Bacillus velezensis SFKC01 for oil control and acne removal, fermentation product thereof, application and product

Abstract

This invention relates to the field of microbial technology, specifically to a Bacillus vesiculus SFKC01 for oil control and acne treatment, its fermentation products, applications, and products. The Bacillus vesiculus (… Bacillus from Velez The preservation number of SFKC01 is CGMCC No. 34956. The preparation method of Bacillus vesiliflorus SFKC01 fermentation product includes mixing Bacillus vesiliflorus SFKC01 bacterial suspension with traditional Chinese medicine extracts to obtain a mixed solution, adding feed culture medium, mixing thoroughly, and then carrying out secondary fermentation to obtain a traditional Chinese medicine fermentation broth. The broth is then filtered to remove the bacterial residue. This fermentation product has multiple effects, including oil control, acne treatment, and anti-inflammatory soothing. It can significantly inhibit the growth of harmful bacteria related to acne, while promoting the reproduction of beneficial bacteria in the skin, regulating the skin's microecological balance, enhancing the skin barrier function and self-immunity, and improving the problem of recurring acne from the source.

Description

Technical Field

[0001] This invention relates to the field of microbial technology, specifically to a Bacillus vesiculosus SFKC01 for oil control and acne removal, its ferments, applications, and products. Background Technology

[0002] Acne (commonly known as pimples) is a chronic inflammatory skin disease occurring in the pilosebaceous unit, commonly found on the face, chest, and back. Its clinical manifestations include comedones, papules, pustules, and nodules. Due to increased academic and work pressure, acne problems are becoming increasingly prominent, especially among teenagers and young adults, with a trend towards younger onset. Acne formation is closely related to excessive sebum secretion, abnormal follicular keratinization, proliferation of Propionibacterium acnes, inflammatory responses, as well as lifestyle factors, stress, and genetics. Furthermore, oily skin and acne development are mutually reinforcing. Excess sebum not only clogs pores but also promotes an anaerobic local microenvironment, thus facilitating the rapid proliferation of microorganisms such as Propionibacterium acnes. Oily skin buildup can also induce or exacerbate local inflammation, making the pilosebaceous unit more susceptible to infection and redness.

[0003] Currently, acne treatments on the market mainly include acid peels, antibiotic treatments, laser treatments, and products containing colloidal sulfur. Acid products improve symptoms by accelerating keratinocyte turnover, but they can easily cause dry skin, flaking, and barrier damage. Antibiotics are fast-acting, but they are prone to drug resistance, and long-term use can lead to skin-dependent dermatitis. Laser treatment can quickly destroy Propionibacterium acnes and effectively inhibit bacterial growth, with a strong effect, but it is difficult to control acne at its root. Colloidal sulfur has a bactericidal effect, but it has a pungent odor and may cause pigmentation. Overall, existing acne treatments have limited efficacy and significant side effects.

[0004] Currently, there are reports on the application of probiotic fermentation products in the field of skin care. For example, Chinese patent CN115044505B discloses an antimicrobial lipopeptide produced by a strain of Bacillus belyssum and its application in cosmetics and food. It also discloses the extraction of antimicrobial substances from this strain to obtain the Bacillus belyssum antimicrobial lipopeptide, which exhibits significant antimicrobial effects against skin pathogens such as Propionibacterium acnes, Candida albicans, and Staphylococcus aureus. Chinese patent CN120290421B discloses that the fermentation product filtrate of Bacillus belyssum BTN-HB-F4 has good antioxidant, anti-aging, whitening, and skin-soothing effects. It can also inhibit the production of inflammatory factors such as IL-6 and TNF-α, and inhibit the growth of Staphylococcus aureus, demonstrating its ability to regulate the skin microecology.

[0005] While existing research has revealed the potential applications of Bacillus vesiculosus in antibacterial, antioxidant, anti-aging, whitening, and skin-soothing effects, there is a lack of studies specifically addressing the simultaneous needs of oil control, antibacterial, and anti-inflammatory effects for acne-prone skin. Especially in the acne formation mechanism, oil control and acne treatment must work simultaneously to effectively break the vicious cycle between excessive sebum secretion and inflammatory response. Therefore, there is an urgent need to develop a Bacillus vesiculosus fermentation product that can both inhibit acne-related pathogens and improve sebum secretion and inflammatory response to meet the pressing demand of acne sufferers for safe and effective acne treatment products. Summary of the Invention

[0006] In view of the technical problems of existing acne treatments having obvious side effects or limited effects, the present invention provides a Bacillus vesiculosus SFKC01 for oil control and acne removal, its fermentation products, applications and products.

[0007] The specific technical solution is as follows: In a first aspect, the present invention provides a Bacillus belye SFKC01 strain, which was deposited on June 20, 2025, at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 34956, located at No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing.

