Compound bacterium fermentation product as well as preparation method and application thereof

Abstract

The invention discloses a compound bacterium fermentation product as well as a preparation method and application thereof. Inoculating the compound bacteria into a fermentation culture medium containing traditional Chinese medicines for fermentation culture to obtain traditional Chinese medicine compound bacteria fermentation liquor; the addition amount of the traditional Chinese medicines is 5-10% of the weight of the fermentation culture medium. The compound bacteria fermentation product prepared by the invention has the effects of resisting oxidation and whitening, the ulva lactuca and the asparagus are fermented by compound bacteria and have an antibacterial effect, the centipeda minima and the coastal angelica sinensis stems are fermented by compound bacteria and have a whitening effect, and combined fermentation and synergistic interaction are realized. In the prepared cosmetic components, the rosa canina fruit oil and the white pool flower oil are combined for use, so that the moisture content, the skin elasticity and the roughness of the skin are improved, and the combination has a synergistic effect.

Description

A fermentation product of a compound bacteria, its preparation method and application Technical Field

[0001] This invention belongs to the field of cosmetic preparation technology, specifically relating to a compound bacterial fermentation product, its preparation method, and its application. Background Technology

[0002] With the increasing demand from consumers for natural, safe, and effective cosmetics, probiotic fermentation technology for traditional Chinese medicine is becoming a research hotspot and important direction in the field of cosmetic raw material development due to its unique advantages. This technology, through the biotransformation effect of probiotics, can significantly enhance the efficacy of active ingredients in traditional Chinese medicine and endow them with new skincare functions, providing an innovative solution for creating "green, natural, and effective" cosmetics.

[0003] The fermentation process of traditional Chinese medicine by probiotics (such as lactic acid bacteria, bifidobacteria, Bacillus licheniformis, etc.) is not only a simple extraction of components, but also a deep biotransformation process. Its core advantages are mainly reflected in three aspects: (1) Enhancing the dissolution and transformation of active ingredients: The enzyme system of probiotics can decompose the cell walls and complex macromolecules of traditional Chinese medicine, making the active ingredients (such as polysaccharides, flavonoids, polyphenols, etc.) easier to dissolve and significantly increasing their content. For example, after fermentation by compound probiotics, the total flavonoid content in Panax notoginseng residue increased by 123.83%. At the same time, the metabolism of bacteria can transform some components into small molecules or new active substances that are more easily absorbed by the skin, such as producing 2,3-butanediol with moisturizing effect. (2) Enhancing and synergizing the original efficacy: Fermentation not only preserves and enhances the whitening, anti-aging and other effects of traditional Chinese medicine itself, but also the probiotics and their metabolites can produce a synergistic effect. Studies have shown that co-culture extracts of Lactobacillus salivarius with Cordyceps militaris or Dendrobium officinale can significantly regulate inflammatory factors and promote the expression of skin collagen, exhibiting excellent anti-inflammatory and repairing effects. Similarly, after fermentation by Lactococcus lactis, the fermentation broth of medicinal Ferula salivarius showed better performance than ordinary extracts in inhibiting sebum secretion and shrinking pores. (3) Reducing potential irritation and realizing resource recycling: The fermentation process can degrade certain macromolecular proteins or impurities in traditional Chinese medicine that may cause skin irritation, making the product milder and safer. In addition, this technology provides a high-value-added resource utilization pathway for by-products (such as dregs) in the production process of traditional Chinese medicine, which is in line with the concept of sustainable development.

[0004] Currently, research in this field has evolved from fermentation of single strains and single medicinal materials to the exploration of synergistic fermentation of compound traditional Chinese medicines using multiple strains, aiming to address more complex sub-health skin issues. Probiotic-fermented traditional Chinese medicines have enormous potential as cosmetic raw materials, but continued research is needed in areas such as the development of synergistic drug combinations, optimization of fermentation processes, and long-term validation of clinical efficacy. Summary of the Invention

[0005] The purpose of this invention is to provide a compound bacterial fermentation product, its preparation method, and its application.

[0006] A compound bacterial fermentation product is obtained by inoculating the compound bacteria into a fermentation medium containing traditional Chinese medicine and fermenting it to obtain a traditional Chinese medicine compound bacterial fermentation broth; the amount of traditional Chinese medicine added is 5-10% of the weight of the fermentation medium.

