A small molecule peptide composition prepared by fermenting Cordyceps sinensis, its application, and cosmetics

Abstract

This application belongs to the technical field of cosmetic production, and discloses a small molecule peptide composition prepared by fermenting Cordyceps sinensis. The small molecule peptide composition prepared by fermenting Cordyceps sinensis is a small molecule peptide composition prepared by fermenting Cordyceps sinensis containing polypeptides. The small molecule peptide composition prepared by fermenting Cordyceps sinensis containing polypeptides is prepared by mixing the fermentation product and the lysate obtained by fermenting Cordyceps sinensis with a galactose yeast fermentation broth. The mass ratio of the galactose yeast fermentation broth to Cordyceps sinensis is 75-95:2-10. Through the above design, this application obtains a small molecule peptide composition prepared by fermenting Cordyceps sinensis with good free radical scavenging ability, antioxidant enzyme activation ability, and cell repair promotion ability. In addition, the antioxidant ability of the composition is further enhanced by fermenting the raw materials with galactose yeast. Furthermore, this application also discloses the use of the above-mentioned small molecule peptide composition prepared by fermenting Cordyceps sinensis and a cosmetic.

Description

Technical Field

[0001] This application relates to the technical field of cosmetics production, and particularly to a small molecule peptide composition prepared by fermenting Cordyceps sinensis, its application, and cosmetics. Background Art

[0002] Antioxidation and anti-aging are of great significance to the skin because the skin is the largest organ of the human body and the part most directly exposed to the external environment. This makes the skin vulnerable to external attacks, such as ultraviolet rays, pollutants, free radicals in the air, etc. These external attacks can cause oxidative damage to skin cells and accelerate the process of skin aging.

[0003] Oxidation refers to a chemical reaction in which molecules lose electrons, creating highly reactive molecules called free radicals. Free radicals can damage proteins, lipids, and DNA within cells through oxidation, resulting in impaired cell function. The accumulation of such oxidative damage accelerates the aging process of the skin, causing aging manifestations such as wrinkles, sagging, and age spots on the skin.

[0004] The role of antioxidants is to neutralize and scavenge free radicals in the body, prevent the occurrence of oxidation reactions, and thus slow down the aging problems caused by oxidative damage. In addition, the significance of anti-aging lies in delaying the process of skin aging and maintaining the youthful state of the skin.

[0005] Prior Art 1: Chinese Patent Application No. 202010764419.X discloses a filtrate of bifidobacterium fermentation product and its application in skin care products. The filtrate of bifidobacterium fermentation product is prepared by the following method:

[0006] (1) Take Cordyceps militaris and Cyanotis arachnoidea, mix and crush them to obtain a mixed raw material; add water to the mixed raw material, crush, juice, centrifuge, and take the supernatant;

[0007] (2) Add glucose, soluble starch, agar, L-cysteine, yeast extract powder, potassium dihydrogen phosphate, peptone, beef extract powder, anhydrous sodium acetate, and tomato extract powder to the supernatant, stir evenly and sterilize to obtain a culture medium;

[0008] (3) Inoculate the culture medium with bifidobacterium strains, ferment to obtain a fermentation broth; adjust the pH value of the fermentation broth to 5.5 - 6.5, and perform high-pressure homogenization 3 times to fully break the yeast cells; filter to obtain a filtrate, that is, the filtrate of bifidobacterium fermentation product;

[0009] The filtrate of the bifida ferment lysate disclosed by this solution contains rich active small molecules such as vitamins, minerals, amino acids, polysaccharides, and polypeptides, and is a high-quality yeast essence only used for skin care. The nutrients it is rich in have the function of nourishing the skin. Applying it in skin care products can strengthen skin metabolism, promote cell proliferation, tighten the skin, reduce fine lines, and has a good anti-wrinkle function.

[0010] Prior Art 2: Chinese Patent Application No. 200510016899.7 discloses a cordyceps militaris polypeptide nano series skin care cosmetics and its preparation method. The skin care cosmetics are composed of a main agent, a matrix, and water. The main agent is a cordyceps militaris milk concentrate, and its dosage accounts for 2-10% by mass percentage. The main agent contains bioactive substances such as polypeptides, amino acids, and D-mannitol. The preparation method is to inoculate cordyceps militaris strains into a sterile skim milk medium, prepare cordyceps militaris milk concentrate through protease hydrolysis and ultramicroscopic treatment, and finally mix, emulsify, and sterilize to obtain the skin care cosmetics. The protein in the cordyceps militaris milk fermentation broth is hydrolyzed into small molecule bio-peptides, which can increase the skin moisturizing and skin care and beauty effects of cordyceps militaris milk. The treatment by the concentration and ultramicroscopic process is beneficial to the rapid penetration of bioactive factors into the skin and the exertion of their effects. According to different needs, adding the cordyceps militaris milk concentrate to the matrix can conveniently prepare skin care cosmetics of different dosage forms and different effects.

[0011] The problem to be solved by this solution: How to develop a small molecule peptide composition prepared by fermenting cordyceps sinensis that is different from the prior art and has good antioxidant capacity. Summary of the Invention

[0012] The purpose of this application is to provide a small molecule peptide composition prepared by fermenting cordyceps sinensis. The cordyceps sinensis extract used as the fermentation raw material has rich active ingredients such as proteins, polysaccharides, amino acids, and trace elements, and has strong antioxidant effects. It can scavenge free radicals in the body, reduce cell damage caused by oxidative stress, thereby delaying cell aging, improving the antioxidant capacity of the body, and further enhancing the antioxidant capacity of the composition through the fermentation of galactose yeast.

