Saccharomycetes fermentation product with permeation enhancing effect as well as preparation method and application of saccharomycetes fermentation product

Abstract

The invention provides a saccharomycetes fermentation product with a permeation enhancing effect as well as a preparation method and application of the saccharomycetes fermentation product. The preparation method comprises the following steps: firstly, inoculating Brussell bouquet yeast and schizosaccharomyces pombe in a mass ratio of (0.7-1.3): (0.7-1.3) into a fermentation culture medium with specific components, performing micro-aerobic fermentation for 48-72 hours at the temperature of 25-35 DEG C and the oxygen content of less than or equal to 10%, then inoculating saccharomyces cerevisiae, and performing micro-aerobic fermentation for 48-72 hours at the temperature of 26-32 DEG C and the oxygen content of less than or equal to 5%. According to the invention, the bruxel bouquet yeast, the schizosaccharomyces pombe and the saccharomyces cerevisiae are subjected to mixed fermentation in a specific culture medium according to a specific sequence, and compared with products obtained by other fermentation modes, the obtained yeast fermentation product has a more excellent permeation promoting effect; the transdermal absorption of active ingredients in the cosmetic or medical composition can be obviously promoted, and the method is suitable for preparing the cosmetic or medical composition.

Description

Technical Field

[0001] The present invention belongs to the technical field of microbial fermentation, and more specifically, relates to a yeast fermentation product with penetration-promoting effect, a preparation method and application thereof. Background Art

[0002] The skin is a barrier between the human body and the external environment, playing an important role in helping to block the invasion of harmful substances from the outside world and preventing the loss of water or nutrients. However, it is this strong barrier function that becomes an obstacle for the skin to absorb the active ingredients of cosmetics. Therefore, in order to overcome the skin barrier function and promote the active ingredients to enter the deep layers of the skin, thereby playing various physiological regulatory roles on skin cells, it is often necessary to add certain penetration enhancers when preparing cosmetics to enhance the transdermal absorption effect of the active ingredients.

[0003] However, existing chemical penetration enhancers, such as azone, surfactants, organic acids, etc., are all irritating, easily cause skin allergies, and are not suitable for long-term use. Therefore, researchers have gradually shifted their research focus to microbial fermentation products with safe and friendly ingredients. Microorganisms may produce secondary metabolites such as biological enzymes during the fermentation process, which can decompose biological macromolecules such as proteins and polysaccharides into substances with smaller molecular weights, thereby playing a role in promoting penetration. However, the fermentation products obtained by different microorganisms and different fermentation methods are significantly different, and the penetration-promoting effects vary greatly. Summary of the invention

[0004] The present invention aims to provide a method for preparing a yeast fermentation product by mixing and fermenting Brettanomyces brusselsii, Schizosaccharomyces pombe and Saccharomyces cerevisiae in a specific order in a specific culture medium. The yeast fermentation product obtained has a more excellent penetration-enhancing 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.

[0005] The first object of the present invention is to provide a method for preparing a yeast fermentation product.

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

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

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

[0009] The above-mentioned purpose of the present invention is achieved through the following technical solutions:

[0010] The present invention provides a method for preparing a yeast fermentation product, comprising the following steps:

[0011] S1. Inoculate Brettanomyces brusselsii and Schizosaccharomyces pombe in a fermentation medium at a mass ratio of 0.7-1.3:0.7-1.3, and ferment at a temperature of 25-35°C and an oxygen content of ≤10% for 48-72 hours;

[0012] S2. Inoculate the product obtained in S1 with brewer's yeast, and ferment at a temperature of 26 to 32°C and an oxygen content of ≤5% for 48 to 72 hours;

[0013] Among them, the pH of the fermentation medium in S1 is 6-7, and it contains the following components in parts by weight: 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 salvia miltiorrhiza, 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 Brettanomyces brusselsii to Schizosaccharomyces pombe in S1 is 1:1.

[0015] Preferably, the pH of the fermentation medium in S1 is 6-7, and comprises the following components in parts by weight: 1 part of mint, 2 parts of rice, 1 part of soybean powder, 1 part of salvia miltiorrhiza, 0.3 parts of NaCl, 0.15 parts of K2HPO4, and 80 parts of water.