[0007] The Chinese medicinal ingredients are one or more of the following: Ulva prostrata, Asparagus cochinchinensis, Coriander, Angelica sinensis stem, and Melia toosendan bark.

[0008] The compound bacteria are one or more of the following: Lactobacillus acidophilus, Lactobacillus bulgaricus, Lactobacillus plantarum, Lactobacillus casei, Streptococcus thermophilus, yeast, and Bacillus subtilis.

[0009] The fermentation medium comprises: 8-12 g / L peptone, 6-10 g / L beef extract, 3-7 g / L yeast powder, 15-25 g / L glucose, 1-3 g / L dipotassium hydrogen phosphate, 1-3 g / L diammonium hydrogen citrate, 3-7 g / L sodium acetate, 0.1-0.3 g / L magnesium sulfate, 0.02-0.06 g / L manganese sulfate, and 0.5-1.5 g / L Tween 80, with the pH adjusted to 5-7.

[0010] The application of the fermentation products of the compound bacteria in the preparation of skin-whitening and brightening cosmetics.

[0011] The method for preparing skin-whitening and brightening cosmetics using the fermentation products of the aforementioned compound bacteria is carried out according to the following steps:

[0012] (1) Take 50-100 parts water, 3-8 parts propylene glycol and 1-3 parts trehalose according to the weight, mix them, stir and adjust the pH to 6.5-7.2; (2) Add 3-8 parts of compound bacterial fermentation product, 0.5-3 parts vitamin E and 5-15 parts emulsifier, and stir evenly; (3) Add 3-8 parts of vegetable oil and 3-5 parts of vegetable gum, homogenize and make.

[0013] The emulsifier is one or more of cetearyl alcohol, cetearyl glucoside, glyceryl stearate, sorbitan monooleate, and polydimethylsiloxane copolyol.

[0014] The vegetable oil is one or more of the following: meadowfoam oil, sunflower seed oil, peony seed oil, rice bran oil, shea butter, Brazilian peach fruit oil, sweet orange fruit oil, Wenzhou mandarin fruit oil, Rosa rugosa fruit oil, and coconut fruit oil.

[0015] The plant gum is one or more of gum arabic, guar gum, frankincense gum, benzoin gum, undecenoyl xanthan gum, and cinnamon gum.

[0016] The stirring rate in step (1) is 50-100 rpm and the time is 5-10 min; the stirring rate in step (2) is 200-500 rpm and the time is 5-10 min; the homogenization rate in step (3) is 3000-5000 rpm and the time is 3-5 min.

[0017] The beneficial effects of this invention are as follows: The compound bacterial fermentation product prepared by this invention has antibacterial and whitening effects. Ulva and Asparagus, after compound bacterial fermentation, exhibit antibacterial effects, while Coriander and Angelica sinensis stems, after compound bacterial fermentation, exhibit whitening effects. Furthermore, the combined fermentation process has a synergistic effect. In the cosmetic components prepared by this invention, the combined use of Rosa canina fruit oil and meadowfoam oil improves skin hydration, elasticity, and roughness, and the combination also has a synergistic effect. Detailed Implementation

[0018] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0019] Example 1: A compound bacterial fermentation product. The compound bacteria were inoculated into a fermentation medium containing traditional Chinese medicine (TCM) and fermented to obtain the TCM compound bacterial fermentation broth. The amount of TCM added was 8% of the weight of the fermentation medium. The TCM components were a mixture of Ulva lactuca and Asparagus cochinchinensis in a 1:1 mass ratio. The compound bacteria were a mixture of Lactobacillus bulgaricus, yeast, and Bacillus subtilis in a 2:1:1 viable count ratio. The total inoculation amount of the compound bacteria was 2% of the weight of the fermentation medium, and the total viable count of the compound bacterial inoculation broth was 2 x 10⁻⁶. 8 CFU / mL. The fermentation temperature was 37℃, and the fermentation time was 3 days. After fermentation, the fermentation broth was centrifuged at 8000 rpm for 15 min, the supernatant was collected, and the filtrate was obtained after sterilization by filtering through a 0.22-micron filter membrane. The fermentation filtrate was concentrated to 1 / 6 of its original volume to prepare the compound bacterial fermentation product.

[0020] The fermentation medium comprises: 10 g / L peptone, 8 g / L beef extract, 5 g / L yeast powder, 20 g / L glucose, 2 g / L dipotassium hydrogen phosphate, 2 g / L diammonium hydrogen citrate, 5 g / L sodium acetate, 0.2 g / L magnesium sulfate, 0.04 g / L manganese sulfate, and 1 g / L Tween 80, with the pH adjusted to 6.2.

