteins from other sources, edible mushroom proteins have obvious advantages: due to the wide variety of edible fungi, their protein types are also very rich; edible fungi proteins have mild, low-sensitivity, and antioxidant effects; edible fungi proteins have better thermal stability and pH stability, and can be used as raw materials for industrial production; the cultivation environment of edible fungi is environmentally friendly and the cultivation speed is fast, etc. On this basis, edible mushroom peptides are also favored by consumers because of their smaller molecular weight, which can be more directly absorbed and exerted by the human body. As a powerful substitute for traditional animal and plant proteins, edible fungi proteins and edible fungi peptides have been widely used in food processing, biomedicine, and chemical industries.

[0003] The prior art provides a variety of conventional methods for extracting proteins and peptides from edible mushrooms, such as solvent extraction, enzyme extraction, and ultrasonic-assisted extraction. The solvent extraction method usually selects the corresponding acid, alkali, salt, and organic solvent to extract protein according to the substrate, but the protein extraction rate mainly depends on the pH value and salt concentration, and the extraction environment required by different substrates varies greatly. The enzyme extraction method releases proteins from cells in a soluble form through the action of enzymes, but the enzyme extraction rate is greatly affected by the type, proportion, temperature, and time of the enzyme. The ultrasonic-assisted extraction method promotes hydration through ultrasound, causing an increase in cell wall pores to improve protein extraction efficiency, but the actual extraction effect is also limited by the type of ultrasonic reactor and the ultrasonic frequency and power.

[0004] There are also documents that disclose the combination of multiple methods to extract peptides, such as the use of high-voltage pulsed electric field-assisted enzyme extraction. The process steps and parameters of such methods are not adjusted and optimized according to the substrate and cannot be simply transplanted to the extraction of edible mushroom peptides.

[0005] Therefore, there is an urgent need in the art to develop a process for preparing edible mushroom peptides with high extraction efficiency and industrial production.

[0006] The present invention also has the advantage of providing a device for preparing edible mushroom peptides, which can realize efficient and large-scale extraction of edible mushroom peptides.

[0007] In addition, another advantage of the present invention is to provide a mushroom small molecule peptide, which has a higher peptide content and optimized peptide molecule distribution. Summary of the invention

[0008] Brief description of the invention

[0009] In one aspect, the present invention provides a process for preparing a small molecule peptide of edible mushrooms, comprising:

[0010] a) Pretreatment: Take dried edible mushrooms, crush them by ultra-fine wall breaking to obtain ultra-fine mushroom powder, add distilled water in a mixing tank at a solid-liquid ratio of 1:10-20, stir and dissolve to obtain mushroom solution;

[0011] b) High-voltage pulse electric field treatment: The mushroom solution in the preparation tank is sent to the pulse electric field treatment chamber through a peristaltic pump for high-voltage pulse electric field treatment, with the electric field intensity controlled at 10-20 kV / cm and the number of pulses at 6-14;

[0012] c) Protein extraction: add sodium hydroxide solution to the mushroom liquid treated with high-voltage pulse electric field to adjust the pH to 8.0, let it stand for 5-15 minutes, then add hydrochloric acid solution to adjust the pH to 4.5, centrifuge for 5-15 minutes and collect the precipitate;

[0013] d) compound enzyme hydrolysis: placing the precipitate in an enzymolysis tank, adding distilled water to prepare a 1% to 5% solution, adjusting the pH to 8.0, heating to 40±0.5° C., and adding 3000-7000 U / g of compound enzyme for 1 to 3 hours of enzymolysis, wherein the compound enzyme comprises at least two proteases selected from the group consisting of papain, alkaline protease, neutral protease, pepsin, trypsin, cathepsin and subtilisin;

[0014] e) collecting the supernatant: heating the enzymatically hydrolyzed liquid to 85-95° C. and maintaining the temperature for 5-15 minutes, cooling the liquid and centrifuging at 4000-6000 r / min for 10-20 minutes to collect the supernatant; and

[0015] f) Filtration: Filter the supernatant through a filter membrane with a molecular weight ≥ 3KD to obtain a mushroom small molecule peptide concentrate.