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

[0017] More preferably, the sterilization is performed at 115-125°C for 15-25 min, most preferably at 121°C for 20 min.

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

[0019] The Brussels yeast and Schizosaccharomyces pombe can be activated in a conventional manner before inoculation. For example, the Brussels yeast is cultured in a YPDA medium to a cell density of 1×10 7 ~1×10 8 cfu / mL, Schizosaccharomyces pombe was cultured in CM0181 medium to a cell density of 1×10 7 ~1×10 8 cfu / mL.

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

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

[0022] The brewer's yeast can be activated in a conventional manner before inoculation, such as culturing in PDB medium to a cell density of 1×10 7 ~1×10 8 cfu / mL.

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

[0024] Preferably, after the microaerobic fermentation in S2, ultrasound and solid-liquid separation are performed in sequence.

[0025] Further preferably, the ultrasound is performed at 0.05-0.15 MPa for 1.8-2.2 h.

[0026] More preferably, stirring is performed simultaneously with the ultrasound, such as stirring at 100-200 rpm.

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

[0028] More preferably, the particle size of the filtrate obtained by the filtration is ≤10 μm.

[0029] More preferably, sterilization is performed after the solid-liquid separation.

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

[0031] More preferably, after the sterilization, the temperature is further lowered, such as to below 45°C.

[0032] The yeast fermentation product prepared by the above method has a high content of small molecule polypeptides and amino acids and other components, has a strong transdermal penetration promoting effect, can significantly promote the transdermal absorption of active ingredients in cosmetic or medical compositions, and better play the original efficacy of cosmetic or medical compositions. Therefore, the yeast fermentation product prepared by the above method, the use of the yeast fermentation product in the preparation of cosmetic or medical compositions, and a cosmetic or medical composition comprising the yeast fermentation product and cosmetic or medically acceptable excipients should all be within the protection scope of the present invention.

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

[0034] The present invention performs mixed fermentation of Brettanomyces brusselsii, Schizosaccharomyces pombe and Saccharomyces cerevisiae in a specific order in a specific culture medium. The yeast fermentation product obtained has a more excellent penetration-enhancing 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. DETAILED DESCRIPTION

[0035] The present invention is further described below with reference to specific examples, but the examples do not limit the present invention in any form. Unless otherwise specified, the reagents, methods and equipment used in the present invention are conventional reagents, methods and equipment in the art.

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

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

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

[0039] Activation of Saccharomyces cerevisiae: Culture 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. The fermentation medium was sterilized at 121° C. for 20 min, and then Brettanomyces brusselsii and Schizosaccharomyces pombe were inoculated into the fermentation medium at a mass ratio of 1:1 (so that the total inoculation amount of Brettanomyces brusselsii and Schizosaccharomyces pombe in the fermentation medium was 4 wt %), and the fermentation was carried out in a constant temperature shaker at a temperature of 30° C., an oxygen content of 8%, and a rotation speed of 220 rpm for 60 h;

[0042] S2. Inoculate the product obtained from S1 with Saccharomyces cerevisiae (so that the inoculation amount of Saccharomyces cerevisiae in the product obtained from S1 is 7wt%), and ferment in a constant temperature shaker at 29°C, 3% oxygen content, and 200rpm for 60h, then ultrasonicate at 0.1MPa and 150rpm for 2h, and filter with a 10μm filter membrane. Sterilize the filtrate at 85°C for 25min, and then cool to 45°C.

[0043] Among them, the pH of the fermentation medium in S1 is 6.5, and it contains the following components in parts by mass: 1 part of mint, 2 parts of rice, 1 part of soybean powder, 1 part of salvia miltiorrhiza, 0.3 parts of NaCl, 0.15 parts of K2HPO4, and 80 parts of water.

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

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

[0046] S2. Inoculate the product obtained in S1 with brewer's yeast (so that the inoculation amount of brewer's yeast in the product obtained in S1 is 10wt%), ferment in a constant temperature shaker at 26°C, 5% oxygen content, and 300rpm for 48h, then ultrasonicate at 0.05MPa and 200rpm for 1.8h, filter with a 10μm filter membrane, sterilize the filtrate at 80°C for 30min, and then cool to 45°C;

[0047] Among them, the pH of the fermentation medium in S1 is 7, and it contains the following components in parts by mass: 0.8 parts of mint, 3 parts of rice, 0.8 parts of soybean powder, 0.8 parts of salvia miltiorrhiza, 0.4 parts of NaCl, 0.1 parts of K2HPO4, and 60 parts of water.

