A yeast fermentation product with penetration-promoting efficacy, and a preparation method and application thereof

Abstract

The present application provides a yeast fermentation product with penetration promoting effect, and a preparation method and application thereof. First, Saccharomyces boulardii and Schizosaccharomyces pombe are inoculated in a fermentation medium with specific components at a mass ratio of 0.7-1.3:0.7-1.3, and then micro-aerobic fermentation is carried out at a temperature of 25-35 DEG C and an oxygen content of less than or equal to 10% for 48-72 hours, followed by inoculation of Saccharomyces cerevisiae, and then micro-aerobic fermentation is carried out at a temperature of 26-32 DEG C and an oxygen content of less than or equal to 5% for 48-72 hours. The yeast fermentation product obtained by mixing and fermenting Saccharomyces boulardii, Schizosaccharomyces pombe and Saccharomyces cerevisiae in a specific order and in a specific medium has a more excellent penetration promoting effect than products obtained by other fermentation methods, can significantly promote the transdermal absorption of active ingredients in cosmetic or medical compositions, and is suitable for preparing cosmetic or medical compositions.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of microbial fermentation. More particularly, it relates to a yeast fermentation product with penetration-promoting effect, and a preparation method and application thereof. BACKGROUND

[0002] Skin is the barrier between the human body and the external environment, playing an important role in helping to block harmful substances from the outside world and preventing the loss of water or nutrients. However, this strong barrier function also hinders the absorption of active ingredients in cosmetics into the skin. Therefore, in order to overcome the barrier function of the skin and promote the penetration of active ingredients into the deep layer of the skin, thereby exerting various physiological regulatory effects on skin cells, it is often necessary to add certain penetration enhancers to improve the transdermal absorption of active ingredients in the preparation of cosmetics.

[0003] However, existing chemical penetration enhancers, such as azone, surfactants, organic acids, etc., all have certain irritant properties and can easily cause skin allergies, and are not suitable for long-term use. Therefore, researchers have gradually shifted their focus to microbial fermentation products that are safe and friendly in composition. Microorganisms can produce secondary metabolites such as biological enzymes during fermentation, which can decompose biological macromolecules such as proteins and polysaccharides into smaller molecules, thereby playing a penetration-promoting role. However, the fermentation products obtained by different microorganisms and different fermentation methods have significant differences, and the penetration-promoting effect varies greatly. SUMMARY

[0004] The present application aims to provide a preparation method of a yeast fermentation product, which is obtained by mixed fermentation of Brettanomyces bruxellensis, Schizosaccharomyces pombe and Saccharomyces cerevisiae in a specific order and in a specific culture medium. Compared with products obtained by other fermentation methods, the yeast fermentation product has more excellent penetration-promoting effect, can significantly promote the transdermal absorption of active ingredients in cosmetic or medical compositions, and is suitable for preparing cosmetic or medical compositions.

[0005] The first object of the present application is to provide a preparation method of a yeast fermentation product.

[0006] The second object of the present application is to provide a yeast fermentation product prepared by the above method.

[0007] The third object of the present application is to provide the use of the above yeast fermentation product in the preparation of cosmetic or medical compositions.

[0008] The fourth object of the present application is to provide a cosmetic or medical composition.

[0009] The above objects of the present application are achieved by the following technical solutions:

[0010] The application provides a preparation method of a yeast fermentation product, comprising the following steps:

[0011] S1. inoculating the Brettanomyces bruxellensis and the Schizosaccharomyces pombe in the fermentation medium at a mass ratio of 0.7-1.3:0.7-1.3, and carrying out micro-aerobic fermentation at a temperature of 25-35 DEG C and an oxygen content of less than or equal to 10% for 48-72 hours;

[0012] S2. inoculating the Saccharomyces cerevisiae in the product obtained in S1, and carrying out micro-aerobic fermentation at a temperature of 26-32 DEG C and an oxygen content of less than or equal to 5% for 48-72 hours to obtain the product;

[0013] In the method, the fermentation medium in S1 has a pH of 6-7 and comprises the following components in the following mass fractions: 0.8-1.2 parts of mint, 1-3 parts of rice, 0.8-1.2 parts of soybean powder, 0.8-1.2 parts of Danshen, 0.2-0.4 parts of NaCl, 0.1-0.2 parts of K2HPO4 and 60-100 parts of water.