[0008] Secondly, the present invention provides a Bacillus vesiculus SFKC01 ferment for oil control and acne removal. The preparation method includes mixing the above-mentioned Bacillus vesiculus SFKC01 bacterial liquid with a traditional Chinese medicine extract to obtain a mixed liquid, adding a supplementary culture medium and mixing well, and then carrying out a secondary fermentation to obtain a traditional Chinese medicine fermentation liquid. After filtration to remove the bacterial mud, the Bacillus vesiculus SFKC01 ferment is obtained. The raw materials of the traditional Chinese medicine extract include the following components in parts by weight: Centella asiatica 5-30 parts, chamomile 5-15 parts, kudzu root 5-15 parts, salvia miltiorrhiza 5-15 parts, sophora flavescens 2-10 parts, green tea 2-10 parts, aloe vera 2-10 parts, white willow bark 2-10 parts, coptis chinensis 2-10 parts, and purslane 2-10 parts.

[0009] Based on the synergistic effects and compatibility of different herbal extracts, the herbal ingredients are selected, mainly consisting of ten herbs: Centella asiatica, chamomile, kudzu root, salvia miltiorrhiza, sophora flavescens, aloe vera, green tea, white willow bark, coptis chinensis, and purslane, achieving a scientific compounding. After fermentation by Bacillus vesiculosus SFKC01, the herbal extracts contain multiple active ingredients that promote the proliferation of Bifidobacterium and Staphylococcus epidermidis, inhibit the reproduction of Staphylococcus aureus and Propionibacterium acnes, regulate excessive sebum secretion, balance oil and water, inhibit lipid peroxidation, and reduce sebum blockage leading to acne and closed comedones, thus controlling oil and clearing acne. It effectively inhibits melanin synthesis, reducing melanin deposition at acne sites during the acne treatment process, thereby preventing acne scars and resulting in whiter and more beautiful skin after repair. It is also relatively gentle and non-irritating, making it very suitable for sensitive skin.

[0010] Furthermore, the preparation method of Bacillus belyss SFKC01 bacterial suspension is as follows: Bacillus belyss SFKC01 is inoculated into yeast extract-glucose medium at an inoculation amount of 2%, and cultured at 37°C and 150 rpm for 24 h to obtain Bacillus belyss SFKC01 bacterial suspension.

[0011] Furthermore, the yeast extract-glucose medium is prepared by mixing 20g of glucose and 10g of yeast extract, and adding distilled water to a final volume of 1000mL.

[0012] Furthermore, the weight ratio of Bacillus vesiculosus SFKC01 bacterial solution to traditional Chinese medicine extract is 1:2 to 2:1.

[0013] Furthermore, the specific preparation method of the herbal extract is as follows: weigh the raw materials according to the required weight proportions, mix them, decoct them three times with water, each time adding water 20 times the mass of the herbal medicine, and each decoction time is 40 minutes. Combine the decoctions obtained from the three decoctions, concentrate and filter them to finally obtain a herbal extract that is 10 times the dry weight of the herbal medicine.

[0014] Furthermore, the fed-batch culture medium comprises the following components by weight: 1-3 parts glucose, 1-3 parts yeast extract, 5-15 parts type III collagen peptides, and 10-30 parts acetylated sodium hyaluronate; the amount of fed-batch culture medium added is 60-80 g / 1000 mL of mixture.

[0015] Preferably, the supplemental culture medium is prepared by mixing 10g of glucose, 5g of yeast extract, 10g of type III collagen peptides and 30g of acetylated sodium hyaluronate, and the amount of supplemental culture medium added is 70g / 1000mL of mixture.

[0016] Thirdly, the present invention provides an application of the above-mentioned Bacillus vesicularis SFKC01 or Bacillus vesicularis SFKC01 fermentation product in the preparation of oil-controlling and acne-removing products.

[0017] Fourthly, the present invention also provides an oil-controlling and acne-removing product comprising the above-mentioned Bacillus vesicularis SFKC01 or Bacillus vesicularis SFKC01 ferment.

[0018] Furthermore, the above-mentioned oil-controlling and acne-removing products include the following components in parts by weight: 25-40 parts of Bacillus vesiculosus SFKC01 ferment, 10-30 parts of emulsifier, 10-30 parts of moisturizer, 10-25 parts of emollient, 0-1 part of thickener and 0-1 part of chelating agent.

[0019] Furthermore, the moisturizer is one or more of the following: acetylated sodium hyaluronate, glycerin, allantoin, butylene glycol, arginine, ceramide, panthenol, tremella polysaccharide, glycosyl trehalose, tocopheryl acetate, lysine, serine, glycine, and algae extract, preferably a composition of acetylated sodium hyaluronate, ceramide, and tremella polysaccharide in a mass ratio of 4:3:2.