[0016] Preferably, in the process for preparing small molecule peptides of edible mushrooms described in the present invention, in the step d), the composite enzyme comprises papain, alkaline protease and neutral protease, and the ratio of the three proteases in the composite enzyme is 1:(1.5-2):(1-1.5).

[0017] Preferably, in the process for preparing the edible mushroom small molecule peptide of the present invention, the edible mushroom is a Pleurotus genus fungus, more preferably Pleurotus eryngii.

[0018] In another aspect, the present invention provides a device for preparing small molecule peptides of edible mushrooms, comprising:

[0019] A mixing tank is used to contain mushroom powder and distilled water to dissolve and obtain a mushroom solution;

[0020] a peristaltic pump, the inlet of which is connected to the mixing tank, and is used to deliver the mushroom solution from the mixing tank to the electric pulse generator;

[0021] The electric pulse generator is connected to the outlet of the peristaltic pump and is used to perform high-voltage pulse electric field treatment on the mushroom solution;

[0022] A centrifugal device for centrifuging the solution after protein extraction; and

[0023] Enzyme hydrolysis tank, used for compound enzyme hydrolysis reaction.

[0024] On the other hand, the present invention provides a mushroom small molecule peptide prepared by the aforementioned edible mushroom small molecule peptide preparation process, wherein the mushroom small molecule peptide is a concentrated liquid or a dry powder.

[0025] In another aspect, the present invention provides a cosmetic comprising the aforementioned mushroom small molecule peptide. DETAILED DESCRIPTION OF THE INVENTION

[0027] The present invention provides a specific small molecule peptide preparation process for edible mushrooms, which uses a high-voltage pulse electric field plus a specific composite enzyme for hydrolysis, and is equipped with corresponding process parameters and steps, and is suitable for efficient and large-scale extraction of edible mushroom small molecule peptides. Without being bound by theory, the applicant was surprised to find that in the preparation process of the present invention, the high-voltage pulse electric field breaks through the cell membrane of edible mushrooms, causing the cell contents to seep out, and the specific high-voltage pulse electric field intensity can accelerate the dissolution of soluble proteins in the cells, thereby improving the productivity of the peptides; at the same time, in the selection of enzymes, a composite enzyme of a specific component is used for hydrolysis reaction, which ultimately increases the peptide content of the product and optimizes its peptide molecular weight distribution, so that it is easy for the human body to absorb.

[0028] It is worth mentioning that the composite enzyme hydrolysis method adopted in the present invention has obvious advantages over traditional single enzyme method and multi-step enzyme method. Compared with the single enzyme method, the composite enzyme method forms polypeptides for multiple restriction sites at the same time, with high production efficiency and small molecular weight of peptides. Especially in the case of edible mushrooms as substrates, because of its abundant protein species, it is necessary to configure specific composite enzymes, thereby improving the productivity and product quality of peptides. Compared with the multi-step enzyme method, the composite enzyme method has a shorter overall reaction time, simplified steps, simple operation, no need to adjust pH repeatedly, and is therefore more suitable for efficient, large-scale industrial production.

[0029] Preparation process

[0030] The process for preparing edible mushroom small molecule peptides of the present invention comprises:

[0031] a) Pretreatment: Take dried edible mushrooms, crush them by ultra-fine wall breaking to obtain ultra-fine mushroom powder, add distilled water in a mixing tank at a material-liquid ratio of 1:10-20, stir and dissolve to obtain mushroom solution;

[0032] b) High-voltage pulse electric field treatment: The mushroom solution in the preparation tank is sent to the pulse electric field treatment chamber through a peristaltic pump for high-voltage pulse electric field treatment, with the electric field intensity controlled at 10-20 kV / cm and the number of pulses at 6-14;