[0021] The preparation method of the skin whitening and brightening cosmetic using the fermentation product of the compound bacteria is carried out according to the following steps: (1) Take 75 parts water, 6 parts propylene glycol and 2 parts trehalose according to the weight ratio, mix them, stir at 80 rpm for 8 min, and adjust the pH to 6.8; (2) Add 6 parts of the fermentation product of the compound bacteria, 2 parts vitamin E, 5 parts cetearyl alcohol and 5 parts cetearyl glucoside, stir at 300 rpm for 8 min; (3) Add 3 parts sunflower seed oil, 3 parts peony seed oil, 2 parts gum arabic and 2 parts guar gum, homogenize at 4000 rpm for 4 min, and make the product.

[0022] Example 2: A compound bacterial fermentation product. The compound bacteria were inoculated into a fermentation medium containing traditional Chinese medicine (TCM) and fermented to obtain the TCM compound bacterial fermentation broth. The amount of TCM added was 6% of the weight of the fermentation medium. The TCM was a mixture of *Coriandrum sativum* and *Angelica sinensis* stems in a mass ratio of 2:1. The compound bacteria were a mixture of *Lactobacillus plantarum*, yeast, and *Bacillus subtilis* in a viable count ratio of 1:1:1. The total inoculation amount of the compound bacteria was 2.5% of the weight of the fermentation medium, and the total viable count of the compound bacterial inoculation broth was 1.5 x 10⁻⁶. 8 CFU / mL. The fermentation temperature was 37℃, and the fermentation time was 3 days. After fermentation, the fermentation broth was centrifuged at 8000 rpm for 15 min, the supernatant was collected, and the filtrate was obtained after sterilization by passing it through a 0.22-micron filter membrane. The fermentation filtrate was concentrated to 1 / 5 of its original volume to prepare the compound bacterial fermentation product.

[0023] The fermentation medium comprises: 10 g / L peptone, 8 g / L beef extract, 5 g / L yeast powder, 20 g / L glucose, 2 g / L dipotassium hydrogen phosphate, 2 g / L diammonium hydrogen citrate, 5 g / L sodium acetate, 0.2 g / L magnesium sulfate, 0.04 g / L manganese sulfate, and 1 g / L Tween 80, with the pH adjusted to 6.5.

[0024] The preparation method of the skin-whitening and brightening cosmetic using the fermentation product of the compound bacteria is carried out according to the following steps: (1) Take 60 parts of water, 4 parts of propylene glycol and 1 part of trehalose according to the weight ratio, mix them, stir at 60 rpm for 6 min, and adjust the pH to 6.6; (2) Add 5 parts of the fermentation product of the compound bacteria, 1.5 parts of vitamin E and 7.5 parts of glyceryl stearate, and stir at 300 rpm for 6 min; (3) Add 4 parts of rice bran oil and 3 parts of benzoin gum, and homogenize at 3500 rpm for 3 min to make the product.

[0025] Example 3 describes a compound bacterial fermentation product. The compound bacteria were inoculated into a fermentation medium containing *Melia azedarach* bark and fermented to obtain the fermentation broth containing the traditional Chinese medicine compound bacteria. The amount of *Melia azedarach* bark added was 8% of the weight of the fermentation medium. The compound bacteria were a mixture of *Lactobacillus casei*, yeast, and *Bacillus subtilis* in a 1:1:1 ratio of viable cells. The total inoculation amount of the compound bacteria was 1.5% of the weight of the fermentation medium, and the total viable cell count of the compound bacteria inoculation broth was 1.5 x 10⁻⁶. 8 CFU / mL. The fermentation temperature was 37℃, and the fermentation time was 4 days. After fermentation, the fermentation broth was centrifuged at 8000 rpm for 15 min, the supernatant was collected, and the filtrate was obtained after sterilization by passing it through a 0.22-micron filter membrane. The fermentation filtrate was concentrated to 1 / 7 of its original volume to prepare the compound bacterial fermentation product.

[0026] The fermentation medium comprises: 10 g / L peptone, 8 g / L beef extract, 5 g / L yeast powder, 20 g / L glucose, 2 g / L dipotassium hydrogen phosphate, 2 g / L diammonium hydrogen citrate, 5 g / L sodium acetate, 0.2 g / L magnesium sulfate, 0.04 g / L manganese sulfate, and 1 g / L Tween 80, with the pH adjusted to 6.9.