[0014] Preferably, the mass ratio of the Brettanomyces bruxellensis to the Schizosaccharomyces pombe in S1 is 1:1.

[0015] Preferably, the fermentation medium in S1 has a pH of 6-7 and comprises the following components in the following mass fractions: 1 part of mint, 2 parts of rice, 1 part of soybean powder, 1 part of Danshen, 0.3 parts of NaCl, 0.15 parts of K2HPO4 and 80 parts of water.

[0016] Preferably, the fermentation medium is sterilized before inoculation in S1.

[0017] Further preferably, the sterilization is carried out at 115-125 DEG C for 15-25 minutes, and most preferably, the sterilization is carried out at 121 DEG C for 20 minutes.

[0018] Preferably, the total inoculation amount of the Brettanomyces bruxellensis and the Schizosaccharomyces pombe in the fermentation medium in S1 is 3wt%-5wt%.

[0019] The Brettanomyces bruxellensis and the Schizosaccharomyces pombe are activated by a conventional method before inoculation, for example, the Brettanomyces bruxellensis is cultured in YPDA medium to a cell density of 1x10 7 -1x10 8 cfu / mL, and the Schizosaccharomyces pombe is cultured in CM0181 medium to a cell density of 1x10 7 -1x10 8 cfu / mL.

[0020] Preferably, the micro-aerobic fermentation in S1 is carried out at 180-250 rpm.

[0021] Preferably, in S2, the inoculation amount of the Saccharomyces cerevisiae in the product obtained in S1 is 4wt%-10wt%.

[0022] The Saccharomyces cerevisiae is activated in a conventional way before inoculation, for example, cultured in PDB medium to a cell density of 1×10 7 ~1×10 8 cfu / mL.

[0023] Preferably, the micro-aerobic fermentation in S2 is carried out at 100-300rpm.

[0024] Preferably, after the micro-aerobic fermentation in S2, ultrasonic treatment and solid-liquid separation are further carried out.

[0025] Further preferably, the ultrasonic treatment is carried out at 0.05-0.15MPa for 1.8-2.2h.

[0026] More preferably, the ultrasonic treatment is carried out simultaneously with stirring, for example, at 100-200rpm.

[0027] Further preferably, the solid-liquid separation is filtration.

[0028] More preferably, the filtration is carried out to obtain a filtrate with a particle size of ≤10μm.

[0029] Further preferably, after the solid-liquid separation, sterilization is further carried out.

[0030] More preferably, the sterilization is carried out at 80-90℃ for 20-30min.

[0031] More preferably, after the sterilization, cooling is further carried out, for example, to a temperature below 45℃.

[0032] The yeast fermentation product prepared by the above method has a high content of small molecular polypeptides and amino acids, and has a strong transdermal penetration effect, which can significantly promote the transdermal absorption of active ingredients in a cosmetic or medical composition, and better exert the original effect of the cosmetic or medical composition. Therefore, the yeast fermentation product prepared by the above method, the use of the yeast fermentation product in the preparation of a cosmetic or medical composition, and a cosmetic or medical composition comprising the yeast fermentation product and a cosmetically or medically acceptable adjuvant should all be within the protection scope of the present application.

[0033] The present application has the following beneficial effects:

[0034] The present application is prepared by mixed fermentation of Brettanomyces bruxellensis, Schizosaccharomyces pombe and Saccharomyces cerevisiae in a specific order and in a specific medium, and has more excellent penetration efficacy than products obtained by other fermentation methods, and can significantly promote the transdermal absorption of active ingredients in cosmetic or medical compositions, and is suitable for preparing cosmetic or medical compositions. DETAILED DESCRIPTION

[0035] The present application is further illustrated below in conjunction with specific examples, but the examples do not limit the present application in any form. Unless otherwise specified, the reagents, methods and equipment used in the present application are conventional reagents, methods and equipment in the technical field.