[0020] Furthermore, the softener is one or more of the following: dioctyl carbonate, shea butter, polydimethyl sulfoxane, cyclodimethylsiloxane, stearyl alcohol, panthenol, methyl glucosyl ether, polyethylene glycol 33, and lanolin alcohol, preferably a composition of stearyl alcohol and lanolin alcohol in a mass ratio of 1:1.

[0021] Furthermore, the thickener is one or more of xanthan gum, carbomer, dextrin, hydroxyethyl cellulose, disodium cocoamphodiacetate, and sodium acrylate, preferably a combination of xanthan gum and carbomer in a mass ratio of 1:1.

[0022] Furthermore, the chelating agent is one or more of disodium EDTA, trisodium EDTA, tetrasodium EDTA, and pentasodium pentetrate, preferably trisodium EDTA.

[0023] Furthermore, the preparation method of the oil-controlling and acne-removing product includes: adding moisturizer and thickener to Bacillus vesiculosus SFKC01 fermentation product, letting it stand for 12 hours, heating to 80°C to obtain an aqueous phase; mixing emulsifier and emollient, heating to dissolve and maintaining at 80°C to obtain an oil phase; adding the oil phase and chelating agent together to the aqueous phase, stirring thoroughly through a homogeneous mixer, and cooling to 30°C to obtain the oil-controlling and acne-removing product.

[0024] The beneficial effects of this invention are as follows: 1. This invention provides a Bacillus vesiculus SFKC01 and its fermentation product, used to prepare a product of traditional Chinese medicine extracts fermented with Bacillus vesiculus SFKC01. This product has multiple effects, including oil control, acne removal, and anti-inflammatory soothing. It can significantly inhibit the growth of harmful bacteria related to acne, while promoting the reproduction of beneficial bacteria in the skin, regulating the skin's microecological balance, enhancing the skin barrier function and self-immunity, and improving the problem of recurring acne from the source. After application, the product has a stable oil control effect, obvious acne removal and fading of acne scars effects, and is gentle and non-irritating, making it suitable for long-term use in cosmetics, significantly improving the overall skin care effect and safety of oil control and acne removal products.

[0025] 2. This invention provides a Bacillus vesiculosus SFKC01 fermentation product, whose main active ingredients include glycolipids (such as rhamnolipids and sophorolipids), bacteriocins, peptides, and amino acids. Among these, glycolipids possess excellent surface activity and biocompatibility, exhibit low irritation, and can improve skin sensitivity, rosacea, and inflammation, while promoting wound repair and the penetration of active ingredients. Bacteriocins, as natural antibacterial active substances, can effectively inhibit the proliferation of Staphylococcus aureus and Propionibacterium acnes, maintaining the stability of the skin's microecology. Peptides and amino acids provide nutritional support to the skin and enhance its repair capabilities. The introduction of acetylated sodium hyaluronate and type III collagen peptides during the secondary fermentation process further enhances the moisturizing and nourishing effects, promotes the expression of proteins related to epidermal differentiation and barrier function, and achieves a synergistic enhancement of acne treatment, repair, and moisturizing.

[0026] 3. This invention is based on the principles of traditional Chinese medicine, the science of dermatology, and the microbial fermentation technology. It achieves the organic integration of active ingredients of Chinese herbal medicine, functional peptides and moisturizing factors through microbial fermentation technology. While ensuring safety, it significantly improves the oil control and acne removal effect, and has good application prospects and promotion value. Attached Figure Description

[0027] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, for those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0028] Figure 1 This is a graph showing the results of an in vitro tyrosinase inhibition rate experiment in Example 11 of this application.

[0029] Figure 2 This is a graph showing the experimental results of the oil inhibition rate in Example 19 of this application. Detailed Implementation

[0030] To enable those skilled in the art to better understand the technical solutions of this invention, the technical solutions of the embodiments of this invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of this invention, and not all embodiments. Based on the embodiments of this invention, all other embodiments obtained by those skilled in the art without creative effort should fall within the scope of protection of this invention.

[0031] Example 1: Isolation and purification of bacterial strains (1) Use a sterile sampling spoon to sample the soil of Yunmenshan. Avoid contamination during the sampling process. Store the samples in a refrigerated box immediately after sampling and record them by number.

[0032] (2) Weigh 1g of soil sample, dissolve it in 10ml of physiological saline, mix well, and then perform a tenfold gradient dilution of the sample liquid. Take 100μL of each gradient and spread it evenly on a plate culture medium. Incubate overnight at 37℃.

[0033] (3) After the culture was completed, colonies of different shapes and sizes grew on the plates. Colonies with the characteristics of being milky white, round and slightly convex, with a smooth surface and viscous texture were selected for microscopic examination. After microscopic examination, spores with straight rod-like shapes were selected for further streak purification. The plates were activated for 1 day, and then liquid culture was carried out at 37℃ and 150 rpm. After the culture was completed, the fermentation broth was centrifuged, and the supernatant was used for surface tension testing. The strain with the highest surface tension was selected for strain identification.