[0013] To achieve the above purpose, this application discloses a small molecule peptide composition prepared by fermenting cordyceps sinensis. The small molecule peptide composition prepared by fermenting cordyceps sinensis is a small molecule peptide composition prepared by fermenting cordyceps sinensis containing polypeptides;

[0014] The small molecule peptide composition prepared by fermenting cordyceps sinensis containing polypeptides is prepared by mixing the fermentation products and cell lysates obtained by fermenting cordyceps sinensis with a galactose yeast fermentation broth. The mass ratio of the galactose yeast fermentation broth to cordyceps sinensis is 75-95:2-10.

[0015] Cordyceps sinensis contains rich active ingredients such as proteins, polysaccharides, amino acids and trace elements. These ingredients have strong antioxidant effects, can scavenge free radicals in the body, reduce cell damage caused by oxidative stress, thus delaying cell aging and improving the body's antioxidant capacity. At the same time, active ingredients such as polysaccharides and amino acids in Cordyceps sinensis can regulate the metabolic process in the body, promote metabolism, reduce the accumulation of waste in the body, and slow down the rate of cell aging.

[0016] The filtrate obtained after fermentation with Saccharomyces cerevisiae var. galactosidase contains rich active substances such as polypeptides, enzymes, amino acids and antioxidant substances. These ingredients can effectively scavenge free radicals in the body, reduce cell damage caused by oxidative stress, thus achieving an antioxidant effect. The filtrate of Saccharomyces cerevisiae var. galactosidase fermentation products can also activate the antioxidant enzyme system in the body, such as superoxide dismutase, catalase, etc., which can enhance the antioxidant capacity of cells, reduce oxidative damage, and delay the process of cell aging. And the active ingredients of the filtrate of Saccharomyces cerevisiae var. galactosidase fermentation products can regulate the expression of a series of genes, affect the intracellular signal transduction pathway, promote cell growth, regeneration and repair functions, enhance the metabolic vitality of cells, and the synergistic effect of the two can further scavenge free radicals, activate antioxidant enzymes, and promote cell repair, keeping cells in a healthy state.

[0017] Preferably, the small molecule peptide composition prepared by fermenting Cordyceps sinensis is prepared by mixing the fermentation products and lysed products obtained by fermenting Cordyceps sinensis and Tremella fuciformis with Saccharomyces cerevisiae var. galactosidase fermentation broth, and the mass ratio of the Saccharomyces cerevisiae var. galactosidase fermentation broth to Cordyceps sinensis and Tremella fuciformis is 75-95:2-10:1-3.

[0018] Preferably, the preparation method of the small molecule peptide composition prepared by fermenting Cordyceps sinensis is specifically as follows:

[0019] Step 1: Dry and crush Cordyceps sinensis to obtain powder.

[0020] Step 2: Mix the powder obtained in Step 1 with pure water, yeast powder and disodium hydrogen phosphate to obtain a culture medium.

[0021] Step 3: Inoculate the Saccharomyces cerevisiae var. galactosidase fermentation broth into the culture medium and ferment for 4-8 h, then sterilize at high temperature, crush and centrifuge to obtain the small molecule peptide composition prepared by fermenting Cordyceps sinensis containing fermentation products and lysed products.

[0022] Preferably, Step 1 is specifically as follows:

[0023] Dry and crush Cordyceps sinensis and Tremella fuciformis to obtain powder, and the mass ratio of Cordyceps sinensis to Tremella fuciformis is 2-10:1-3.

[0024] Preferably, the powder also includes grains, and Step 1 is specifically as follows:

[0025] Cordyceps sinensis, Tremella fuciformis and grains are dried and crushed to obtain powders, wherein the mass ratio of galactomyces to Cordyceps sinensis, Tremella fuciformis and grains is 2-10:1-3:1-3:

[0026] The grains are selected from at least one of wheat, barley, rye, oats and buckwheat.

[0027] Preferably, the grains are wheat, rye and buckwheat, and the mass ratio of wheat, rye and buckwheat is 2:1-2:3.

[0028] In addition, the present application also discloses the use of the small molecule peptide composition prepared by fermenting Cordyceps sinensis as described above for preparing cosmetics.

[0029] In addition, the present application also discloses a cosmetic containing 0.1-100 wt% of the small molecule peptide composition prepared by fermenting Cordyceps sinensis as described above.

[0030] Preferably, it further includes a solvent, and the solvent is selected from at least one of glycerol and butanediol.

[0031] Preferably, the dosage form of the cosmetic is aqueous solution, essence, mask liquid, emulsion or cream.

[0032] The beneficial effects of the present application are:

[0033] In the small molecule peptide composition prepared by fermenting Cordyceps sinensis disclosed in the present application, Cordyceps sinensis contains rich active ingredients, such as proteins, polysaccharides, amino acids and trace elements, etc. These ingredients have strong antioxidant effects, can scavenge free radicals in the body, reduce cell damage caused by oxidative stress, so as to delay cell aging and improve the antioxidant capacity of the body. At the same time, active ingredients such as polysaccharides and amino acids in Cordyceps sinensis can regulate the metabolic process in the body, promote metabolism, reduce the accumulation of waste in the body, and slow down the speed of cell aging;