[0027] The preparation method of the skin-whitening and brightening cosmetic using the fermentation product of the compound bacteria is carried out according to the following steps: (1) Take 90 parts of water, 6 parts of propylene glycol and 3 parts of trehalose according to the weight ratio, mix them, stir at 90 rpm for 9 min, and adjust the pH to 7.0; (2) Add 8 parts of the fermentation product of the compound bacteria, 2.5 parts of vitamin E and 12 parts of dehydrated sorbitan monooleate, and stir at 400 rpm for 8 min; (3) Add 4 parts of meadowfoam oil, 4 parts of rosehip oil, and 5 parts of purple wormwood, and homogenize at 5000 rpm for 5 min to prepare the product.

[0028] Comparative Example 1: A compound bacterial fermentation product was prepared by inoculating the compound bacteria into a fermentation medium containing Ulva lactuca and fermenting to obtain the fermentation broth of the traditional Chinese medicine compound bacteria. The amount of Ulva lactuca added was 8% of the weight of the fermentation medium. The compound bacteria were a mixture of Lactobacillus bulgaricus, yeast, and Bacillus subtilis in a viable count ratio of 2:1:1. The total inoculation amount of the compound bacteria was 2% of the weight of the fermentation medium, and the total viable count of the compound bacteria inoculation broth was 2 x 10⁻⁶. 8 CFU / mL. The fermentation temperature was 37℃, and the fermentation time was 3 days. After fermentation, the fermentation broth was centrifuged at 8000 rpm for 15 min, the supernatant was collected, and the filtrate was obtained after sterilization by filtering through a 0.22-micron filter membrane. The fermentation filtrate was concentrated to 1 / 6 of its original volume to prepare the compound bacterial fermentation product.

[0029] The fermentation medium comprises: 10 g / L peptone, 8 g / L beef extract, 5 g / L yeast powder, 20 g / L glucose, 2 g / L dipotassium hydrogen phosphate, 2 g / L diammonium hydrogen citrate, 5 g / L sodium acetate, 0.2 g / L magnesium sulfate, 0.04 g / L manganese sulfate, and 1 g / L Tween 80, with the pH adjusted to 6.2.

[0030] The preparation method of the skin whitening and brightening cosmetic using the fermentation product of the compound bacteria is carried out according to the following steps: (1) Take 75 parts water, 6 parts propylene glycol and 2 parts trehalose according to the weight ratio, mix them, stir at 80 rpm for 8 min, and adjust the pH to 6.8; (2) Add 6 parts of the fermentation product of the compound bacteria, 2 parts vitamin E, 5 parts cetearyl alcohol and 5 parts cetearyl glucoside, stir at 300 rpm for 8 min; (3) Add 3 parts sunflower seed oil, 3 parts peony seed oil, 2 parts gum arabic and 2 parts guar gum, homogenize at 4000 rpm for 4 min, and make the product.

[0031] Comparative Example 2 describes a fermentation product of a compound microbial culture. The compound microbial culture was inoculated into a fermentation medium containing asparagus and fermented to obtain the fermentation broth of the traditional Chinese medicine compound microbial culture. The amount of asparagus added was 8% of the weight of the fermentation medium. The compound microbial culture was a mixture of Lactobacillus bulgaricus, yeast, and Bacillus subtilis in a viable count ratio of 2:1:1. The total inoculation amount of the compound microbial culture was 2% of the weight of the fermentation medium, and the total viable count of the compound microbial inoculation broth was 2 x 10⁻⁶. 8 CFU / mL. The fermentation temperature was 37℃, and the fermentation time was 3 days. After fermentation, the fermentation broth was centrifuged at 8000 rpm for 15 min, the supernatant was collected, and the filtrate was obtained after sterilization by filtering through a 0.22-micron filter membrane. The fermentation filtrate was concentrated to 1 / 6 of its original volume to prepare the compound bacterial fermentation product.

[0032] The fermentation medium comprises: 10 g / L peptone, 8 g / L beef extract, 5 g / L yeast powder, 20 g / L glucose, 2 g / L dipotassium hydrogen phosphate, 2 g / L diammonium hydrogen citrate, 5 g / L sodium acetate, 0.2 g / L magnesium sulfate, 0.04 g / L manganese sulfate, and 1 g / L Tween 80, with the pH adjusted to 6.2.