[0034] Example 2: Strain Identification (1) The strain with the highest surface tension was sent to Sangon Biotech (Shanghai) Co., Ltd. for strain identification.

[0035] (2) The primer sequences used in the strain identification process are as follows: 27F: 5'- AGAGTTTGATCMTGGCTCAG -3' (SEQ No: 1); 1492R: 5'-GGTTACCTTGTTACGACTT-3' (SEQ No: 2).

[0036] (3) The 16S rDNA sequence of the strain isolated in Example 1 is as follows:

[0037] (4) The strain was identified as belonging to the genus Bacillus. Bacillus It is presumed to be Bacillus belye Bacillus velezensis .

[0038] (5) The above strain was named Bacillus belyssus SFKC01 and deposited at the China General Microbiological Culture Collection Center (CGMCC). The specific deposit information is as follows: deposit date: June 20, 2025; classification and name: Bacillus belyssus Bacillus velezensis Collection number: CGMCC No.34956, Collection address: No.3, No.1 Courtyard, Beichen West Road, Chaoyang District, Beijing.

[0039] Examples 3-8 Preparation of Bacillus belye SFKC01 fermentation product Examples 3-8 of this invention describe the preparation of Bacillus belyssus SFKC01 fermentation products, specifically including the following steps: (1) Preparation of Chinese herbal extract: Weigh 5-30g of Centella asiatica, 5-15g of Chamomile, 5-15g of Pueraria lobata, 5-15g of Salvia miltiorrhiza, 2-10g of Sophora flavescens, 2-10g of green tea, 2-10g of Aloe vera, 2-10g of white willow bark, 2-10g of Coptis chinensis and 2-10g of Portulaca oleracea, mix them together, decoct them three times with water, each time adding water 20 times the total mass of Chinese herbal medicine, each decoction time is 40min, combine the decoctions obtained from the three decoctions, concentrate and filter them, and finally obtain a Chinese herbal extract that is 10 times the dry weight of Chinese herbal medicine; (2) Preparation of Bacillus belyss SFKC01 bacterial culture: Bacillus belyss SFKC01 was inoculated into the culture medium at an inoculation rate of 2% and cultured at 37℃ and 150rpm for 24h to obtain Bacillus belyss SFKC01 bacterial culture; (3) Preparation of fermentation liquid of Chinese herbal medicine: Bacillus SFKC01 bacterial solution and Chinese herbal medicine extract were mixed at a weight ratio of 1:1. The following components were added to every 1000 mL of the mixture: 10 g glucose, 5 g yeast extract powder, 10 g type III collagen peptide and 30 g acetylated sodium hyaluronate. After mixing, a second fermentation was carried out and cultured at 37℃ and 150 rpm for 18 h to obtain fermentation liquid of Chinese herbal medicine rich in bacterial mud.

[0040] (4) Filtration of fermentation liquid: The fermentation liquid of Chinese herbal medicine rich in bacterial mud is filtered through a cross-flow membrane filter to remove bacterial mud and obtain clear and transparent Bacillus SFKC01 fermentation product.

[0041] The raw materials used in the preparation of the herbal extracts in Examples 3-8 of this invention are shown in Table 1: Table 1. Weights of raw materials used in the preparation of herbal extracts in Examples 3-8

[0042] Comparative Example 1: Fermentation supernatant of Bacillus belye SFKC01 Bacillus berleis SFKC01 was inoculated into the culture medium at a rate of 2% and cultured at 37°C and 150 rpm for 24 h to obtain Bacillus berleis SFKC01 bacterial suspension. After centrifugation to remove the bacterial cells, the fermentation supernatant of Bacillus berleis SFKC01 was obtained.

[0043] Comparative Example 2: Traditional Chinese Medicine Extracts Weigh out 18g of Centella asiatica, 8g of chamomile, 12g of kudzu root, 6g of salvia miltiorrhiza, 5g of sophora flavescens, 10g of green tea, 5g of aloe vera, 5g of white willow bark, 6g of coptis chinensis, and 10g of purslane. Mix them together and decoct them three times with water. Each time, add water that is 20 times the weight of the Chinese herbs. Each decoction time is 40 minutes. Combine the three decoctions, concentrate them, and filter them to obtain a Chinese herbal extract that is 10 times the dry weight of the Chinese herbs.

[0044] Example 9: Antibacterial Experiment of Skin Pathogenic Bacteria (1) Preparation of sample solutions: Take the Bacillus SFKC01 fermentation product obtained in Examples 3-8, the fermentation supernatant of Comparative Example 1 and the herbal extract of Comparative Example 2, and prepare a 10 mg / mL sample solution with sterile distilled water.