[0034] The filtrate obtained after fermentation with galactomyces contains rich active substances, such as polypeptides, enzymes, amino acids and antioxidant substances, etc. These ingredients can effectively scavenge free radicals in the body, reduce cell damage caused by oxidative stress, so as to achieve the antioxidant effect. The filtrate of galactomyces fermentation products can also activate the antioxidant enzyme system in the body, such as superoxide dismutase, catalase, etc., which can enhance the antioxidant capacity of cells, reduce oxidative damage, and delay the process of cell aging. And the active ingredients of the filtrate of galactomyces fermentation products can regulate the expression of a series of genes, affect the intracellular signal transduction pathway, promote the growth, regeneration and repair functions of cells, enhance the metabolic vitality of cells, and the synergistic effect of the two can further scavenge free radicals, activate antioxidant enzymes, and promote cell repair, so that the cells remain in a healthy state. Description of the Drawings

[0035] Figure 1 Schematic diagram of a chicken embryo before adding the solution prepared from the small molecule peptide composition prepared by fermenting Cordyceps sinensis in Example 1;

[0036] Figure 2 Schematic diagram of a chicken embryo after adding the solution prepared from the small molecule peptide composition prepared by fermenting Cordyceps sinensis in Example 1;

[0037] Figure 3 Chromatogram of the small molecule peptide composition prepared by fermenting Cordyceps sinensis prepared in Example 1;

[0038] Figure 4 Mass spectrum of the small molecule peptide composition prepared by fermenting Cordyceps sinensis prepared in Example 1;

[0039] Figure 5 Structural diagram of the bioactive polypeptide of the small molecule peptide composition prepared by fermenting Cordyceps sinensis prepared in Example 1. Detailed implementation manners

[0040] The present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. In the description of the present invention, it should be noted that for those embodiments not specified with specific conditions, they are carried out according to conventional conditions or conditions recommended by the manufacturer. For reagents or instruments not specified with the manufacturer, they are all conventional products that can be obtained through commercial purchase.

[0041] Before presenting the embodiments, the following necessary explanations are made on the preparation and acquisition routes of the raw materials involved in the embodiments:

[0042] The preservation number of Saccharomyces galactomyces is GDMCC No: 64235, the preservation date is January 2, 2024; the preservation unit is Guangdong Microbial Culture Collection Center; the preservation address is the 5th floor of Building 59, No. 100, Xianlie Middle Road, Guangzhou, and the Latin name is Geotrichum candidum .

[0043] Cordyceps sinensis was purchased from Ningbo Solangdun Co., Ltd.;

[0044] Tremella fuciformis was purchased from Honghe Agricultural Development Co., Ltd. in Gutian County;

[0045] Wheat was purchased from Pingshuo Trading;

[0046] Barley was purchased from Pingshuo Trading;

[0047] Rye was purchased from the Vegetarian Fun Flagship Store;

[0048] Buckwheat was purchased from the Huantai Kangjian Food Franchise Store;

[0049] The yeast powder was purchased from Guangzhou Dingguo Biotechnology Co., Ltd., 500 g / bottle, brand OXOID, model LP0021B.

[0050] It should be noted that, unless otherwise specified, the galactose yeast fermentation broth used in the examples and comparative examples was prepared by fermenting the culture medium with the galactose yeast with the preservation number of GDMCC No: 64235, the preservation date of January 2, 2024; the preservation unit was Guangdong Microbial Culture Collection Center; the preservation address was on the 5th floor of Building 59, No. 100 compound, Xianlie Middle Road, Guangzhou, and the Latin name was Geotrichum candidum fermented at 28 °C for 24 h; and the mass ratio of galactose yeast to the culture medium was 3:50;

[0051] Meanwhile, the culture medium was prepared by mixing wheat flour obtained by drying and crushing wheat with yeast powder, sodium dihydrogen phosphate, and pure water. More specifically, the mass fraction of wheat flour in the culture medium was 5%, the mass fraction of yeast powder was 2%, the mass fraction of sodium dihydrogen phosphate was 1%, and the mass fraction of pure water was 92%.

[0052] Example 1

[0053] Step 1: Dry the Cordyceps sinensis at 60 °C for 6 h and crush it to obtain a powder;

[0054] Step 2: Mix the powder obtained in Step 1 with pure water, yeast powder, and sodium dihydrogen phosphate to obtain a culture medium, and in the culture medium, the mass fraction of yeast powder was 2%, the mass fraction of sodium dihydrogen phosphate was 1%, the mass fraction of pure water was 92%, and the mass fraction of the powder was 5%;

[0055] Step 3: Inoculate the galactose yeast fermentation broth into the culture medium and ferment for 6 h, then sterilize at 121 °C, crush, centrifuge, and take the supernatant to obtain a small molecule peptide composition prepared by fermenting Cordyceps sinensis containing fermentation products and cell lysate products;

[0056] And the mass ratio of the galactose yeast fermentation broth in Step 3 to the powder in Step 1 was 87.5:5.

[0057] Example 2

[0058] Step 1: Dry the Cordyceps sinensis at 60 °C for 6 h and crush it to obtain a powder;

[0059] Step 2: Mix the powder obtained in Step 1 with pure water, yeast powder, and sodium dihydrogen phosphate to obtain a culture medium, and in the culture medium, the mass fraction of yeast powder was 2%, the mass fraction of sodium dihydrogen phosphate was 1%, the mass fraction of pure water was 92%, and the mass fraction of the powder was 5%;

[0060] Step 3: Inoculate the galactose yeast fermentation broth into the culture medium and ferment for 4 h. Then, sterilize at 121 °C, break, centrifuge, and take the supernatant to obtain a small molecule peptide composition prepared by fermenting Cordyceps sinensis containing fermentation products and cell lysate products;

[0061] Moreover, the mass ratio of the galactose yeast fermentation broth in Step 3 to the powder in Step 1 is 95:2.