[0033] c) Protein extraction: add sodium hydroxide solution to the mushroom liquid treated with high-voltage pulse electric field to adjust the pH to 8.0, let it stand for 5-15 minutes, then add hydrochloric acid solution to adjust the pH to 4.5, centrifuge for 5-15 minutes and collect the precipitate;

[0034] d) compound enzyme hydrolysis: putting the precipitate into an enzymolysis tank, adding distilled water to prepare a 1%-5% solution, adjusting the pH to 8.0, heating to 40±0.5° C., and adding 3000-7000 U / g of compound enzyme for 1 to 3 hours of enzymolysis, wherein the compound enzyme comprises at least two proteases selected from the group consisting of papain, alkaline protease, neutral protease, pepsin, trypsin, cathepsin and subtilisin;

[0035] e) collecting the supernatant: heating the enzymatically hydrolyzed liquid to 85-95° C. and maintaining the temperature for 5-15 minutes, cooling the liquid and centrifuging at 4000-6000 r / min for 10-20 minutes to collect the supernatant; and

[0036] f) Filtration: Filter the supernatant through a filter membrane with a molecular weight ≥ 3KD to obtain a mushroom small molecule peptide concentrate.

[0037] In some embodiments, the edible mushroom used in the present invention is a genus of Pleurotus. Pleurotus is a genus in Fungi, Basidiomycetes, Hymeromycetes, Asterotomycetes, Agaricales, and Tricholomataceae, and is rich in polysaccharides, proteins, amino acids, fatty acids and other bioactive substances. More preferably, the edible mushroom is Pleurotus eryngii. Pleurotus eryngii is a typical edible and medicinal fungus with bright color, delicious taste, rich in nutrients such as protein, amino acids, trace elements, polysaccharides, and has multiple biological activities such as anti-aging and anti-oxidation. The preparation process of the present invention is particularly suitable for selecting Pleurotus eryngii as an edible mushroom substrate, so that small molecule peptides with higher nutritional content and better peptide molecule distribution can be obtained.

[0038] The aforementioned step a) is pretreatment, that is, pretreatment of the edible mushroom substrate to obtain a mushroom solution through the ultrafine mushroom powder. In this step, the ultrafine mushroom powder is added to distilled water in a mixing tank at a solid-liquid ratio of 1:10-20, preferably 1:15-20, and stirred to dissolve to obtain a mushroom solution.

[0039] The above step b) is a high-voltage pulse electric field treatment. The high-voltage pulse electric field sterilization technology is to generate an instantaneous high-voltage pulse electric field between two electrodes to act on food, and is widely used in food sterilization, wine aging, material extraction, etc. Under the action of the high-voltage pulse electric field, the charge distribution on the cell membrane changes, resulting in an increase in the potential difference between the two surfaces of the cell membrane. When this potential difference reaches or exceeds the critical potential of the cell membrane, the cell membrane will form a perforation, and the size and reversibility of the pore size depend on the thickness, shape and intensity of the cell membrane. Irreversible perforation will cause the cell contents (such as proteins, ions, etc.) to leak, resulting in loss of cell function. In the preparation process of the present invention, the mushroom solution obtained in the above step a) is sent to the pulse electric field treatment chamber by a peristaltic pump for high-voltage pulse electric field treatment, and the electric field strength is controlled to be 10-20kV / cm and the number of pulses is 6-14. The above-mentioned high-voltage pulse electric field parameters are specially optimized for edible mushroom substrates, and thus can promote the large-scale and rapid dissolution of edible mushroom cell contents.