[0036] Unless otherwise specified, the reagents and materials used in the following examples are commercially available.

[0037] Activation of Brettanomyces bruxellensis: cultured in YPDA medium to a cell density of 1×10 8 cfu / mL.

[0038] Activation of Schizosaccharomyces pombe: cultured in CM0181 medium to a cell density of 1×10 8 cfu / mL.

[0039] Activation of Saccharomyces cerevisiae: cultured in PDB medium to a cell density of 1×10 8 cfu / mL.

[0040] Example 1: A method for preparing a yeast fermentation product

[0041] S1. Sterilize the fermentation medium at 121℃ for 20 min, then inoculate Brettanomyces bruxellensis and Schizosaccharomyces pombe into the fermentation medium at a mass ratio of 1:1 (so that the total inoculation amount of Brettanomyces bruxellensis and Schizosaccharomyces pombe in the fermentation medium is 4wt%), and perform micro-aerobic fermentation at a temperature of 30℃, an oxygen content of 8%, and a rotation speed of 220rpm for 60h in a constant-temperature shaker;

[0042] S2. Inoculate Saccharomyces cerevisiae into the product obtained in S1 (so that the inoculation amount of Saccharomyces cerevisiae in the product obtained in S1 is 7wt%), and perform micro-aerobic fermentation at a temperature of 29℃, an oxygen content of 3%, and a rotation speed of 200rpm for 60h in a constant-temperature shaker, then perform ultrasonic treatment at 0.1MPa and 150rpm for 2h, and filter with a 10μm filter membrane, and then sterilize the obtained filtrate at 85℃ for 25min, and then cool it to 45℃;

[0043] The fermentation medium in S1 has a pH of 6.5, and contains the following components by mass: mint 1 part, rice 2 parts, soybean powder 1 part, Danshen 1 part, NaCl 0.3 part, K2HPO4 0.15 part, and water 80 parts.

[0044] Example 2 A method for preparing a yeast fermentation product

[0045] S1. The fermentation culture was sterilized at 125℃ for 15 min, and then the Brettanomyces bruxellensis and the Schizosaccharomyces pombe were inoculated into the fermentation medium at a mass ratio of 1.3:0.7 (so that the total inoculation amount of the Brettanomyces bruxellensis and the Schizosaccharomyces pombe in the fermentation medium was 3 wt%), and subjected to micro-aerobic fermentation at a temperature of 35℃, an oxygen content of 10%, and a rotation speed of 180 rpm for 48 h in a constant-temperature shaker;

[0046] S2. The Saccharomyces cerevisiae was inoculated into the product obtained in S1 (so that the inoculation amount of the Saccharomyces cerevisiae in the product obtained in S1 was 10 wt%), and subjected to micro-aerobic fermentation at a temperature of 26℃, an oxygen content of 5%, and a rotation speed of 300 rpm for 48 h in a constant-temperature shaker, and then subjected to ultrasonic treatment at 0.05 MPa and a rotation speed of 200 rpm for 1.8 h, and filtered with a 10 μm filter membrane, and the obtained filtrate was sterilized at 80℃ for 30 min, and then cooled to 45℃.

[0047] In the fermentation medium of S1, the pH was 7, and the fermentation medium contained the following components in the following amounts by mass: mint 0.8 parts, rice 3 parts, soybean powder 0.8 parts, Danshen 0.8 parts, NaCl 0.4 parts, K2HPO4 0.1 parts, and water 60 parts.