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

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

[0050] S2. Inoculate the product obtained from S1 with brewer's yeast (so that the inoculation amount of brewer's yeast in the product obtained from S1 is 4wt%), ferment in a constant temperature shaker at 32°C, 2% oxygen content, and 100rpm for 72h, then ultrasonicate at 0.15MPa and 100rpm for 2.2h, filter with a 10μm filter membrane, sterilize the filtrate at 90°C for 20min, and then cool to 45°C;

[0051] Among them, the pH of the fermentation medium in S1 is 6, and it contains the following components in parts by mass: 1.2 parts of mint, 1 part of rice, 1.2 parts of soybean powder, 1.2 parts of salvia miltiorrhiza, 0.2 parts of NaCl, 0.2 parts of K2HPO4, and 100 parts of water.

[0052] Comparative Example 1

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

[0054] That is, this comparative example is as follows:

[0055] S1. The fermentation medium was sterilized at 121°C for 20 min, and then Saccharomyces cerevisiae was inoculated into the fermentation medium (so that the inoculation amount of Saccharomyces cerevisiae in the fermentation medium was 7 wt%), and the fermentation was carried out in a constant temperature shaker at 29°C, 3% oxygen content, and 200 rpm for 60 h under microaerobic conditions;

[0056] S2. Inoculate the product obtained in S1 with Brettanomyces brusselsii and Schizosaccharomyces pombe in a mass ratio of 1:1 (so that the total inoculation amount of Brettanomyces brusselsii and Schizosaccharomyces pombe in the product obtained in S1 is 4wt%), and ferment in a constant temperature shaker at a temperature of 30°C, an oxygen content of 8%, and a rotation speed of 220rpm for 60h, then ultrasonicate at 0.1MPa and 150rpm for 2h, and filter with a 10μm filter membrane, and sterilize the filtrate at 85°C for 25min, and then cool to 45°C;

[0057] Among them, the pH of the fermentation medium in S1 is 6.5, and it contains the following components in parts by mass: 1 part of mint, 2 parts of rice, 1 part of soybean powder, 1 part of salvia miltiorrhiza, 0.3 parts of NaCl, 0.15 parts of K2HPO4, and 80 parts of water.

[0058] Comparative Example 2

[0059] The same as Example 1, except that the Brussels yeast was replaced by Candida albicans. That is, the specific details of this comparative example are as follows:

[0060] S1. The fermentation medium was sterilized at 121°C for 20 min, and then Candida albicans and Schizosaccharomyces pombe were inoculated into the fermentation medium at a mass ratio of 1:1 (so that the total inoculation amount of Candida albicans and Schizosaccharomyces pombe in the fermentation medium was 4wt%), and the fermentation was carried out in a constant temperature shaker at a temperature of 30°C, an oxygen content of 8%, and a rotation speed of 220 rpm for 60 h;

[0061] S2. Inoculate the product obtained from S1 with Saccharomyces cerevisiae (so that the inoculation amount of Saccharomyces cerevisiae in the product obtained from S1 is 7wt%), and ferment in a constant temperature shaker at 29°C, 3% oxygen content, and 200rpm for 60h, then ultrasonicate at 0.1MPa and 150rpm for 2h, and filter with a 10μm filter membrane. Sterilize the filtrate at 85°C for 25min, and then cool to 45°C.

[0062] Among them, the pH of the fermentation medium in S1 is 6.5, and it contains the following components in parts by mass: 1 part of mint, 2 parts of rice, 1 part of soybean powder, 1 part of salvia miltiorrhiza, 0.3 parts of NaCl, 0.15 parts of K2HPO4, and 80 parts of water.