[0033] The preparation method of the skin whitening and brightening cosmetic using the fermentation product of the compound bacteria is carried out according to the following steps: (1) Take 75 parts water, 6 parts propylene glycol and 2 parts trehalose according to the weight ratio, mix them, stir at 80 rpm for 8 min, and adjust the pH to 6.8; (2) Add 6 parts of the fermentation product of the compound bacteria, 2 parts vitamin E, 5 parts cetearyl alcohol and 5 parts cetearyl glucoside, stir at 300 rpm for 8 min; (3) Add 3 parts sunflower seed oil, 3 parts peony seed oil, 2 parts gum arabic and 2 parts guar gum, homogenize at 4000 rpm for 4 min, and make the product.

[0034] Comparative Example 3 describes a fermentation product made from a compound microbial culture medium. The compound microbial culture was inoculated into a fermentation medium containing *Coriandrum sativum* and fermented to obtain the fermentation broth. The amount of *Coriandrum sativum* added was 6% of the weight of the fermentation medium. The compound microbial culture was a mixture of *Lactobacillus plantarum*, yeast, and *Bacillus subtilis* in a 1:1:1 ratio of viable cells. The inoculation amount of the compound microbial culture was 2.5% of the weight of the fermentation medium, and the total viable cell count of the inoculated broth was 1.5 x 10⁻⁶. 8 CFU / mL. The fermentation temperature was 37℃, and the fermentation time was 3 days. After fermentation, the fermentation broth was centrifuged at 8000 rpm for 15 min, the supernatant was collected, and the filtrate was obtained after sterilization by passing it through a 0.22-micron filter membrane. The fermentation filtrate was concentrated to 1 / 5 of its original volume to prepare the compound bacterial fermentation product.

[0035] The fermentation medium comprises: 10 g / L peptone, 8 g / L beef extract, 5 g / L yeast powder, 20 g / L glucose, 2 g / L dipotassium hydrogen phosphate, 2 g / L diammonium hydrogen citrate, 5 g / L sodium acetate, 0.2 g / L magnesium sulfate, 0.04 g / L manganese sulfate, and 1 g / L Tween 80, with the pH adjusted to 6.5.

[0036] The preparation method of the skin-whitening and brightening cosmetic using the fermentation product of the compound bacteria is carried out according to the following steps: (1) Take 60 parts of water, 4 parts of propylene glycol and 1 part of trehalose according to the weight ratio, mix them, stir at 60 rpm for 6 min, and adjust the pH to 6.6; (2) Add 5 parts of the fermentation product of the compound bacteria, 1.5 parts of vitamin E and 7.5 parts of glyceryl stearate, and stir at 300 rpm for 6 min; (3) Add 4 parts of rice bran oil and 3 parts of benzoin gum, and homogenize at 3500 rpm for 3 min to make the product.

[0037] Comparative Example 4 describes a fermentation product made from a compound microbial culture medium. The compound microbial culture was inoculated into a fermentation medium containing *Angelica sinensis* stems and fermented to obtain the fermentation broth. The amount of *Angelica sinensis* stems added was 6% of the weight of the fermentation medium. The compound microbial culture was a mixture of *Lactobacillus plantarum*, yeast, and *Bacillus subtilis* in a 1:1:1 ratio of viable cells. The total inoculation amount of the compound microbial culture was 2.5% of the weight of the fermentation medium, and the total viable cell count of the inoculated broth was 1.5 x 10⁻⁶. 8 CFU / mL. The fermentation temperature was 37℃, and the fermentation time was 3 days. After fermentation, the fermentation broth was centrifuged at 8000 rpm for 15 min, the supernatant was collected, and the filtrate was obtained after sterilization by passing it through a 0.22-micron filter membrane. The fermentation filtrate was concentrated to 1 / 5 of its original volume to prepare the compound bacterial fermentation product.

[0038] The fermentation medium comprises: 10 g / L peptone, 8 g / L beef extract, 5 g / L yeast powder, 20 g / L glucose, 2 g / L dipotassium hydrogen phosphate, 2 g / L diammonium hydrogen citrate, 5 g / L sodium acetate, 0.2 g / L magnesium sulfate, 0.04 g / L manganese sulfate, and 1 g / L Tween 80, with the pH adjusted to 6.5.