[0045] (2) After activating Propionibacterium acnes, Staphylococcus aureus, and Candida albicans, the bacterial solution was diluted to 1.0 × 10⁻⁶. 6 CFU / ml. 100 μL of diluted bacterial solution was added to each well of a 96-well plate. For Examples 3-8 and Comparative Examples 1-2, 100 μL of the corresponding sample solution was added. The positive control group received 100 μL of 0.2% erythromycin solution, and the blank control group received 100 μL of sterile distilled water. Each group was set up in triplicate. After incubating the 96-well plate in an anaerobic incubator for 48 hours, the OD of each well was measured using a microplate reader. 600 The values ​​were used to determine the inhibition rates of each group of sample solutions against Propionibacterium acnes, Staphylococcus aureus, and Candida albicans. The specific results are shown in Table 2.

[0046] Table 2. Antibacterial effects of each group of sample solutions against different harmful bacteria.

[0047] The Bacillus SFKC01 fermentation products prepared in Examples 3-8 all exhibited significant inhibitory activity against Propionibacterium acnes, Staphylococcus aureus, and Candida albicans. This indicates that the fermentation product can not only effectively inhibit the growth of Propionibacterium acnes, reducing the risk of acne from the source, but also inhibit common harmful skin bacteria such as Staphylococcus aureus and Candida albicans, maintaining the ecological balance of the skin surface microbiome. In terms of inhibition rate, Example 4 showed the best antibacterial effect, proving that this traditional Chinese medicine formula is the optimal one.

[0048] Furthermore, the *Bacillus vesiculosus* SFKC01 ferment broth prepared in Example 4 showed significantly better antibacterial effects against *Propionibacterium acnes*, *Staphylococcus aureus*, and *Candida albicans* than Comparative Examples 1 and 2. Compared with simple herbal extracts or *Bacillus vesiculosus* SFKC01 fermentation supernatant, the ferment broth prepared in this invention exhibited stronger broad-spectrum antibacterial activity. This result indicates a significant synergistic effect between herbal extracts and *Bacillus vesiculosus* SFKC01. Through the fermentation process, the effective components in the herbal extracts are further transformed, activated, or enriched. Simultaneously, *Bacillus vesiculosus* SFKC01 can produce antimicrobial peptides, surfactants, and other metabolites with antibacterial functions. The combination of these two significantly increases the number of antibacterial active components in the fermentation system, simultaneously leveraging the anti-inflammatory, heat-clearing, and oil-controlling effects of the herbal components, as well as the strong antibacterial and microecological regulatory effects of the *Bacillus vesiculosus* metabolites, thus forming a synergistic enhancement effect. In summary, this synergistic effect not only gives the fermented product a higher antibacterial strength, but also allows it to exert a lasting conditioning effect at the level of skin microecology, effectively inhibiting the growth of acne-related pathogens and achieving multiple benefits such as oil control, acne removal, and improvement of the skin barrier.

[0049] Example 10: Experiment on the proliferation rate of beneficial bacteria (1) Preparation of sample solutions: Take the Bacillus SFKC01 fermentation product obtained in Examples 3-8, the fermentation supernatant of Comparative Example 1 and the herbal extract of Comparative Example 2, and prepare a 10 mg / mL sample solution with sterile distilled water.

[0050] (2) After activating Bifidobacterium, Staphylococcus epidermidis, and Lactobacillus rhamnosus, the bacterial solutions were diluted to 1.0 × 10⁶ CFU / ml. 100 μL of the diluted bacterial solution was added to each well of a 96-well plate. 100 μL of the corresponding sample solution was added to groups 3-8 and comparative groups 1-2, respectively. 100 μL of sterile distilled water was added to the blank control group. Three replicates were set up for each group. The 96-well plates were incubated in an anaerobic chamber for 48 h. The OD600 value of each well was measured using a microplate reader to determine the proliferation rate of Bifidobacterium, Staphylococcus epidermidis, and Lactobacillus rhamnosus in each group of sample solutions. The specific results are shown in Table 3.

[0051] Table 3. Effects of each group of sample solutions on the proliferation of beneficial bacteria.

[0052] According to the data in Table 3, the Bacillus vesiculus SFKC01 fermentation product prepared in Examples 3-8 has a technical effect on promoting the growth and reproduction of Bifidobacterium, Staphylococcus epidermidis, and Lactobacillus rhamnosus. The Bacillus vesiculus fermentation supernatant prepared in Comparative Example 1 has no significant promoting effect on Bifidobacterium, Staphylococcus epidermidis, and Lactobacillus rhamnosus. The herbal extract prepared in Comparative Example 2 has a significant promoting effect on Bifidobacterium, Staphylococcus epidermidis, and Lactobacillus rhamnosus. The above results indicate that the combination of Bacillus vesiculus fermentation supernatant and herbal extract in this invention can significantly improve the growth of beneficial bacteria (Bifidobacterium, Staphylococcus epidermidis, and Lactobacillus rhamnosus) on the skin surface, greatly enhance the activity of the skin surface microecology, and strengthen the barrier function of the skin surface.