[0063] Comparative Example 3

[0064] Same as Example 1, except that Schizosaccharomyces pombe is replaced by Torulopsis albus. That is, the specific details of this comparative example are as follows:

[0065] S1. The fermentation medium was sterilized at 121° C. for 20 min, and then Brettanomyces brusselsii and Torulopsis albicans were inoculated into the fermentation medium at a mass ratio of 1:1 (so that the total inoculation amount of Brettanomyces brusselsii and Torulopsis albicans in the fermentation medium was 4 wt %), and the fermentation was carried out in a constant temperature shaker at a temperature of 30° C., an oxygen content of 8%, and a rotation speed of 220 rpm for 60 h;

[0066] S2. Inoculate the product obtained from S1 with Saccharomyces cerevisiae (so that the inoculation amount of Saccharomyces cerevisiae in the product obtained from S1 is 7wt%), and ferment in a constant temperature shaker at 29°C, 3% oxygen content, and 200rpm for 60h, then ultrasonicate at 0.1MPa and 150rpm for 2h, and filter with a 10μm filter membrane. Sterilize the filtrate at 85°C for 25min, and then cool to 45°C.

[0067] Among them, the pH of the fermentation medium in S1 is 6.5, and it contains the following components in parts by mass: 1 part of mint, 2 parts of rice, 1 part of soybean powder, 1 part of salvia miltiorrhiza, 0.3 parts of NaCl, 0.15 parts of K2HPO4, and 80 parts of water.

[0068] Comparative Example 4

[0069] The same as Example 1, except that Saccharomyces cerevisiae is replaced by Hansenula anomala. That is, the specific details of this comparative example are as follows:

[0070] S1. The fermentation medium was sterilized at 121° C. for 20 min, and then Brettanomyces brusselsii and Schizosaccharomyces pombe were inoculated into the fermentation medium at a mass ratio of 1:1 (so that the total inoculation amount of Brettanomyces brusselsii and Schizosaccharomyces pombe in the fermentation medium was 4 wt %), and the fermentation was carried out in a constant temperature shaker at a temperature of 30° C., an oxygen content of 8%, and a rotation speed of 220 rpm for 60 h;

[0071] S2. The abnormal Hansen yeast was inoculated into the product obtained in S1 (the inoculation amount of abnormal Hansen yeast in the product obtained in S1 was 7wt%), and the fermentation was carried out in a constant temperature shaker at a temperature of 29°C, an oxygen content of 3%, and 200rpm for 60h, and then ultrasonicated at 0.1MPa and 150rpm for 2h, and filtered with a 10μm filter membrane. The obtained filtrate was sterilized at 85°C for 25min and then cooled to 45°C;

[0072] Among them, the pH of the fermentation medium in S1 is 6.5, and it contains the following components in parts by mass: 1 part of mint, 2 parts of rice, 1 part of soybean powder, 1 part of salvia miltiorrhiza, 0.3 parts of NaCl, 0.15 parts of K2HPO4, and 80 parts of water.

[0073] Comparative Example 5

[0074] The same as Example 1, except that S1 was inoculated with only Brettanomyces brusselsii. That is, the specific details of this comparative example are as follows:

[0075] S1. The fermentation medium was sterilized at 121° C. for 20 min, and then the Brussels yeast was inoculated into the fermentation medium (so that the inoculation amount of Brussels yeast in the fermentation medium was 4 wt%), and the fermentation was carried out in a constant temperature shaker at a temperature of 30° C., an oxygen content of 8%, and a rotation speed of 220 rpm for 60 h;

[0076] S2. Inoculate the product obtained from S1 with Saccharomyces cerevisiae (so that the inoculation amount of Saccharomyces cerevisiae in the product obtained from S1 is 7wt%), and ferment in a constant temperature shaker at 29°C, 3% oxygen content, and 200rpm for 60h, then ultrasonicate at 0.1MPa and 150rpm for 2h, and filter with a 10μm filter membrane. Sterilize the filtrate at 85°C for 25min, and then cool to 45°C.

[0077] Among them, the pH of the fermentation medium in S1 is 6.5, and it contains the following components in parts by mass: 1 part of mint, 2 parts of rice, 1 part of soybean powder, 1 part of salvia miltiorrhiza, 0.3 parts of NaCl, 0.15 parts of K2HPO4, and 80 parts of water.