[0062] Example 3

[0063] Step 1: Dry Cordyceps sinensis at 60 °C for 6 h and break it to obtain a powder;

[0064] Step 2: Mix the powder obtained in Step 1 with pure water, yeast powder, and sodium dihydrogen phosphate to obtain a culture medium. The mass fraction of yeast powder in the culture medium is 2%, the mass fraction of sodium dihydrogen phosphate is 1%, the mass fraction of pure water is 92%, and the mass fraction of the powder is 5%;

[0065] Step 3: Inoculate the galactose yeast fermentation broth into the culture medium and ferment for 8 h. Then, sterilize at 121 °C, break, centrifuge, and take the supernatant to obtain a small molecule peptide composition prepared by fermenting Cordyceps sinensis containing fermentation products and cell lysate products;

[0066] Moreover, the mass ratio of the galactose yeast fermentation broth in Step 3 to the powder in Step 1 is 75:10.

[0067] Example 4

[0068] It is basically the same as Example 1, except that in Step 3, the mass ratio of the galactose yeast fermentation broth to the powder in Step 1 is 90.5:2.

[0069] Example 5

[0070] It is basically the same as Example 1, except that Step 1 is specifically: dry and break Cordyceps sinensis and Tremella fuciformis to obtain a powder, and the mass ratio of Cordyceps sinensis to Tremella fuciformis in the powder is 5:1;

[0071] Moreover, the mass ratio of the galactose yeast fermentation broth in Step 3 to the powder in Step 1 is 87.5:5.

[0072] Example 6

[0073] It is basically the same as Example 1, except that Step 1 is specifically: dry and break Cordyceps sinensis, Tremella fuciformis, and grains to obtain a powder, and the mass ratio of Cordyceps sinensis, Tremella fuciformis, and grains in the powder is 5:1:1;

[0074] The grain is wheat, and the mass ratio of the galactose yeast fermentation broth in Step 3 to the powder is 87.5:5.

[0075] Example 7

[0076] It is basically the same as Example 1, except that in Step 1, specifically: Cordyceps sinensis, Tremella fuciformis, and grains are dried and crushed to obtain a powder, and the mass ratio of Cordyceps sinensis, Tremella fuciformis, and grains in the powder is 5:1:1;

[0077] The grains are a mixture of barley and buckwheat, and the mass ratio of barley to buckwheat is 3.5:3.

[0078] And the mass ratio of the galactose yeast fermentation broth in Step 3 to the powder in Step 1 is 87.5:5.

[0079] Example 8

[0080] It is basically the same as Example 1, except that in Step 1, specifically: Cordyceps sinensis, Tremella fuciformis, and grains are dried and crushed to obtain a powder, and the mass ratio of Cordyceps sinensis, Tremella fuciformis, and grains in the powder is 5:1:1;

[0081] The grains are a mixture of rye and buckwheat, and the mass ratio of rye to buckwheat is 3.5:3;

[0082] And the mass ratio of the galactose yeast fermentation broth in Step 3 to the powder in Step 1 is 87.5:5.

[0083] Example 9

[0084] It is basically the same as Example 1, except that in Step 1, specifically: Cordyceps sinensis, Tremella fuciformis, and grains are dried and crushed to obtain a powder, and the mass ratio of Cordyceps sinensis, Tremella fuciformis, and grains in the powder is 5:1:1;

[0085] The grains are a mixture of wheat, rye, and buckwheat, and the mass ratio of wheat, rye, and buckwheat is 2:1.5:3.

[0086] And the mass ratio of the galactose yeast fermentation broth in Step 3 to the powder in Step 1 is 87.5:5.

[0087] Example 10

[0088] It is basically the same as Example 1, except that in Step 3, a commercially available galactose yeast is used to replace galactose yeast ( Geotrichum candidum ) to prepare the galactose yeast fermentation broth 。

[0089] Example 11

[0090] It is basically the same as Example 5, except that the same mass of Portulaca oleracea is used to replace Tremella fuciformis to obtain a powder, and the mass ratio of the galactose yeast fermentation broth in Step 3 to the powder in Step 1 is 87.5:5.

[0091] Example 12

[0092] Basically the same as Example 5, except that wolfberry of the same quality is used to replace tremella, obtaining a powder, and the mass ratio of the galactose yeast fermentation broth in Step 3 to the powder in Step 1 is 87.5:5.

[0093] Example 13

[0094] Basically the same as Example 5, except that sargassum thunbergii of the same quality is used to replace tremella, obtaining a powder, and the mass ratio of the galactose yeast fermentation broth in Step 3 to the powder in Step 1 is 87.5:5.

[0095] Comparative Example 1

[0096] Step 1: Dry and crush cordyceps sinensis at 60°C for 6 h to obtain a powder;

[0097] Step 2: Mix the powder obtained in Step 1 with pure water, yeast powder, and sodium dihydrogen phosphate to obtain a culture medium, and in the culture medium, the mass fraction of yeast powder is 2%, the mass fraction of sodium dihydrogen phosphate is 1%, the mass fraction of pure water is 17%, and the mass fraction of the powder is 80%;

[0098] Step 3: Inoculate the galactose yeast fermentation broth into the culture medium and ferment for 6 h, then centrifuge and take the supernatant to obtain a composition containing the fermentation product;

[0099] And the mass ratio of the galactose yeast fermentation broth in Step 3 to the powder in Step 1 is 87.5:5.