[0053] Example 11 In vitro tyrosinase inhibition rate experiment Set up sample tubes (T2), sample background (T1), enzyme reaction tubes (C2), and solvent background (C1) using 10 mL test tubes. For each sample and each tested concentration, set up three parallel tubes for each sample (T2) and enzyme reaction tube (C2). Add the sample solutions from Examples 3-8, the fermentation supernatant of Comparative Example 1, and the herbal extract from Comparative Example 2 as reference Table 4, mix thoroughly, and incubate in a 37°C water bath for 10 minutes. Add 2 mL of levodopa solution to each tube sequentially, controlling the reaction time for each tube to 5 minutes. Immediately transfer the reaction solutions from each tube to a cuvette and measure the absorbance at 475 nm. Calculate the tyrosinase inhibition rate using the following formula:

[0054] Table 4 Sampling Requirements

[0055] Test results as follows Figure 1 As shown, the inhibition rates of tyrosinase on the fermentation supernatant of Examples 3-8, Comparative Example 1, and the herbal extract of Comparative Example 2 were 49%, 98%, 64.3%, 84.8%, 56.8%, 82.31%, 21.62%, and 79.62%, respectively. This indicates that the herbal fermentation broth provided by the present invention has a high tyrosinase inhibition rate and a certain whitening and spot-fading effect. Among them, Example 4 showed the highest tyrosinase inhibition rate, and Example 4 was selected as the optimal formulation.

[0056] Examples 12-18: Preparation of oil-controlling and acne-removing products Oil-controlling and acne-removing products were prepared using the Bacillus vesiculosus SFKC01 fermentation product from Example 4 of this invention, resulting in Examples 12-18. The specific operation steps are as follows: Take 25-40 parts of the *Bacillus vesiculus* SFKC01 ferment broth prepared in Example 4, 10-30 parts of emulsifier, 10-30 parts of moisturizer, 10-25 parts of emollient, 0-1 part of thickener, and 0-1 part of chelating agent. Add the moisturizer and thickener to the *Bacillus vesiculus* SFKC01 ferment broth, let it stand for 12 hours, and heat to 80°C to obtain the aqueous phase. Mix the emulsifier and emollient, heat to dissolve, and maintain at 80°C to obtain the oil phase. Then add the oil phase and chelating agent together to the aqueous phase, stir thoroughly using a homogeneous mixer, and cool to 30°C to obtain the oil-controlling and acne-removing product.

[0057] The emulsifier is a composition of sorbitan laurate, glyceryl-4 oleate and carnosine in a mass ratio of 4:3:2; the moisturizer is a composition of acetylated sodium hyaluronate, ceramide and tremella polysaccharide in a mass ratio of 4:3:2; the emollient is a composition of stearyl alcohol and lanolin alcohol in a mass ratio of 1:1; the thickener is a composition of xanthan gum and carbomer in a mass ratio of 1:1; and the chelating agent is trisodium EDTA.

[0058] The composition of the oil-controlling and acne-removing products in Examples 12-18 is shown in Table 5.

[0059] Table 5. Composition ratio of oil-controlling and acne-removing products

[0060] Comparative Example 3: An oil-controlling and acne-removing product prepared from Bacillus vesiculosus SFKC01 The difference from Example 16 is that the Bacillus vesiculosus SFKC01 fermentation supernatant prepared in Comparative Example 1 was used instead of the Bacillus vesiculosus SFKC01 fermentation product prepared in Example 4, while all other treatments were the same.

[0061] Comparative Example 4: An oil-controlling and acne-removing product prepared from a traditional Chinese medicine extract. The difference from Example 16 is that the traditional Chinese medicine extract prepared in Comparative Example 2 was used instead of the Bacillus SFKC01 fermentation product prepared in Example 4, while all other treatments were the same.

[0062] Example 19 Anti-inflammatory and soothing effect test Macrophages were stimulated with 1 μg / mL LPS working solution. The changes in the levels of pro-inflammatory factors (IL-1β, TNF-α, IL-6) after sample treatment were detected to evaluate the anti-inflammatory and soothing efficacy of the oil-controlling and acne-removing products prepared in Examples 12-18 and Comparative Examples 3-4. The steps are as follows: (1) With 2.2×10 5Macrophages were seeded into 6-well plates at a seeding density of 1 cell / well. 2 mL of MEM cell basal medium was added to each well. The seeded cell culture plates were then placed in a cell culture incubator (5% CO2, 37°C) and cultured for 24 h.

[0063] (2) Discard the old cell culture medium in the wells. Add 1% of the oil-controlling and acne-removing product aqueous solution prepared in Examples 12-18 or Comparative Examples 3 and 4 to each well of the sample group. Add 1.8 mL of physiological saline to each well as the blank control group.