[0078] Comparative Example 6

[0079] The same as Example 1, except that S1 was inoculated with only Schizosaccharomyces pombe. That is, the specifics of this comparative example are as follows:

[0080] S1. The fermentation medium was sterilized at 121°C for 20 min, and 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 the fermentation was carried out in a constant temperature shaker at a temperature of 30°C, an oxygen content of 8%, and a rotation speed of 220 rpm for 60 h;

[0081] S2. Inoculate the product obtained from S1 with Saccharomyces cerevisiae (so that the inoculation amount of Saccharomyces cerevisiae in the product obtained from S1 is 7wt%), and ferment in a constant temperature shaker at 29°C, 3% oxygen content, and 200rpm for 60h, then ultrasonicate at 0.1MPa and 150rpm for 2h, and filter with a 10μm filter membrane. Sterilize the filtrate at 85°C for 25min, and then cool to 45°C.

[0082] Among them, the pH of the fermentation medium in S1 is 6.5, and it contains the following components in parts by mass: 1 part of mint, 2 parts of rice, 1 part of soybean powder, 1 part of salvia miltiorrhiza, 0.3 parts of NaCl, 0.15 parts of K2HPO4, and 80 parts of water.

[0083] Comparative Example 7

[0084] The same as Example 1, except that the rice in the fermentation medium is replaced by millet. That is, the specific details of this comparative example are as follows:

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

[0086] S2. Inoculate the product obtained from S1 with Saccharomyces cerevisiae (so that the inoculation amount of Saccharomyces cerevisiae in the product obtained from S1 is 7wt%), and ferment in a constant temperature shaker at 29°C, 3% oxygen content, and 200rpm for 60h, then ultrasonicate at 0.1MPa and 150rpm for 2h, and filter with a 10μm filter membrane. Sterilize the filtrate at 85°C for 25min, and then cool to 45°C.

[0087] Among them, the pH of the fermentation medium in S1 is 6.5, and it contains the following components in parts by mass: 1 part of mint, 2 parts of millet, 1 part of soybean powder, 1 part of salvia miltiorrhiza, 0.3 parts of NaCl, 0.15 parts of K2HPO4, and 80 parts of water.

[0088] Test Example 1: Penetration-enhancing effect of yeast fermentation products

[0089] The back skin of a 2-month-old suckling pig was fixed between the supply tank and the receiving tank of a Franz vertical double-chamber osmotic diffusion cell, 12 mL of normal saline was added to the supply tank, and the receiving tank was not filled with any liquid. After standing for 30 minutes, if there was no significant titration or outflow of subcutaneous fluid on the receiving tank side, it proved that the skin barrier function was intact, otherwise it was judged that the skin barrier function was impaired. After confirming that the skin barrier function was intact, the sample to be tested was added to the receiving tank (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). Then the Franz vertical double chamber osmotic diffusion cell was placed in a 37°C constant temperature water bath, and a sample was taken from the receiving cell at the 24th hour of the water bath, and the content of vitamin A therein was determined by HPLC chromatography (Vertex chromatographic column, eluent was acetonitrile, methanol, and dichloromethane in a volume ratio of 3:3:4, the flow rate of the eluent was 1.0 mL / min, and the column temperature was 25°C). The transdermal absorption rate of the sample to be tested was calculated according to the formula "transdermal absorption rate (%) = vitamin A mass in the receiving cell / vitamin A mass in the sample to be tested × 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 products obtained in Examples 1 to 3 is significantly higher than that in Comparative Examples 1 to 7, indicating that the yeast fermentation products obtained by mixed fermentation of Brettanomyces brussels, Schizosaccharomyces pombe and Saccharomyces cerevisiae in a specific order in a specific culture medium have a more excellent penetration-enhancing effect than the products obtained by other fermentation methods, can significantly promote the transdermal absorption of active ingredients in cosmetic or medical compositions, and are suitable for preparing cosmetic or medical compositions.