[0039] The preparation method of the skin-whitening and brightening cosmetic using the fermentation product of the compound bacteria is carried out according to the following steps: (1) Take 60 parts of water, 4 parts of propylene glycol and 1 part of trehalose according to the weight ratio, mix them, stir at 60 rpm for 6 min, and adjust the pH to 6.6; (2) Add 5 parts of the fermentation product of the compound bacteria, 1.5 parts of vitamin E and 7.5 parts of glyceryl stearate, and stir at 300 rpm for 6 min; (3) Add 4 parts of rice bran oil and 3 parts of benzoin gum, and homogenize at 3500 rpm for 3 min to make the product.

[0040] Comparative Example 5 describes a fermentation product of a compound microbial culture. The compound microbial culture was inoculated into a fermentation medium containing *Melia azedarach* bark and fermented to obtain the fermentation broth of the traditional Chinese medicine compound microbial culture. The amount of *Melia azedarach* bark added was 8% of the weight of the fermentation medium. The compound microbial culture was a mixture of *Lactobacillus casei*, yeast, and *Bacillus subtilis* in a 1:1:1 ratio of viable cells. The total inoculation amount of the compound microbial culture was 1.5% of the weight of the fermentation medium, and the total viable cell count of the inoculated broth was 1.5 x 10⁻⁶. 8 CFU / mL. The fermentation temperature was 37℃, and the fermentation time was 4 days. After fermentation, the fermentation broth was centrifuged at 8000 rpm for 15 min, the supernatant was collected, and the filtrate was obtained after sterilization by passing it through a 0.22-micron filter membrane. The fermentation filtrate was concentrated to 1 / 7 of its original volume to prepare the compound bacterial fermentation product.

[0041] The fermentation medium comprises: 10 g / L peptone, 8 g / L beef extract, 5 g / L yeast powder, 20 g / L glucose, 2 g / L dipotassium hydrogen phosphate, 2 g / L diammonium hydrogen citrate, 5 g / L sodium acetate, 0.2 g / L magnesium sulfate, 0.04 g / L manganese sulfate, and 1 g / L Tween 80, with the pH adjusted to 6.9.

[0042] The preparation method of the skin-whitening and brightening cosmetic using the fermentation product of the compound bacteria is carried out according to the following steps: (1) Take 90 parts of water, 6 parts of propylene glycol and 3 parts of trehalose according to the weight ratio, mix them, stir at 90 rpm for 9 min, and adjust the pH to 7.0; (2) Add 8 parts of the fermentation product of the compound bacteria, 2.5 parts of vitamin E and 12 parts of dehydrated sorbitan monooleate, and stir at 400 rpm for 8 min; (3) Add 8 parts of meadowfoam oil and 5 parts of purple shellac, and homogenize at 5000 rpm for 5 min to make the product.

[0043] Comparative Example 6 describes a fermentation product made from a compound microbial culture medium. The compound microbial culture was inoculated into a fermentation medium containing *Melia azedarach* bark and fermented to obtain the fermentation broth containing the traditional Chinese medicine compound microbial culture. The amount of *Melia azedarach* bark added was 8% of the weight of the fermentation medium. The compound microbial culture was a mixture of *Lactobacillus casei*, yeast, and *Bacillus subtilis* in a 1:1:1 ratio of viable cells. The inoculation amount of the compound microbial culture was 1.5% of the weight of the fermentation medium, and the total viable cell count of the inoculated broth was 1.5 x 10⁻⁶. 8 CFU / mL. The fermentation temperature was 37℃, and the fermentation time was 4 days. After fermentation, the fermentation broth was centrifuged at 8000 rpm for 15 min, the supernatant was collected, and the filtrate was obtained after sterilization by passing it through a 0.22-micron filter membrane. The fermentation filtrate was concentrated to 1 / 7 of its original volume to prepare the compound bacterial fermentation product.

[0044] The fermentation medium comprises: 10 g / L peptone, 8 g / L beef extract, 5 g / L yeast powder, 20 g / L glucose, 2 g / L dipotassium hydrogen phosphate, 2 g / L diammonium hydrogen citrate, 5 g / L sodium acetate, 0.2 g / L magnesium sulfate, 0.04 g / L manganese sulfate, and 1 g / L Tween 80, with the pH adjusted to 6.9.