[0048] Example 3 A method for preparing a yeast fermentation product

[0049] S1. The fermentation culture was sterilized at 115℃ for 25 min, and then the Brettanomyces bruxellensis and the Schizosaccharomyces pombe were inoculated into the fermentation medium at a mass ratio of 0.7:1.3 (so that the total inoculation amount of the Brettanomyces bruxellensis and the Schizosaccharomyces pombe in the fermentation medium was 5 wt%), and subjected to micro-aerobic fermentation at a temperature of 25℃, an oxygen content of 5%, and a rotation speed of 250 rpm for 72 h in a constant-temperature shaker;

[0050] S2. The Saccharomyces cerevisiae was inoculated into the product obtained in S1 (so that the inoculation amount of the Saccharomyces cerevisiae in the product obtained in S1 was 4 wt%), and subjected to micro-aerobic fermentation at a temperature of 32℃, an oxygen content of 2%, and a rotation speed of 100 rpm for 72 h in a constant-temperature shaker, and then subjected to ultrasonic treatment at 0.15 MPa and a rotation speed of 100 rpm for 2.2 h, and filtered with a 10 μm filter membrane, and the obtained filtrate was sterilized at 90℃ for 20 min, and then cooled to 45℃.

[0051] In the fermentation medium of S1, the pH was 6, and the fermentation medium contained the following components in the following amounts by mass: mint 1.2 parts, rice 1 part, soybean powder 1.2 parts, Danshen 1.2 parts, NaCl 0.2 parts, K2HPO4 0.2 parts, and water 100 parts.

[0052] Comparative Example 1

[0053] The same as example 1, except that the Saccharomyces cerevisiae is inoculated first, and then the Brettanomyces bruxellensis and the Schizosaccharomyces pombe are inoculated.

[0054] That is, the present comparative example is specifically as follows:

[0055] S1. The fermentation culture is sterilized at 121℃ for 20 min, and then the Saccharomyces cerevisiae is inoculated into the fermentation culture (so that the inoculation amount of the Saccharomyces cerevisiae in the fermentation culture is 7wt%), and micro-aerobic fermentation is carried out at a temperature of 29℃, an oxygen content of 3%, and a constant-temperature shaker speed of 200rpm for 60h;

[0056] S2. The Brettanomyces bruxellensis and the Schizosaccharomyces pombe are inoculated into the product obtained in S1 at a mass ratio of 1:1 (so that the total inoculation amount of the Brettanomyces bruxellensis and the Schizosaccharomyces pombe in the product obtained in S1 is 4wt%), and micro-aerobic fermentation is carried out at a temperature of 30℃, an oxygen content of 8%, and a constant-temperature shaker speed of 220rpm for 60h, followed by ultrasonic treatment at 0.1MPa and 150rpm for 2h, and then filtration with a 10μm filter membrane, and the obtained filtrate is sterilized at 85℃ for 25min, and then cooled to 45℃;

[0057] The fermentation culture in S1 has a pH of 6.5, and contains the following components in mass parts: mint 1 part, rice 2 parts, soybean powder 1 part, Danshen 1 part, NaCl 0.3 part, K2HPO4 0.15 part, and water 80 parts.

[0058] Comparative example 2

[0059] The same as example 1, except that the Brettanomyces bruxellensis is replaced by Candida bombicola. That is, the present comparative example is specifically as follows:

[0060] S1. The fermentation culture is sterilized at 121℃ for 20 min, and then the Candida bombicola and the Schizosaccharomyces pombe are inoculated into the fermentation culture at a mass ratio of 1:1 (so that the total inoculation amount of the Candida bombicola and the Schizosaccharomyces pombe in the fermentation culture is 4wt%), and micro-aerobic fermentation is carried out at a temperature of 30℃, an oxygen content of 8%, and a constant-temperature shaker speed of 220rpm for 60h;

[0061] S2. The Saccharomyces cerevisiae is inoculated into the product obtained in S1 (so that the inoculation amount of the Saccharomyces cerevisiae in the product obtained in S1 is 7wt%), and micro-aerobic fermentation is carried out at a temperature of 29℃, an oxygen content of 3%, and a constant-temperature shaker speed of 200rpm for 60h, followed by ultrasonic treatment at 0.1MPa and 150rpm for 2h, and then filtration with a 10μm filter membrane, and the obtained filtrate is sterilized at 85℃ for 25min, and then cooled to 45℃;

[0062] The fermentation medium in S1 has a pH of 6.5 and contains the following components in parts by mass: mint 1 part, rice 2 parts, soybean powder 1 part, Danshen 1 part, NaCl 0.3 part, K2HPO4 0.15 part, and water 80 parts.