[0100] Comparative Example 2

[0101] Step 1: Dry and crush cordyceps sinensis at 60°C for 6 h to obtain a powder;

[0102] Step 2: Mix the powder obtained in Step 1 with pure water, yeast powder, and sodium dihydrogen phosphate to obtain a culture medium, and in the culture medium, the mass fraction of yeast powder is 2%, the mass fraction of sodium dihydrogen phosphate is 1%, the mass fraction of pure water is 17%, and the mass fraction of the powder is 80%;

[0103] Step 3: Inoculate the galactose yeast fermentation broth into the culture medium and ferment for 6 h, centrifuge to obtain the bottom liquid, then sterilize the bottom liquid at 121°C, crush it, and centrifuge again to obtain a composition containing the cell lysate;

[0104] And the mass ratio of the galactose yeast fermentation broth in Step 3 to the powder in Step 1 is 87.5:5.

[0105] Comparative Example 3

[0106] Basically the same as Example 1, except that cordyceps militaris of the same quality is used to replace cordyceps sinensis.

[0107] Comparative Example 4

[0108] It is basically the same as Example 1, except that a fermentation broth of Saccharomyces boulardii is prepared by using the same mass of Saccharomyces boulardii instead of Saccharomyces galactomyces.

[0109] Comparative Example 5

[0110] It is basically the same as Example 1, except that a fermentation broth of Saccharomyces cerevisiae is prepared by using the same mass of Saccharomyces cerevisiae instead of Saccharomyces galactomyces.

[0111] Comparative Example 6

[0112] It is basically the same as Example 1, except that the fruiting body of Cordyceps sinensis of the same mass is used to replace Cordyceps sinensis.

[0113] Comparative Example 7

[0114] It is basically the same as Example 1, except that the stroma of Cordyceps sinensis of the same mass is used to replace Cordyceps sinensis.

[0115] Performance test:

[0116] 1. Irritation test

[0117] 1.1 Instrument and equipment

[0118] Fully automatic incubator, stereomicroscope, SPF chicken embryos;

[0119] 1.2 Reagents

[0120] Sodium chloride, sodium dodecyl sulfate (SDS);

[0121] 1.3 Incubation conditions

[0122] Room temperature 20°C - 25°C, relative humidity 45% - 70%, incubation temperature 37.5°C ± 0.5°C, relative humidity 55% - 70%, turntable 3 times / h - 6 times / h, chicken embryos at 9 days old do not need to be rotated during incubation.

[0123] 1.4 Test method

[0124] (1) Test operation steps

[0125] In this test, 6 embryos are selected for each group, and the situation of the chorioallantoic membrane is recorded with a photographing device. A polytetrafluoroethylene resin ring is placed on the chorioallantoic membrane of the chicken embryo and photographed. The composite plant fermentation products of the examples and comparative examples are diluted with pure water to a solution with a mass fraction of 1% of the composite plant fermentation products and used as the test samples. Subsequently, the test samples are added into the polytetrafluoroethylene resin ring, the time of adding the samples is recorded, and the air chamber is covered with a moistened plastic wrap. The chicken embryos are transferred to an incubator with constant temperature and humidity for cultivation, and the degree of change in each toxic effect is observed.

[0126] (2) Result observation

[0127] Observe and record the manifestations of bleeding, blood coagulation and vascular lysis, and score according to their severity.

[0128] The scoring criteria for bleeding, blood coagulation and vascular lysis are shown in Table 1:

[0129] Table 1

[0130]

[0131] Scoring criteria: ES ≤ 12: no / mild irritation; 12 < ES < 16: moderate irritation; ES ≥ 16: strong irritation.

[0132] According to the test, neither the examples nor the comparative examples produced irritation. Refer to Figure 1-2 , for the comparison of the state of the chicken embryo before and after dropping the solution prepared from the composition of Example 1. After statistics, the ES value of the solution prepared from the composition of Example 1 was 4.0.

[0133] 2. Elastase inhibition rate test

[0134] 2.1 Instrument and equipment

[0135] BSA224S analytical balance, RT-6100 microplate reader;

[0136] 2.2 Reagents

[0137] Elastase (porcine pancreas), BR, N-succinyl-L-alanyl-L-alanyl-L-alanine, 98%;

[0138] 2.3 Test method

[0139] (1) Treatment of control and test samples

[0140] Sample group: The sample concentration is 100%;

[0141] Negative control: pure water.

[0142] (2) Test operation steps

[0143] Set up a sample group, a sample background group, a solvent group and a solvent background group. Each group needs to have 3 parallels. Add different reagent solutions to the 96-well plate, shake gently, incubate at 25 °C for 15 min, then place it in the microplate reader and measure the absorbance at 410 nm.

[0144] (3) The calculation formula is shown in Equation 1:

[0145] Equation 1;

[0146] Where: A—the absorbance of the reaction solution without the sample;

[0147] B—the absorbance of the reaction solution without the sample and enzyme;

[0148] C—the absorbance of the reaction solution containing the sample and enzyme;

[0149] D—the absorbance of the reaction solution containing the sample and without enzyme.

[0150] (4) Data analysis

[0151] The statistical analysis software is SPSS. The independent-samples t-test is used for the comparison of elastase inhibition rates among the test samples, positive control substances, and negative control substances. The above statistical analyses are all two-tailed tests, and the significance level is α = 0.05. P > 0.05 indicates no significant difference between the two groups; P < 0.05 indicates a significant difference between the two groups.