[0064] (3) After administration, place the 6-well plate in a cell culture incubator (5% CO2, 37℃) for 2 hours.

[0065] (4) LPS stimulation: 2 hours after administration, 200 μL of working solution containing 1 μg / mL LPS was added to each well of each group and placed in a cell culture incubator (5% CO2, 37℃) for 22 hours.

[0066] (5) ELISA detection: Collect cell culture supernatant and detect the levels of IL-1β, TNF-α and IL-6 in the collected cell supernatant according to the instructions of the Elabscience series ELISA kit.

[0067] (6) Calculate the inhibition rate of pro-inflammatory factors for each sample group according to the following formula:

[0068] The specific test results of the inhibition rate of different ratio compositions are shown in Table 6.

[0069] Table 6. Inhibitory effects of different formulations on pro-inflammatory factors

[0070] The results show that the oil-controlling and acne-removing products provided by this invention have an inhibitory effect on inflammatory factors IL-1β, TNF-α, and IL-6. The product from Example 16 showed the best inhibitory effect, effectively inhibiting the release and transmission of inflammatory factors, and playing an anti-inflammatory and soothing role, thereby effectively repairing inflamed skin. The oil-controlling and acne-removing product from Example 16 was selected for subsequent safety evaluation.

[0071] Example 20: Experiment on the detection of oil content The oil-controlling efficacy of the oil-controlling and acne-removing products prepared in Examples 12-18 and Comparative Examples 3 and 4 was evaluated by detecting the changes in oil content of sebaceous gland cells after treatment with different samples.

[0072] 1. Materials: Human sebaceous gland cell line, 96-well cell culture plate, complete cell culture medium, PBS buffer, linoleic acid, Nile red staining solution, isotretinoin (positive control), oil control samples to be tested, ELISA reader, cell culture incubator.

[0073] 2. Dosage concentration: The final concentration of the test sample group and the positive control group (isotretinoin) was 0.001%.

[0074] 3. Experimental Grouping Blank control group: normal sebaceous gland cells, without linoleic acid induction or sample treatment; Model group: sebaceous gland cells + linoleic acid induction (modeling, increased sebum production); Positive control group: sebaceous gland cells + linoleic acid + 0.001% isotretinoin; Test sample group: sebaceous gland cells + linoleic acid + 0.001% test sample.

[0075] 4. Experimental Procedure (1) Cell plating: The extracted sebaceous gland cell suspension was seeded into 96-well plates and cultured for 18-24 hours until the cell confluence reached 80%; (2) Add sample solution: Discard the original culture medium and add 50 μL of the corresponding sample solution to each well; (3) Modeling and culture: 50uL of pure cell culture medium was added to the model group; 50uL of cell culture medium containing linoleic acid was added to the blank control, positive control and sample group respectively. The culture plates were placed in the incubator and cultured for (24±1)h.

[0076] (4) Nile red staining and fluorescence detection: After the culture is completed, wash once with PBS; add 100uL of Nile red staining solution to each well and incubate at 37°C in the dark for 20min; measure the fluorescence intensity with an ELISA reader at Ex / Em=485nm / 565nm.

[0077] (5) Calculate the oil inhibition rate using the following formula:

[0078] MEI 样品 MEI B MEI M The values ​​represent the mean fluorescence intensity of the sample group, blank control group, and model group, respectively.

[0079] Test results as follows Figure 2As shown, the oil inhibition rates of Examples 12-18, Comparative Examples 3 and 4 were 21.23%, 26.5%, 18.24%, 22.89%, 29.61%, 23.41%, 19.17%, 18.39%, 21.58%, and 26.89%, respectively. Among them, Example 16 had the highest oil inhibition rate of 29.61%. The oil-controlling and acne-removing product of Example 16 provided by the present invention has excellent oil inhibition effect.

[0080] Example 21 Skin Irritation Test (1) Animals and rearing environment: Dutch rabbits (Certificate No.: QZTF(Qing)2023-0002), ordinary grade, provided by Tongfu Breeding Farm in Qingzhou City, a total of 6 rabbits. The weight is 2.1~2.2kg. The animals are kept in ordinary grade animal rooms in individual cages. The temperature of the animal room is 20~22℃ and the humidity is 45~55%RH. Animal feed is purchased from Fulikai Pet Products Co., Ltd., and drinking water is tap water.

[0081] (2) Experimental Method: Approximately 24 hours before the experiment, the hair on both sides of the spine on the back of the Dutch rabbits was shaved, with a shaved area of ​​approximately 3cm × 3cm on both the left and right sides. Approximately 1g of the oil-controlling and acne-removing product prepared in Example 16 was applied to the skin on the left side, with an application area of ​​approximately 2.5cm × 2.5cm. It was applied once a day for 14 consecutive days, serving as the experimental group; the skin on the right side served as the control, without any application of any substance. Starting from the second day, the hair was shaved before each application, and the residue was washed off with warm water. The results were observed 1 hour after application, and the skin irritation test was scored according to the "Cosmetic Hygiene Standard". The experimental results are shown in Table 7. The results show that the oil-controlling and acne-removing product prepared in Example 16 was non-irritating to the skin of the Dutch rabbits.