[0063] Comparative Example 3

[0064] The same as in Example 1, except that the Torulopsis candida is replaced by the Saccharomyces cerevisiae. That is, the present comparative example is as follows:

[0065] S1. The fermentation medium is sterilized at 121°C for 20 min, and then the Saccharomyces cerevisiae and the Torulopsis candida are inoculated into the fermentation medium at a mass ratio of 1:1 (so that the total inoculation amount of the Saccharomyces cerevisiae and the Torulopsis candida in the fermentation medium is 4 wt %), and subjected to micro-aerobic fermentation at a temperature of 30°C, an oxygen content of 8%, and a rotation speed of 220 rpm for 60 h in a constant-temperature shaker;

[0066] S2. The Saccharomyces cerevisiae is inoculated into the product obtained in S1 (so that the inoculation amount of the Saccharomyces cerevisiae in the product obtained in S1 is 7 wt %), and subjected to micro-aerobic fermentation at a temperature of 29°C, an oxygen content of 3%, and a rotation speed of 200 rpm for 60 h in a constant-temperature shaker, followed by ultrasonic treatment at 0.1 MPa and 150 rpm for 2 h, and then filtered with a 10 μm filter membrane. The obtained filtrate is sterilized at 85°C for 25 min, and then cooled to 45°C.

[0067] The fermentation medium in S1 has a pH of 6.5 and contains the following components in parts by mass: mint 1 part, rice 2 parts, soybean powder 1 part, Danshen 1 part, NaCl 0.3 part, K2HPO4 0.15 part, and water 80 parts.

[0068] Comparative Example 4

[0069] The same as in Example 1, except that the Saccharomyces cerevisiae is replaced by the Hansenula anomala. That is, the present comparative example is as follows:

[0070] S1. The fermentation medium is sterilized at 121°C for 20 min, and then the Saccharomyces cerevisiae and the Torulopsis candida are inoculated into the fermentation medium at a mass ratio of 1:1 (so that the total inoculation amount of the Saccharomyces cerevisiae and the Torulopsis candida in the fermentation medium is 4 wt %), and subjected to micro-aerobic fermentation at a temperature of 30°C, an oxygen content of 8%, and a rotation speed of 220 rpm for 60 h in a constant-temperature shaker;

[0071] S2. The abnormal Hansenula was inoculated into the product obtained in S1 (so that the inoculation amount of the abnormal Hansenula in the product obtained in S1 was 7 wt%), and subjected to micro-aerobic fermentation at 29℃, 3% oxygen content, and 200 rpm for 60 h in a constant-temperature shaker, then subjected to ultrasonic treatment at 0.1 MPa and 150 rpm for 2 h, filtered with a 10 μm filter, and the obtained filtrate was sterilized at 85℃ for 25 min, then cooled to 45℃.

[0072] The fermentation medium in S1 had a pH of 6.5 and contained the following components in parts by mass: mint 1 part, rice 2 parts, soybean powder 1 part, Danshen 1 part, NaCl 0.3 part, K2HPO4 0.15 part, and water 80 parts.