[0152] The test results are shown in Table 2:

[0153] Table 2

[0154] Item Elastase inhibition rate (%) P value Item Elastase inhibition rate (%) P value Example 1 42.574 ＜0.05 Example 11 40.987 ＜0.05 Example 2 42.358 ＜0.05 Example 12 41.031 ＜0.05 Example 3 42.101 ＜0.05 Example 13 40.846 ＜0.05 Example 4 42.387 ＜0.05 Control Example 1 35.715 ＜0.05 Example 5 43.971 ＜0.05 Control Example 2 36.484 ＜0.05 Example 6 44.256 ＜0.05 Control Example 3 38.154 ＜0.05 Example 7 44.171 ＜0.05 Control Example 4 35.152 ＜0.05 Example 8 44.109 ＜0.05 Control Example 5 36.106 ＜0.05 Example 9 45.876 ＜0.05 Control Example 6 35.182 ＜0.05 Example 10 40.449 ＜0.05 Control Example 7 34.569 ＜0.05 Negative control -3.415 /

[0155] Result analysis:

[0156] 1. It can be seen from Examples 1-4 that when the mass of the galactose yeast fermentation broth and the powder in the medium is slightly adjusted, although the elastase inhibition rate of the compositions prepared in Examples 1-4 fluctuates to a certain extent, the fluctuation range is small. It can be seen that when the mass of the galactose yeast and the powder in the medium is slightly adjusted, the fluctuation of the content of the substance with elastase inhibition ability in the composition is small;

[0157] 2. As can be seen from Example 1 and Examples 5 - 9, when the powder is replaced with a mixture of Cordyceps sinensis and Tremella fuciformis, the inhibitory ability of elastase in Example 5 has been significantly improved compared to Example 1. When mixtures of wheat, barley, buckwheat, rye, etc. are further incorporated into the powder in Examples 6, 7, and 8, the elastase inhibition rates of Examples 6, 7, and 8 are further improved compared to Example 5, but the improvement is relatively small. When the mixed grains of wheat, rye, and buckwheat are used in Example 9, the elastase inhibition rate of Example 9 is significantly improved compared to Examples 5 - 8. However, it is observed that the elastase inhibition rates of Examples 6 and 8 do not show a significant improvement compared to Example 5. After combining the grains used in Examples 6 and 8, Example 9 achieves an elastase inhibitory ability far exceeding that of Examples 6 and 8. It is speculated that the reason for this phenomenon may be that the mixed grains in Example 9 provide rich carbohydrates, proteins, fats, vitamins, and minerals and accelerate the growth and metabolism of yeast;

[0158] 3. As can be seen from Example 1 and Example 10, even when the galactose yeast of the present application is replaced with a commercially available galactose yeast, the inhibitory ability of elastase in Example 10 still shows an obvious downward trend. It can be seen that even though they belong to the same type of galactose yeast, the commercially available galactose yeast still cannot achieve an effect similar to that of the galactose yeast ( Geotrichum candidum ); Geotrichum candidum ).

[0159] Further observation of Examples 11 - 13 shows that when Tremella fuciformis is replaced with other plants rich in polysaccharide substances, Examples 11 - 13 cannot obtain results similar to those of Example 5;

[0160] 4. As can be seen from Example 1 and Comparative Examples 1 and 2, when the composition contains only the fermentation product or the lysate, the inhibitory ability of elastase in the composition shows an obvious downward trend. It can be seen that a single fermentation product or lysate cannot provide sufficient active ingredients with elastase inhibitory ability;

[0161] 5. As can be seen from Example 1 and Comparative Examples 3 - 5, when Cordyceps militaris similar to Cordyceps sinensis is used to replace Cordyceps sinensis or other types of yeast are used to replace galactose yeast, the elastase inhibition rates of Comparative Examples 3 - 5 show an obvious downward trend compared to Example 1. It can be seen that the use of Cordyceps sinensis and galactose yeast in the present application is irreplaceable;

[0162] Further observations on Comparative Examples 6-7 show that when only the fruiting bodies of Cordyceps sinensis and the worm-shaped sclerotia of Cordyceps sinensis are used in Comparative Examples 6 and 7 respectively, the elastase inhibition rates of Comparative Examples 6 and 7 both show an obvious downward trend. It can be seen that when the fruiting bodies of Cordyceps sinensis and the worm-shaped sclerotia of Cordyceps sinensis are used alone, they cannot provide enough nutrients for the decomposition by galactose yeast.

[0163] 3. Tyrosinase inhibition rate test

[0164] 3.1 Instrument and equipment

[0165] BSA224S analytical balance;

[0166] RT-6100 microplate reader;

[0167] 3.2 Reagents

[0168] Polyphenol oxidase (mushroom), BR;

[0169] L-DOPA, BR;

[0170] 3.3 Test method

[0171] (1) Treatment of control and test samples

[0172] Sample group: The sample concentration is 100%;

[0173] (2) Test operation steps

[0174] Set up sample tubes, sample background tubes, enzyme reaction tubes and solvent background tubes. Three parallel tubes need to be set up for each group. Add different reagent solutions to the four groups respectively, shake gently, and let stand at room temperature for 5 minutes. Transfer the reaction solutions of each group into 1 cm cuvettes and measure the absorbance at 475 nm.

[0175] (3) The calculation formula is shown in Equation 2:

[0176] Equation 2;

[0177] Where: T—the absorbance of the sample tube, that is, the absorbance of the solution after the reaction of the sample and tyrosinase;

[0178] T 0 —the absorbance of the sample background;

[0179] C—the average value of the absorbance of the enzyme reaction tube three times, that is, the absorbance of the reaction of tyrosinase and dopa without adding the sample;

[0180] C 0 —the absorbance of the solvent background.