[0093] Test Example 2: Transdermal Ability of Essence Containing Yeast Fermentation Products

[0094] 1. Preparation of Essence Containing Yeast Fermentation Products

[0095] (1)Ingredients

[0096] EDTA-2Na 0.05wt%, glycerol 1.00wt%, 1,3-butylene glycol 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%, cyclopolydimethylsiloxane 2.00wt%, hydroxyethyl acrylate / sodium acryloyldimethyl taurate copolymer 0.35wt%, yeast fermentation product obtained in Examples 1 to 3 and Comparative Examples 1 to 7 3.00wt%, and make up to 100wt% with water.

[0097] (2) Preparation method

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

[0099] 2. Preparation of Control Essence

[0100] (1)Ingredients

[0101] EDTA-2Na 0.05wt%, glycerol 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%, cyclopolydimethylsiloxane 2.00wt%, hydroxyethyl acrylate / sodium acryloyldimethyl taurate copolymer 0.35wt%, and make up to 100wt% with water.

[0102] (2) Preparation method

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

[0104] 3. DPPH free radical scavenging experiment

[0105] Sample group: Dissolve the essence in distilled water (so that the final concentration of the essence is 1% (v / v)), take 100 μL and add it to a 96-well plate, then add 100 μL of 0.1 mM DPPH solution, shake for 10 minutes in the dark, and then use an ELISA reader to measure the absorbance Ax at 520 nm. Set up three replicate wells and take the average of the results.

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

[0107] The DPPH free radical scavenging rate of the essence was calculated according to the formula "DPPH free radical scavenging rate (%) = [(A0-Ax) / A0] x 100%". 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 essence containing the yeast fermentation product obtained in Examples 1 to 3 is significantly higher than that of the essence containing the yeast fermentation product obtained in Comparative Examples 1 to 7 and the control essence, indicating that the yeast fermentation product obtained by mixed fermentation of Brussels yeast, Schizosaccharomyces pombe and Saccharomyces cerevisiae in a specific order in a specific culture medium has a more excellent penetration-enhancing effect than the products obtained by other fermentation methods, can significantly promote the transdermal absorption of active ingredients in cosmetics or medical compositions, and is suitable for preparing cosmetics or medical compositions.

[0112] The above embodiments are preferred implementation modes of the present invention, but the implementation modes of the present invention are not limited to the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications that do not deviate from the spirit and principles of the present invention should be equivalent replacement methods and are included in the protection scope of the present invention.

Claims

1. A method for preparing a yeast fermentation product, characterized in that: The steps include: S1. Inoculate Brettanomyces brusselsii and Schizosaccharomyces pombe in a fermentation medium at a mass ratio of 0.7-1.3:0.7-1.3, and ferment at a temperature of 25-35°C and an oxygen content of ≤10% for 48-72 hours; S2. Inoculate the product obtained in S1 with brewer's yeast, and ferment at a temperature of 26 to 32°C and an oxygen content of ≤5% for 48 to 72 hours; Among them, the pH of the fermentation medium in S1 is 6-7, and it contains the following components in parts by weight: 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 salvia miltiorrhiza, 0.2-0.4 parts of NaCl, 0.1-0.2 parts of K2HPO4, and 60-100 parts of water.

2. The preparation method according to claim 1, characterized in that: In S1, the total inoculation amount of Brettanomyces brusselsii and Schizosaccharomyces pombe in the fermentation medium is 3 wt % to 5 wt %.

3. The preparation method according to claim 1, characterized in that: In S2, the inoculation amount of brewer's yeast in the product obtained in S1 is 4wt% to 10wt%.

4. The preparation method according to claim 1, characterized in that: S1 The microaerobic fermentation is carried out at 180-250 rpm.

5. The preparation method according to claim 1, characterized in that: S2 The microaerobic fermentation is carried out at 100-300 rpm.

6. The preparation method according to claim 1, characterized in that: After the microaerobic fermentation in S2, ultrasound and solid-liquid separation are performed in sequence.

7. The preparation method according to claim 1, characterized in that: Before inoculation as described in S1, the fermentation medium is sterilized.

8. A yeast fermentation product prepared by the method according to any one of claims 1 to 7.

9. Use of the yeast fermentation product according to claim 8 in the preparation of cosmetic or medical compositions.

10. A cosmetic or medical composition, characterized in that: The invention comprises the yeast fermentation product according to claim 8, and cosmetically or medically acceptable auxiliary materials.

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