[0045] The preparation method of the skin-whitening and brightening cosmetic using the fermentation product of the compound bacteria is carried out according to the following steps: (1) Take 90 parts of water, 6 parts of propylene glycol and 3 parts of trehalose according to the weight ratio, mix them, stir at 90 rpm for 9 min, and adjust the pH to 7.0; (2) Add 8 parts of the fermentation product of the compound bacteria, 2.5 parts of vitamin E and 12 parts of dehydrated sorbitan monooleate, and stir at 400 rpm for 8 min; (3) Add 8 parts of Rosa canis fruit oil and 5 parts of purple shellac, and homogenize at 5000 rpm for 5 min to prepare the product.

[0046] Experiment Example 1: Antibacterial Experiment. Activated Candida albicans, Staphylococcus aureus, and Malassezia were prepared into bacterial concentrations of 10 using culture media. 6 CFU / mL bacterial suspension; the fermentation products of the compound bacteria prepared in Example 1 and Comparative Examples 1-2 were prepared with sterile water to form a sample solution with a concentration of 15 wt%; 0.5 ml of bacterial suspension was placed in a test tube, 4.5 ml of sample solution was injected, the mixture was quickly mixed and the time was started immediately. At the 15 min time point, 1.0 ml of the mixture was taken, and the viable bacteria count was determined by viable bacteria culture counting method. Each sample was inoculated into 3 petri dishes, and the inhibition rate was calculated. The results are shown in Table 1: Table 1

[0047] Note: The comparison with Example 1 group showed P<0.05.

[0048] Example 2: Tyrosinase Inhibition Rate Detection. Phosphate Buffer: Take 250 mL of 0.2 mol / L potassium dihydrogen phosphate solution, add 118 mL of 0.2 mol / L sodium hydroxide solution, and dilute to 1000 mL with deionized water. L-Tyrosine Solution: Weigh 27.17 mg of L-tyrosine, dissolve in phosphate buffer, and dilute to 100 mL in a volumetric flask to prepare a 1.5 mmol / L L-tyrosine solution. Tyrosinase Solution: Weigh 18.5 mg of tyrosinase (25 KU), add 12.5 mL of phosphate buffer to dilute to 2 KU / mL, aliquot, and freeze. Dilute to 200 U / mL with phosphate buffer before use.

[0049] The fermentation products of the compound bacteria prepared in Example 2, Comparative Examples 3 and 4 were taken, and phosphate buffer, sample, tyrosine, and tyrosinase were added respectively under light-protected conditions. The mixtures were incubated at 37°C for 10 min in a microplate reader, and the absorbance was measured at 475 nm. Each measurement required four groups: sample, blank, A0, and A1, with two replicates per group, as follows: A0: 50 μL tyrosine solution + 100 μL buffer + 50 μL tyrosinase; A1: 50 μL tyrosine solution + 150 μL buffer; Sample: 50 μL tyrosine solution + 50 μL sample + 50 μL buffer + 50 μL tyrosinase; Blank: 50 μL tyrosine solution + 50 μL sample + 100 μL buffer.

[0050] Tyrosinase inhibition rate = (1 - (sample - blank) / (A0 - A1)) 100%.

[0051] The experimental results were statistically analyzed, and the quantitative data were used... x ± (mean ± standard deviation) represents the mean. The Kolmogorov-Smirnov test was used to test the normality of the data. For normally distributed data, a t-test was used to compare the differences in means between two groups, with P < 0.05 considered statistically significant. The results are shown in Table 2.

[0052] Note: The comparison with Example 2 group showed P<0.05.

[0053] Experimental Example 3: The cosmetics prepared in Example 3, Comparative Example 5, and Comparative Example 6 were used. Forty-five female volunteers aged 22-45 years were selected on a voluntary basis and randomly divided into groups of 15. Each subject applied 10mg of the cosmetic to both sides of their face twice a day, morning and evening, for a total of 2 weeks. Before the test, the subjects washed the test area with clean water, dried it, and exposed the test area. They sat quietly for 20 minutes in an environment of 25±1℃ and 50±5% relative humidity. The blank group applied 10mg of clean water. The stratum corneum moisture content of the test area was measured using a skin moisture test probe, and the average value of 5 measurements was taken. The stratum corneum moisture content was measured before the start of the experiment. On the evening of the 14th day, 1 hour after the sample was used, the stratum corneum moisture content of the subjects was measured, and the skin moisture content change rate (%) was calculated.

[0054] Similarly, skin elasticity and roughness were measured using a skin elasticity tester before the start of the experiment. On day 14, 1 hour after the sample was used, the skin elasticity and roughness of the test subjects were measured, and the rate of change (%) of skin elasticity and roughness was calculated.