[0181] (4) Data analysis

[0182] The statistical analysis software was SPSS. The independent samples t-test was used to compare the tyrosinase inhibition rates among the test samples, positive control substances, and negative control substances. All the above statistical analyses were two-tailed tests, and the significance level was α = 0.05. P > 0.05 indicates no significant difference between the two groups; P < 0.05 indicates a significant difference between the two groups.

[0183] The test results are shown in Table 3:

[0184] Table 3

[0185] Item Tyrosinase inhibition rate (%) P value Item Tyrosinase inhibition rate (%) P value Example 1 90.305 ＜0.05 Example 11 88.513 ＜0.05 Example 2 89.975 ＜0.05 Example 12 88.157 ＜0.05 Example 3 90.103 ＜0.05 Example 13 88.303 ＜0.05 Example 4 90.135 ＜0.05 Control Example 1 80.103 ＜0.05 Example 5 90.608 ＜0.05 Control Example 2 80.034 ＜0.05 Example 6 90.630 ＜0.05 Control Example 3 79.835 ＜0.05 Example 7 90.976 ＜0.05 Control Example 4 81.066 ＜0.05 Example 8 90.706 ＜0.05 Control Example 5 80.360 ＜0.05 Example 9 91.864 ＜0.05 Control Example 6 79.626 ＜0.05 Example 10 87.204 ＜0.05 Control Example 7 80.369 ＜0.05 Negative control -2.182 /

[0186] Result analysis:

[0187] 1. As can be seen from Examples 1-4, when the mass of galactose yeast and the powder in the culture medium was slightly adjusted, although the tyrosinase inhibition rate of the compositions prepared in Examples 1-4 fluctuated to a certain extent, the fluctuation range was small. It can be seen that when the mass of galactose yeast and the powder in the culture medium was slightly adjusted, the content of the substance with tyrosinase inhibition ability in the composition fluctuated little;

[0188] 2. As can be seen from Examples 1 and 5-9, when the powder was replaced with a mixture of Cordyceps sinensis and Tremella fuciformis, and when a mixture of wheat, barley, and buckwheat or a mixture of rye and buckwheat was further mixed into the powder in Examples 6, 7, and 8, the tyrosinase inhibition rate of Examples 6, 7, and 8 increased to a certain extent compared with Example 1, but the increase amplitude was small; when Example 9 used a mixed grain of wheat, rye, and buckwheat, the tyrosinase inhibition rate of Example 9 increased significantly compared with Examples 6-8. However, it was observed that the tyrosinase inhibition rate of Examples 6 and 8 did not increase significantly compared with Example 5, and Example 9 achieved a tyrosinase inhibition ability far exceeding that of Examples 5, 6, and 8 after combining the grains and Tremella fuciformis used in Examples 6 and 8. It is speculated that the reason for this phenomenon may be that the mixed grain in Example 9 provided rich carbohydrates, proteins, fats, vitamins, polysaccharides, and minerals and accelerated the growth and metabolism of yeast;

[0189] 3. As can be seen from Examples 1 and 10, even when galactose yeast was used to replace the galactose yeast of the present application ( Geotrichum candidum ), the tyrosinase inhibition ability of Example 10 still showed an obvious downward trend. It can be seen that even though they are both galactose yeast, the commercially available galactose yeast still cannot achieve an effect similar to that of galactose yeast ( Geotrichum candidum );

[0190] Further observation of Examples 11-13 shows that when other plants rich in polysaccharides are used to replace Tremella fuciformis, Examples 11-13 cannot achieve results similar to those of Example 5;

[0191] 4. It can be seen from Example 1 and Comparative Examples 1 and 2 that when the composition contains only fermentation products or lysates, the tyrosinase inhibitory ability of the composition has a significant downward trend. It can be seen that a single fermentation product or lysate cannot provide enough active ingredients with tyrosinase inhibitory ability;

[0192] 5. It can be seen from Example 1 and Comparative Examples 3-5 that when Cordyceps militaris, which is similar to Cordyceps sinensis, is used to replace Cordyceps sinensis or other types of yeast are used to replace Galacto-yeast, the tyrosinase inhibition rates of Comparative Examples 3-5 relative to Example 1 all show a significant downward trend. It can be seen that the use of Cordyceps sinensis and Galacto-yeast in this application is irreplaceable;

[0193] Further observation of Comparative Examples 6-7 shows that when Comparative Examples 6 and 7 respectively use only the fruiting body of Cordyceps sinensis and the insect-shaped sclerotium of Cordyceps sinensis, the tyrosinase inhibition rates of Comparative Examples 6 and 7 both have a significant downward trend. It can be seen that the fruiting body of Cordyceps sinensis and the insect-shaped sclerotium of Cordyceps sinensis cannot provide sufficient nutrients for galactose yeast to decompose when used alone.

[0194] 4. Peptide identification and analysis test

[0195] The amino acids of the small molecule peptide composition obtained by fermenting Cordyceps sinensis obtained in Example 1 were identified and analyzed using a high-resolution liquid spectrometer.

[0196] 4.1 Materials and instruments

[0197] Mass spectrometry grade acetonitrile and formic acid were from Sigma, USA; ultrapure water was homemade using a Milli-Q water purifier (Millipore, Germany).

[0198] 100,000-bit analytical electronic balance, Sartorius, Germany; high-resolution liquid chromatography-mass spectrometry (X500LC-ESI-Q-TOF, AB SCIEX, USA); liquid chromatography and mass spectrometry control software using SCIEX OS 2.0 integrated version (AB SCIEX, USA); chromatographic column using 1×100mm HSS T3 (1.8μm, 100 A, Waters, USA).