[0040] In some preferred embodiments, the flow rate of the peristaltic pump controlled in step b) is 0.8-1.2 L / min, the electric field strength is 14-16 kV / cm, and the number of pulses is 8-12. More preferably, the flow rate of the peristaltic pump controlled is 1 L / min, the electric field strength is 15 kV / cm, and the number of pulses is 10. The applicant was surprised to find that for edible mushroom substrates, especially based on the speed at which its cell contents seep out during high-voltage pulse electric field treatment, the various parameters (electric field strength and number of pulses) of the high-voltage pulse electric field and the flow rate of the peristaltic pump all need to be specially set to avoid excessive electric field strength destroying the components in the cells or too small electric field strength being insufficient to destroy the cells. Under the above parameters preferred by the present invention, the high-voltage pulse electric field will achieve a better treatment effect for edible mushrooms.

[0041] The aforementioned step c) is protein extraction, and sodium hydroxide solution and hydrochloric acid solution are added to the mushroom liquid after high-voltage pulse electric field treatment, and the pH values ​​are adjusted to the corresponding values, so as to obtain mushroom protein by precipitating. Preferably, 0.1 mol / L sodium hydroxide solution is used to adjust the pH to 8.0, and the mixture is allowed to stand for 5-15 minutes, preferably 5-10 minutes, and then 0.1 mol / L hydrochloric acid solution is used to adjust the pH to 4.5, and the precipitate is obtained after centrifugation for 5-15 minutes, preferably 10-15 minutes. The aforementioned centrifugal speed can be any suitable speed, preferably 4000-6000 r / min, and more preferably 5000 r / min.

[0042] The aforementioned step d) is a composite enzyme hydrolysis, wherein the aforementioned centrifugal precipitate is placed in an enzymolysis tank, distilled water is added to prepare a 1%-5% solution, the pH is adjusted to 8.0, heated to 40±0.5°C, and 3000-7000U / g of composite enzyme is added, and the enzymolysis time is 1 to 3 hours. In the present invention, the composite enzyme comprises at least two proteases, preferably at least three proteases. The protease is selected from the group consisting of papain, alkaline protease, neutral protease, pepsin, trypsin, cathepsin and subtilisin. More preferably, the protease is selected from the group consisting of papain, alkaline protease, neutral protease, which are specially selected based on the protein type of the edible mushroom substrate, thereby improving the productivity of the peptide and optimizing the distribution of peptide molecules.

[0043] In a highly preferred embodiment, the composite enzyme comprises at least two of papain, alkaline protease, and neutral protease, and more preferably a combination of the three proteases, i.e., papain, alkaline protease, and neutral protease. When two of the aforementioned proteases are included, the two proteases preferably reach a specific ratio in the composite enzyme, such as the ratio of papain to alkaline protease is 1:(1.5-2), the ratio of papain to neutral protease is 1:(1-1.5), and the ratio of alkaline protease to neutral protease is (1.5-2):(1-1.5). When three of the aforementioned proteases are included, preferably the three proteases reach a specific ratio in the composite enzyme, more preferably 1:(1.5-2):(1-1.5).

[0044] In other embodiments, the amount of the compound enzyme added is 4000-6000 U / g, more preferably 5000 U / g, and the enzymolysis time is 1.5-2.5 hours. As mentioned above, the traditional multi-step enzymatic method is usually time-consuming, at least 5-10 hours, so the compound enzyme method of the present invention is more suitable for high-efficiency, large-scale industrial production.

[0045] The aforementioned step e) is to collect the supernatant, and the above enzymatically hydrolyzed liquid is heated, then cooled and centrifuged, and the supernatant is collected. In some embodiments, the enzymatically hydrolyzed liquid is heated to 85-95°C, and maintained for 5-10 minutes, preferably heated to 90°C, and maintained for 10 minutes. After the liquid is cooled, it is centrifuged at 4000-6000r / min for 10-20 minutes, preferably at 5000r / min for 15 minutes, and the supernatant is collected.