[0073] Comparative Example 5

[0074] The same as in Example 1, except that only Saccharomyces boulardii was inoculated in S1. That is, the present comparative example was as follows:

[0075] S1. The fermentation medium was sterilized at 121℃ for 20 min, then Saccharomyces boulardii was inoculated into the fermentation medium (so that the inoculation amount of Saccharomyces boulardii in the fermentation medium was 4 wt%), and subjected to micro-aerobic fermentation at 30℃, 8% oxygen content, and 220 rpm for 60 h in a constant-temperature shaker;

[0076] S2. The abnormal Hansenula was inoculated into the product obtained in S1 (so that the inoculation amount of the abnormal Hansenula in the product obtained in S1 was 7 wt%), and subjected to micro-aerobic fermentation at 29℃, 3% oxygen content, and 200 rpm for 60 h in a constant-temperature shaker, then subjected to ultrasonic treatment at 0.1 MPa and 150 rpm for 2 h, filtered with a 10 μm filter, and the obtained filtrate was sterilized at 85℃ for 25 min, then cooled to 45℃.

[0077] The fermentation medium in S1 had a pH of 6.5 and contained the following components in parts by mass: mint 1 part, rice 2 parts, soybean powder 1 part, Danshen 1 part, NaCl 0.3 part, K2HPO4 0.15 part, and water 80 parts.

[0078] Comparative Example 6

[0079] The same as in Example 1, except that only Schizosaccharomyces pombe was inoculated in S1. That is, the present comparative example was as follows:

[0080] S1. The fermentation medium was sterilized at 121℃ for 20 min, then Schizosaccharomyces pombe was inoculated into the fermentation medium (so that the inoculation amount of Schizosaccharomyces pombe in the fermentation medium was 4 wt%), and subjected to micro-aerobic fermentation at 30℃, 8% oxygen content, and 220 rpm for 60 h in a constant-temperature shaker;

[0081] S2. Saccharomyces cerevisiae was inoculated into the product obtained in S1 (the inoculation amount of Saccharomyces cerevisiae in the product obtained in S1 was 7 wt%), and micro-aerobic fermentation was carried out at a temperature of 29°C, an oxygen content of 3%, and a constant-temperature shaker speed of 200 rpm for 60 h, followed by ultrasonic treatment at 0.1 MPa and 150 rpm for 2 h, and then the obtained filtrate was filtered with a 10 μm filter membrane, and after sterilization at 85°C for 25 min, the temperature was lowered to 45°C.

[0082] The fermentation medium in S1 had a pH of 6.5 and contained the following components in parts by mass: mint 1 part, rice 2 parts, soybean powder 1 part, Danshen 1 part, NaCl 0.3 part, K2HPO4 0.15 part, and water 80 parts.

[0083] Comparative Example 7

[0084] The same as in Example 1, except that the rice in the fermentation medium was replaced by millet. That is, the present comparative example was as follows:

[0085] S1. The fermentation medium was sterilized at 121°C for 20 min, and then Brettanomyces bruxellensis and Schizosaccharomyces pombe were inoculated into the fermentation medium at a mass ratio of 1:1 (so that the total inoculation amount of Brettanomyces bruxellensis and Schizosaccharomyces pombe in the fermentation medium was 4 wt%), and micro-aerobic fermentation was carried out at a temperature of 30°C, an oxygen content of 8%, and a constant-temperature shaker speed of 220 rpm for 60 h.

[0086] S2. Saccharomyces cerevisiae was inoculated into the product obtained in S1 (the inoculation amount of Saccharomyces cerevisiae in the product obtained in S1 was 7 wt%), and micro-aerobic fermentation was carried out at a temperature of 29°C, an oxygen content of 3%, and a constant-temperature shaker speed of 200 rpm for 60 h, followed by ultrasonic treatment at 0.1 MPa and 150 rpm for 2 h, and then the obtained filtrate was filtered with a 10 μm filter membrane, and after sterilization at 85°C for 25 min, the temperature was lowered to 45°C.

[0087] The fermentation medium in S1 had a pH of 6.5 and contained the following components in parts by mass: mint 1 part, millet 2 parts, soybean powder 1 part, Danshen 1 part, NaCl 0.3 part, K2HPO4 0.15 part, and water 80 parts.