[0199] 4.2 Sample preparation

[0200] The liquid sample was diluted to prepare a 2 mg / mL solution, and then passed through a 0.22 μm organic filter membrane and analyzed on an analyzer.

[0201] 4.3 Detection methods

[0202] The samples were separated and detected using an X500 LC-ESI-Q-TOF high-resolution liquid chromatography-mass spectrometry (AB Sciex, USA). The mobile phase consisted of 0.1% (v / v) formic acid aqueous solution (A) and acetonitrile (B). The elution method was as follows: 5.0% B from 0 - 4.00 min, 5.0 - 10.0% B from 4.00 - 6.00 min, 10.0 - 40.0% B from 6.00 - 30.00 min, 40.0 - 90.0% B from 30.00 - 34.00 min, 90% B from 34.00 - 40.00 min, 90.0 - 5.0% B from 40.00 - 42.00 min, 5.0% B from 42.00 - 52.00 min, the flow rate was 0.05 mL / min, the injection volume was 1 μL, and the column temperature was 40 °C. The mass spectrometry detection method: the scan cycle was 0.642 s, the ESI ion source temperature was 500 °C, in positive ion mode, the spray voltage was 5500 V, the TOF primary scan range was 100 - 1200 Da, the secondary scan range was 50 - 1200 Da, the working mode was IDA, the maximum number of candidate ions was 4, dynamic exclusion was enabled, and the remaining parameters used the default values of the proteomics method. Note that before using the instrument, its dead volume should be minimized.

[0203] 4.4 Data analysis

[0204] Data format conversion (wiff2 to Mgf and mzMXL) was performed using the open-source software ProteoWizard 3.0. Polypeptide group identification and analysis were performed using PspOS 2.0.1 (Wuyi University, Jiangmen, Guangdong), with the dual-engine setting enabled. The identification length was set to 2 - 30, where the infinite search engine length was set to 2 - 6, and the sequence library search identification length was 7 - 30 (the sequence library was downloaded from Uniprot: Viridiplantae-reviewed, 2023-11-11). The primary parent ion error was 20 ppm, the secondary daughter ion error was 0.02 Da, the detection rate threshold for mixed ion clusters was 50%, the Bayesian score threshold was 36, the maximum drift time of the chromatographic peak was 0.50 min, and the number of parallel deconvolution cores was 6.

[0205] The test results showed that the chromatogram of the small molecule peptide composition prepared by fermenting Cordyceps sinensis in Example 1 was as shown in Figure 3 shown, and the mass spectrometry diagram was as shown in Figure 4 shown. The structure diagram and structure analysis of the bioactive polypeptide were as shown in Figure 5 shown.

[0206] Application examples

[0207] A small molecule peptide composition essence prepared by fermenting Cordyceps sinensis, and the specific formula is shown in Table 4 as follows:

[0208] Table 4

[0209] Raw material name Addition amount wt% Small molecule peptide composition prepared by fermenting Cordyceps sinensis in Example 1 20 Glycerol 2 Carbomer 0.12 Xanthan gum 0.15 Sodium hyaluronate 0.1 Arginine 0.12 1,2-Hexanediol 0.5 1,3-Butanediol 7 p-Hydroxyacetophenone 0.5 Water Add up to 100%

[0210] The above embodiments are preferred embodiments of the present invention. However, the embodiments of the present invention are not limited by the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications made without departing from the spirit and principle of the present invention shall be equivalent replacement methods and are all included in the protection scope of the present invention.

Claims

1. A small molecule peptide composition obtained by fermenting Cordyceps sinensis, characterized in that: The preparation method of the small molecule peptide composition obtained by fermentation of Cordyceps sinensis is specifically as follows: Step 1: Dry and crush cordyceps sinensis, tremella fuciformis and grains to obtain powder, wherein the mass ratio of cordyceps sinensis, tremella fuciformis and grains is 5:1:1; Step 2: Mix the powder obtained in step 1 with pure water, yeast powder and sodium dihydrogen phosphate to obtain a culture medium, wherein the mass fraction of yeast powder in the culture medium is 2%, the mass fraction of sodium dihydrogen phosphate is 1%, the mass fraction of pure water is 92%, and the mass fraction of the powder is 5%; Step 3: inoculating the galactose yeast fermentation liquid into the culture medium and fermenting for 6 hours, then sterilizing at 121° C., crushing, centrifuging, and taking the supernatant to obtain a small molecule peptide composition obtained by fermentation of Cordyceps sinensis containing fermentation products and lysate; The cereals are a mixture of wheat, rye and buckwheat, and the mass ratio of wheat, rye and buckwheat is 2:1.5:3; The mass ratio of the galactose yeast fermentation liquid in step 3 to the powder in step 1 is 87.5:

5.

2. Use of the small molecule peptide composition obtained by fermentation of Cordyceps sinensis as claimed in claim 1 for preparing cosmetics.

3. A cosmetic, characterized in that: The invention relates to a small molecule peptide composition containing 0.1 to 100 wt % of the small molecule peptide composition obtained by fermenting Cordyceps sinensis as claimed in claim 1.

4. The cosmetic according to claim 3, characterized in that: The invention also comprises a solvent, wherein the solvent is selected from at least one of glycerol and butylene glycol.

5. The cosmetic according to claim 3, characterized in that: The dosage form of the cosmetic is water solution, essence, facial mask liquid, emulsion or cream.

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