[0055] Skin moisture content change rate (%) = (moisture content after use - moisture content before use) / moisture content before use × 100%; Skin elasticity change rate (%) = (skin elasticity after use - skin elasticity before use) / skin elasticity before use × 100%; Roughness change rate (%) = (roughness before use - roughness after use) / roughness before use × 100%.

[0056] The experimental results were statistically analyzed, and the quantitative data were used... x ± (mean ± standard deviation) represents the mean. The Kolmogorov-Smirnov test was used to test the normality of the data. For normally distributed data, a t-test was used to compare the differences in means between two groups, with P < 0.05 considered statistically significant. The results are shown in Table 3.

[0057] Note: The comparison with Example 3 group showed P<0.05.

[0058] The above embodiments are merely illustrative examples of preferred embodiments of the present invention and do not encompass all possible implementations. Any modifications and refinements made by those skilled in the art without departing from the spirit and scope of the present invention are considered to fall within the protection scope of the claims.

Claims

1. A fermentation product of a compound microorganism, characterized in that, The compound bacteria are inoculated into a fermentation medium containing traditional Chinese medicine and fermented to obtain a fermentation broth of traditional Chinese medicine compound bacteria; the amount of traditional Chinese medicine added is 5-10% of the weight of the fermentation medium.

2. The fermentation product of the compound bacteria according to claim 1, characterized in that, The Chinese medicinal ingredients are one or more of the following: Ulva prostrata, Asparagus cochinchinensis, Coriander, Angelica sinensis stem, and Melia toosendan bark.

3. The fermentation product of the compound bacteria according to claim 1, characterized in that, The compound bacteria are one or more of the following: Lactobacillus acidophilus, Lactobacillus bulgaricus, Lactobacillus plantarum, Lactobacillus casei, Streptococcus thermophilus, yeast, and Bacillus subtilis.

4. The fermentation product of the compound bacteria according to claim 1, characterized in that, The fermentation medium comprises: 8-12 g / L peptone, 6-10 g / L beef extract, 3-7 g / L yeast powder, 15-25 g / L glucose, 1-3 g / L dipotassium hydrogen phosphate, 1-3 g / L diammonium hydrogen citrate, 3-7 g / L sodium acetate, 0.1-0.3 g / L magnesium sulfate, 0.02-0.06 g / L manganese sulfate, and 0.5-1.5 g / L Tween 80, with the pH adjusted to 5-7.

5. The application of the compound bacterial fermentation product according to claim 1 in the preparation of skin-whitening and brightening cosmetics.

6. A method for preparing a skin-whitening and brightening cosmetic using the fermentation product of the compound bacteria described in claim 1, characterized in that, The following steps are to be followed: (1) Take 50-100 parts of water, 3-8 parts of propylene glycol and 1-3 parts of trehalose according to the weight, mix them, stir and adjust the pH to 6.5-7.2; (2) Add 3-8 parts of compound bacterial fermentation product, 0.5-3 parts of vitamin E and 5-15 parts of emulsifier, and stir evenly; (3) Add 3-8 parts of vegetable oil and 3-5 parts of vegetable gum, homogenize and make.

7. The method for preparing the skin-whitening and brightening cosmetic according to claim 6, characterized in that, The emulsifier is one or more of cetearyl alcohol, cetearyl glucoside, glyceryl stearate, sorbitan monooleate, and polydimethylsiloxane copolyol.

8. The method for preparing the skin-whitening and brightening cosmetic according to claim 6, characterized in that, The vegetable oil is one or more of the following: meadowfoam oil, sunflower seed oil, peony seed oil, rice bran oil, shea butter, Brazilian peach fruit oil, sweet orange fruit oil, Wenzhou mandarin fruit oil, Rosa rugosa fruit oil, and coconut fruit oil.

9. The method for preparing the skin-whitening and brightening cosmetic according to claim 6, characterized in that, The plant gum is one or more of gum arabic, guar gum, frankincense gum, benzoin gum, undecenoyl xanthan gum, and cinnamon gum.

10. The method for preparing the skin-whitening and brightening cosmetic according to claim 6, characterized in that, The stirring rate in step (1) is 50-100 rpm and the time is 5-10 min; the stirring rate in step (2) is 200-500 rpm and the time is 5-10 min; the homogenization rate in step (3) is 3000-5000 rpm and the time is 3-5 min.

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