[0088] Test Example 1: Penetration-promoting effect of yeast fermentation product

[0089] The back skin of a 2-month-old piglet was fixed between the supply pool and the receiving pool of a Franz vertical two-chamber permeation diffusion cell, 12 mL of normal saline was added to the supply pool, and the receiving pool was not filled with any liquid. After standing for 30 min, if no significant subcutaneous liquid titration or outflow appeared on the receiving pool side, it was proved that the skin barrier function was intact, otherwise it was determined that the skin barrier function was damaged. After confirming that the skin barrier function was intact, the sample to be tested (10 mL of the yeast fermentation product obtained in Examples 1 to 3 and Comparative Examples 1 to 7 was mixed with 0.1 g of vitamin A as the sample to be tested) was added to the receiving pool. Then the Franz vertical two-chamber permeation diffusion cell was placed in a 37°C constant temperature water bath, and at the 24th hour of the water bath, sampling was performed from the receiving pool, and the content of vitamin A therein was determined by HPLC chromatography (Vertex chromatographic column, eluent was acetonitrile, methanol, dichloromethane in a volume ratio of 3:3:4, eluent flow rate was 1.0 mL / min, column temperature was 25°C). The transdermal absorption rate of the sample to be tested was calculated according to the formula "transdermal absorption rate (%) = mass of vitamin A in the receiving pool / mass of vitamin A in the sample to be tested x 100%". The results are shown in Table 1.

[0090] Table 1

[0091] Transdermal absorption rate (%) Example 1 58.2 Example 2 55.7 Example 3 53.2 Comparative Example 1 42.0 Comparative Example 2 32.1 Comparative Example 3 28.0 Comparative Example 4 29.4 Comparative Example 5 21.2 Comparative Example 6 19.5 Comparative Example 7 14.2

[0092] It can be seen that the transdermal absorption rate of the yeast fermentation product obtained in Examples 1 to 3 is significantly higher than that of Comparative Examples 1 to 7, indicating that the yeast fermentation product obtained by mixing and fermenting the Bruxellomyces, Schizosaccharomyces pombe and Saccharomyces cerevisiae in a specific order in a specific medium has more excellent penetration promoting effect compared with the product obtained by other fermentation methods, can significantly promote the transdermal absorption of active ingredients in cosmetic or medical compositions, and is suitable for preparing cosmetic or medical compositions.

[0093] Test Example 2 Transdermal ability of serum containing yeast fermentation product

[0094] I. Preparation of serum containing yeast fermentation product

[0095] (1) Ingredients

[0096] EDTA-2Na 0.05wt%, glycerin 1.00wt%, 1,3-butanediol 1.00wt%, sclerotium gum 0.12wt%, carbomer U20 0.17wt%, vitamin B5 2.00wt%, p-hydroxyacetophenone 0.60wt%, KOH 0.05wt%, 1,2-hexanediol 0.60wt%, squalane 1.50wt%, biosaccharide gum-1 1.00wt%, cyclomethicone 2.00wt%, hydroxyethyl acrylate / acryloyldimethyltaurate copolymer 0.35wt%, the yeast fermentation product of Examples 1 to 3 and Comparative Examples 1 to 7 3.00wt%, and water to 100wt%.

[0097] (2) Preparation method

[0098] The ingredients were mixed at 80°C to obtain the essence containing the yeast fermentation product.

[0099] II. Preparation of a control essence

[0100] (1) Ingredients

[0101] EDTA-2Na 0.05wt%, glycerin 1.00wt%, 1,3-butanediol 1.00wt%, sclerotium gum 0.12wt%, U21 0.17wt%, VB5 2.00wt%, p-hydroxyacetophenone 0.60wt%, KOH 0.05wt%, 1,2-hexanediol 0.60wt%, squalane 1.50wt%, biosaccharide gum-1 1.00wt%, cyclomethicone 2.00wt%, hydroxyethyl acrylate / acryloyldimethyltaurate copolymer 0.35wt%, and water to 100wt%.