[0046] The aforementioned step f) is a filtration step, wherein the supernatant obtained in the aforementioned step e) is filtered to obtain a concentrated solution of mushroom small molecule peptides. The molecular weight of the filter membrane used for filtration is ≥3KD, preferably ≥2KD. In some embodiments, multiple filtrations may be used, such as sequentially passing through a microfiltration membrane, an ultrafiltration membrane, and a nanofiltration membrane, and finally cutting off a concentrated solution with a molecular weight of 2KD.

[0047] In some preferred embodiments, the edible mushroom small molecule peptide preparation process further comprises step g) freeze drying: freeze drying the mushroom small molecule peptide concentrate obtained in the above step f) to form a dry powder to obtain edible mushroom small molecule peptide dry powder. The freeze drying step can be carried out by any conventional freeze drying method.

[0048] Preparation device

[0049] In conjunction with the preparation process of the present invention, the present invention provides a device for preparing edible mushroom small molecule peptides, comprising:

[0050] A mixing tank is used to contain mushroom powder and distilled water to dissolve and obtain a mushroom solution;

[0051] a peristaltic pump, the inlet of which is connected to the mixing tank, and is used to deliver the mushroom solution from the mixing tank to the electric pulse generator;

[0052] The electric pulse generator is connected to the outlet of the peristaltic pump and is used to perform high-voltage pulse electric field treatment on the mushroom solution;

[0053] A centrifugal device, used for centrifuging the solution after protein extraction;

[0054] Enzyme hydrolysis tank, used for compound enzyme hydrolysis reaction.

[0055] It will be appreciated by those skilled in the art that the various devices used in the preparation device of the present invention may be conventional devices in the art, as long as the relevant parameters thereof meet the specific requirements of the present invention and are suitable for the preparation process of the present invention. For example, a peristaltic pump is a liquid delivery device capable of controlling the flow rate, and is widely used in the fields of scientific research, pharmaceuticals, chemicals, and food. In the present invention, based on the edible mushroom substrate, in particular, based on the rate at which its cell contents seep out during high-voltage pulsed electric field treatment, specific requirements are placed on the flow rate of the peristaltic pump, so as to achieve optimal production efficiency. Therefore, any peristaltic pump may be used in the preparation device of the present invention, as long as its controllable flow rate is within the range of 0.8-1.2 L / min required by the preparation process of the present invention.

[0056] Mushroom small molecule peptides and their applications

[0057] The mushroom small molecule peptide prepared by the present invention is a concentrated liquid or a dry powder. Specifically, according to the process for preparing edible mushroom small molecule peptides of the present invention, the obtained in step f) is a concentrated liquid of mushroom small molecule peptides; preferably, the obtained in step g) is a dry powder of edible mushroom small molecule peptides. Depending on the final application field, a suitable form of mushroom small molecule peptides can be selected.

[0058] In some embodiments, the mushroom small molecule peptide prepared by the present invention is a dry powder, wherein the content of peptides with a molecular weight of <1kD is >80%, and preferably the content of peptides with a molecular weight of <1kD in the mushroom small molecule peptide dry powder is >85%.

[0059] The mushroom small molecule peptide of the present invention can be applied to any suitable field, such as cosmetics, health products, medicines, etc. In one embodiment, the present invention provides cosmetics comprising the mushroom small molecule peptide, preferably the mushroom is a Pleurotus genus fungus, more preferably Pleurotus aurantii. It is worth noting that since Pleurotus aurantii is rich in various nutrients, its small molecule peptide can achieve better anti-aging, anti-oxidation and whitening effects when applied to cosmetics.

[0060] Peptide content detection method

[0061] The edible mushroom small molecule peptide prepared by the present invention can be detected by any conventional peptide content detection method, preferably by the method in Appendix A of GB / T 22729-2008, to detect the content of peptides with a molecular weight of less than 1 kD. This standard is included in the national standard full text disclosure system, see https: / / openstd.samr.gov.cn / bzgk / gb / newGblnfo?hcno=0010CAD4D17092E1B64A1742DBA101B4. DETAILED DESCRIPTION

[0062] The examples herein are intended to illustrate the invention but are not intended to limit or otherwise define the scope of the invention.

[0063] Examples 1-3 are examples of the preparation process of the edible mushroom small molecule peptide of the present invention, and comparative examples 4-5 are comparative preparation methods. Example 6 is the peptide content detection results of each example and comparative example, and Example 7 is the application effect experimental data of each example, comparative example and control group.

[0064] Example 1: Preparation process of Pleurotus eryngii small molecule peptide

[0065] (1) Pretreatment: Take dried Pleurotus eryngii mushrooms, crush them by ultrafine wall breaking and set aside, put 15 times of pure water in a mixing tank, then add ultrafine mushroom powder, stir and dissolve;

[0066] (2) High-voltage pulse electric field treatment: The peristaltic pump flow rate was adjusted to 1 L / min to pump the solution in the preparation tank to the pulse electric field treatment chamber for high-voltage pulse electric field treatment. The electric field intensity was controlled to be 15 kV / cm and the number of pulses was 10.

[0067] (3) Protein extraction: The treated mushroom liquid was adjusted to pH 8.0 with 0.1 mol / L sodium hydroxide solution, allowed to stand for 5 min, then adjusted to pH 4.5 with 0.1 mol / L hydrochloric acid solution, centrifuged at 5000 r / min for 15 min, and the precipitate was placed in an enzymatic hydrolysis tank and distilled water was added to prepare a 2% solution.

[0068] (4) Compound enzyme hydrolysis: adjust the pH of the solution obtained in (3) to 8.0, heat to 40°C, add compound enzyme, which is papain and alkaline protease in a ratio of 1:2, add 5000U / g enzyme, and perform enzymatic hydrolysis for 1.5h.

[0069] (5) Heat the enzymatically hydrolyzed liquid to 90°C and maintain for 10 min to inactivate the enzyme.

[0070] (6) After the liquid is cooled, centrifuge at 5000 r / min for 15 min and collect the supernatant.

[0071] (7) The supernatant is passed through a microfiltration membrane, an ultrafiltration membrane, and a nanofiltration membrane in sequence, and finally a concentrate with a molecular weight cutoff of 2KD is obtained.

[0072] (8) freeze-drying the concentrated solution to form a dry powder, thereby obtaining Pleurotus eryngii peptide powder.

[0073] Example 2: Preparation process of Pleurotus eryngii small molecule peptide

[0074] The preparation process of Example 2 is the same as that of Example 1, the only difference being that in step (4) the composite enzyme hydrolysis, the composite enzyme used is papain, alkaline protease and neutral protease, and the ratio of the three proteases in the composite enzyme is 1:1.5:1.

[0075] Example 3: Preparation process of Pleurotus eryngii small molecule peptide

[0076] The preparation process of Example 3 is the same as that of Example 1, the only difference being that in step (4) the composite enzyme hydrolysis, the composite enzyme used is papain, alkaline protease and neutral protease, and the ratio of the three proteases in the composite enzyme is 1:2:1.

[0077] Comparative Example 4:

[0078] The preparation process of Comparative Example 4 is the same as that of Example 3, with the only difference being that step (2) high-voltage pulse electric field treatment is missing, and the remaining steps are the same.

[0079] Comparative Example 5:

[0080] The preparation process of Comparative Example 5 is the same as that of Example 3, with the only difference being that the enzyme added in step (4) is only papain, the enzyme amount added is 5000 U / g, the enzymatic hydrolysis time is 1.5 h, and the remaining steps are the same.

[0081] Example 6: Peptide content detection data

[0082] According to the method in Appendix A of GB / T 22729-2008, the content of peptides with a molecular weight of less than 1000 Da in each example and comparative example was detected, and the obtained peptide contents are shown in Table 1:

[0083] Table 1

[0084] serial number Example 1 Example 2 Example 3 Comparative Example 4 Comparative Example 5 Peptide content% 84.57 87.72 89.35 71.21 77.28

[0085] Example 7: Application effect test data

[0086] The peptide powders obtained in the above Examples 1-3 and Comparative Examples 4-5 were made into solutions with a concentration of 2%, and poured into a spray bottle for later use.