[0102] (2) Preparation method

[0103] The ingredients were mixed at 80°C to obtain the essence containing the yeast fermentation product.

[0104] III. DPPH radical scavenging experiment

[0105] Sample group: The essence was dissolved in distilled water (to give a final concentration of 1% (v / v) of the essence), 100 μL was taken and added to a 96-well plate, 100 μL of a 0.1 mM DPPH solution was added, and the mixture was shaken in the dark for 10 min, and then the absorbance Ax at 520 nm was measured using an enzyme marker. Three duplicate wells were set up, and the results were averaged.

[0106] Control group: 100 μL distilled water was added to a 96-well plate, 100 μL of 0.1 mM DPPH solution was added, and the mixture was shaken in the dark for 10 min. The absorbance A0 at 520 nm was measured using an enzyme marker. Three duplicate wells were set up, and the results were averaged.

[0107] The DPPH radical scavenging rate of the serum was calculated according to the formula "DPPH radical scavenging rate (%) = [(A0-Ax) / A0]x100%". The results are shown in Table 2.

[0108] Table 2

[0109]

[0110]

[0111] It can be seen that the DPPH radical scavenging rate of the serum containing the yeast fermentation product obtained in Examples 1-3 is significantly higher than that of the serum containing the yeast fermentation product obtained in Comparative Examples 1-7 and the control serum, indicating that the yeast fermentation product obtained by mixing and fermenting B. bruxellensis, S. pombe, and S. cerevisiae in a specific order in a specific medium has a more excellent penetration-promoting effect than other fermentation products, can significantly promote the transdermal absorption of active ingredients in cosmetic or medical compositions, and is suitable for preparing cosmetic or medical compositions.

[0112] The above examples are preferred embodiments of the present application, but the embodiments of the present application are not limited to the above examples, and any changes, modifications, substitutions, combinations, simplifications made without departing from the spirit and principles of the present application are equivalent replacement methods and are included in the protection scope of the present application.

Claims

1. A method for preparing yeast fermentation products, characterized in that, Includes the following steps: S1. Inoculate Brussels wine yeast and chestnut wine schizosacchari at a mass ratio of 0.7-1.3:0.7-1.3 into the fermentation medium and ferment microaerobicly at a temperature of 25-35 ℃ and an oxygen content of ≤10% for 48-72 h. S2. Inoculate the product obtained in S1 with brewer's yeast and ferment it under microaerobic conditions at a temperature of 26-32 ℃ and an oxygen content of ≤5% for 48-72 h to obtain the product. The fermentation medium in S1 has a pH of 6-7 and contains the following components in parts by weight: 0.8-1.2 parts peppermint, 1-3 parts rice, 0.8-1.2 parts soybean flour, 0.8-1.2 parts tanshinone, 0.2-0.4 parts NaCl, 0.1-0.2 parts K2HPO4, and 60-100 parts water. In S1, the total inoculum amount of Brussels yeast and millet yeast in the fermentation medium is 3wt%-5wt%. In S2, the inoculum amount of brewer's yeast in the product obtained in S1 is 4wt%-10wt%.

2. The preparation method according to claim 1, characterized in that, The microaerobic fermentation described in S1 is carried out at 180–250 rpm.

3. The preparation method according to claim 1, characterized in that, The microaerobic fermentation described in S2 is carried out at 100–300 rpm.

4. The preparation method according to claim 1, characterized in that, After microaerobic fermentation as described in S2, ultrasonication and solid-liquid separation are performed in sequence.

5. The preparation method according to claim 1, characterized in that, Before inoculation, as described in S1, the fermentation medium will be sterilized.

6. The yeast fermentation product prepared by any one of claims 1 to 5.

7. The use of the yeast fermentation product according to claim 6 in the preparation of cosmetics.

8. A cosmetic product, characterized in that, It includes the yeast fermentation product as described in claim 6, and cosmetic excipients.

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