[0087] Subject selection: 60 female volunteers aged 35 to 50 were randomly selected and divided into 6 groups, with 10 subjects in each group. The sprays of Examples 1-3 and Comparative Examples 4-5, as well as pure water spray were used as the control group.

[0088] Test method: Spray on face and arms once in the morning and evening. Clean the skin before use. Record the results of continuous use for 15 days and 30 days. No other skin care products should be used on the test area during use.

[0089] Judgment criteria: The skin color is divided into 10 levels, with the lightest color being level 1 and the darkest being level 10. The evaluation is mainly based on the subjective evaluation of the volunteers, and the corresponding evaluation results (no obvious effect, slight effect or significant effect) are classified according to the level of improvement of the skin on the hands and face. See Table 2 for specific judgment criteria.

[0090] Test results: According to the evaluation results of each group of subjects, the number of people who achieved each evaluation result was counted. The results are shown in Table 3.

[0091] Table 2

[0092]

[0093] Table 3

[0094]

[0095]

[0096] From the above results, it can be seen that the spray made with the small molecule peptides of Pleurotus eryngii prepared in Examples 1-3 has obvious skin care effects and can achieve whitening and anti-aging effects. In contrast, the small molecule peptides prepared in Comparative Example 4 have not been treated with a high-voltage electric field and have no obvious whitening and anti-aging effects; Comparative Example 5 only uses one enzyme, and the whitening and skin care effects are poor.

[0097] Although specific embodiments of the present invention have been described and illustrated, it will be appreciated by those skilled in the art that various other changes and modifications may be made without departing from the spirit and scope of the present invention. Therefore, all such changes and modifications are intended to fall within the scope of the claims of this application.

[0098] All percentages, ratios and proportions are calculated by weight of the total composition unless otherwise indicated. All temperatures are in degrees Celsius (° C.) unless otherwise indicated. All component or composition levels refer to the active level of that component or composition, and are exclusive of impurities, such as residual solvents or by-products, which may be present in commercially available sources.

[0099] It should be understood that each numerical upper limit given in this specification includes values ​​lower than it, just as if these lower values ​​are explicitly written in the application. Each numerical lower limit given in this specification includes values ​​higher than it, just as if these higher values ​​are explicitly written in the application. Each numerical range given in this specification includes narrower numerical ranges falling within the numerical range, just as if these narrower numerical ranges are explicitly written in the application.

[0100] The values ​​and dimensions disclosed herein should not be understood as being strictly limited to the exact values ​​recited. Instead, unless otherwise specified, each such value and dimension is intended to represent the recited value and a functionally equivalent range around that value. For example, a dimension disclosed as "40 millimeters" is intended to mean "about 40 millimeters."

[0101] As used herein, the term "mixing" refers to adding ingredients together and achieving homogeneity, and the term "mixture" refers to a homogeneous mixture of ingredients.

[0102] In the present application, when a composition is “substantially free” of a specific ingredient, it means that the composition contains less than a trace amount, or less than 0.1%, or less than 0.01%, or less than 0.001%, based on the mass of the composition, of the specific ingredient.

[0103] In this application, unless the order is particularly clear, the terms "a)", "b)" and "c)" used to refer to the steps in the preparation method do not indicate the order. For example, step a) may occur before step b) or after step b), or steps b) and c) may occur simultaneously, or step b) may occur after step c).

[0104] The terms "comprising", "including", "comprising", "including", "comprising", "including", "containing", "including" and "containing" used in this application are intended to be non-limiting, that is, other ingredients and other steps that do not affect the end result may be added.

Claims

1. A process for preparing a small molecule peptide of edible mushrooms, comprising: a) Pretreatment: Take dried edible mushrooms, crush them by ultra-fine wall breaking to obtain ultra-fine mushroom powder, add distilled water in a mixing tank at a material-liquid ratio of 1:10-20, stir and dissolve to obtain mushroom solution; b) High-voltage pulse electric field treatment: The mushroom solution in the preparation tank is sent to the pulse electric field treatment chamber through a peristaltic pump for high-voltage pulse electric field treatment, with the electric field intensity controlled at 10-20 kV / cm and the number of pulses at 6-14; c) Protein extraction: add sodium hydroxide solution to the mushroom liquid treated with high-voltage pulse electric field to adjust the pH to 8.0, let it stand for 5-15 minutes, then add hydrochloric acid solution to adjust the pH to 4.5, centrifuge for 5-15 minutes and collect the precipitate; d) compound enzyme hydrolysis: placing the precipitate in an enzymolysis tank, adding distilled water to prepare a 1%-5% solution, adjusting the pH to 7-8, heating to 40±0.5° C., and adding 3000-7000 U / g of compound enzyme for 1 to 3 hours of enzymolysis, wherein the compound enzyme comprises at least two proteases selected from the group consisting of papain, alkaline protease, neutral protease, pepsin, trypsin, cathepsin and subtilisin; e) collecting the supernatant: heating the enzymatically hydrolyzed liquid to 85-95° C. and maintaining the temperature for 5-15 minutes, cooling the liquid and centrifuging at 4000-6000 r / min for 10-20 minutes to collect the supernatant; and f) Filtration: Filter the supernatant through a filter membrane with a molecular weight ≥ 3KD to obtain a mushroom small molecule peptide concentrate.

2. The process for preparing the edible mushroom small molecule peptide according to claim 1, further comprising: g) Freeze drying: freeze-drying the mushroom small molecule peptide concentrate obtained in step f) to form a dry powder, thereby obtaining edible mushroom small molecule peptide dry powder.

3. The process for preparing small molecule peptides from edible mushrooms according to claim 1, wherein in step b), the flow rate of the peristaltic pump is controlled to be 0.8-1.2 L / min, the electric field strength is controlled to be 14-16 kV / cm, and the number of pulses is controlled to be 8-12.

4. The process for preparing small molecule peptides of edible mushrooms according to claim 1, wherein in step d), the composite enzyme comprises papain, alkaline protease and neutral protease, and the ratio of the three proteases in the composite enzyme is 1:(1.5-2):(1-1.5).

5. The process for preparing small molecule peptides from edible mushrooms according to claim 4, wherein in the step d), the amount of the complex enzyme added is 4000-6000 U / g, and the enzymolysis time is 1.5-2.5 hours.

6. The process for preparing small molecule peptides from edible mushrooms according to claim 1, wherein the edible mushrooms are Pleurotus eryngii.

7. A device for preparing small molecule peptides of edible mushrooms, comprising: A mixing tank is used to contain mushroom powder and distilled water to dissolve and obtain a mushroom solution; a peristaltic pump, the inlet of which is connected to the mixing tank, and is used to deliver the mushroom solution from the mixing tank to the electric pulse generator; The electric pulse generator is connected to the outlet of the peristaltic pump and is used to perform high-voltage pulse electric field treatment on the mushroom solution; A centrifugal device, used for centrifuging the solution after protein extraction; and Enzyme hydrolysis tank, used for compound enzyme hydrolysis reaction.

8. The mushroom small molecule peptide prepared by the process for preparing edible mushroom small molecule peptide according to any one of claims 1 to 6, wherein the mushroom small molecule peptide is a concentrated solution or a dry powder.

9. The mushroom small molecule peptide according to claim 8, wherein the mushroom small molecule peptide is in the form of dry powder, wherein the content of peptides with a molecular weight of less than 1 kD is greater than 80%.

10. A cosmetic comprising the mushroom small molecule peptide according to claim 9.

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