[0082] Table 7 Results of the skin irritation test of oil-controlling and acne-removing products on Dutch rabbits.

[0083] Note: Skin irritation intensity grading: 0~<0.5 No irritation; 0.5~<2.0 Mild irritation; 2.0~<6.0 Moderate irritation; 6.0~<8.0 Strong irritation.

[0084] Example 22 Safety Experiment on Human Skin Safety Experiment: Skin irritation was tested on the oil-controlling and acne-removing product prepared in Example 16. Twenty subjects (10 males and 10 females, aged 20-35 years, with no history of skin diseases) were selected. A patch test strip coated with 0.03 mL of the oil-controlling and acne-removing product was applied to normal skin on the left and right forearms of the subjects for 48 hours. Within 30 minutes of removing the patch, examinations were performed according to the CTFA Safety Assessment Guidelines, for a total of 6 times. On the 6th day after the application period, additional examinations were conducted to observe delayed reactions; the criteria are shown in Table 8.

[0085] Table 8 Comparison of Skin Patch Experiment Results

[0086] The number of people with non-negative test results during the trial is shown in Table 9. This indicates that the oil-controlling and acne-removing treatment in Example 16 has high safety, does not cause allergic reactions, and can be used with confidence.

[0087] Table 9 Results of skin patch experiment

[0088] Although the present invention has been described in detail with reference to the accompanying drawings and preferred embodiments, the present invention is not limited thereto. Various equivalent modifications or substitutions can be made to the embodiments of the present invention by those skilled in the art without departing from the spirit and essence of the invention, and such modifications or substitutions should all be within the scope of the present invention. Any variations or substitutions that can be easily conceived by those skilled in the art within the technical scope disclosed in the present invention should also be covered within the protection scope of the present invention.

Claims

1. A strain of Bacillus belye SFKC01, characterized in that, Bacillus velezensis (Bacillus velezensis) Bacillus velezensis SFKC01 has been deposited with China General Microbiological Culture Collection Center on June 20, 2025, and the deposit number is CGMCC No. 34956, and the deposit address is No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing.

2. A Bacillus vesiculosus SFKC01 fermentation product for oil control and acne removal, characterized in that, The preparation method includes mixing the Bacillus vesicle SFKC01 bacterial solution as described in claim 1 with the traditional Chinese medicine extract to obtain a mixed solution, adding feed culture medium and mixing well, and then carrying out secondary fermentation to obtain the traditional Chinese medicine fermentation broth. After filtration to remove the bacterial mud, the Bacillus vesicle SFKC01 fermentation product is obtained. The raw materials of the traditional Chinese medicine extract include the following components in parts by weight: Centella asiatica 5-30 parts, chamomile 5-15 parts, kudzu root 5-15 parts, salvia miltiorrhiza 5-15 parts, sophora flavescens 2-10 parts, green tea 2-10 parts, aloe vera 2-10 parts, white willow bark 2-10 parts, coptis chinensis 2-10 parts, and purslane 2-10 parts.

3. The Bacillus vesiculus SFKC01 ferment as described in claim 2, characterized in that, The preparation method of Bacillus vesiculosus SFKC01 bacterial suspension is as follows: Bacillus vesiculosus SFKC01 is inoculated into yeast extract-glucose medium at an inoculation amount of 2%, and cultured at 37℃ and 150rpm for 24h to obtain Bacillus vesiculosus SFKC01 bacterial suspension.

4. The Bacillus vesiculus SFKC01 ferment as described in claim 2, characterized in that, The specific preparation method of the herbal extract is as follows: weigh the raw materials according to the required weight proportions, mix them, decoct them three times with water, each time adding water 20 times the mass of the herbal medicine, and each decoction time is 40 minutes. Combine the decoctions obtained from the three decoctions, concentrate and filter them to finally obtain a herbal extract that is 10 times the dry weight of the herbal medicine.

5. The Bacillus vesiculus SFKC01 ferment as described in claim 2, characterized in that, The weight ratio of Bacillus vesiculosus SFKC01 bacterial suspension to traditional Chinese medicine extract is 1:2 to 2:

1.

6. The Bacillus vesiculus SFKC01 ferment as described in claim 2, characterized in that, The fed-batch culture medium consists of the following components by weight: 1-3 parts glucose, 1-3 parts yeast extract, 5-15 parts type III collagen peptides, and 10-30 parts acetylated sodium hyaluronate; the amount of fed-batch culture medium added is 60-80 g / 1000 mL of mixture.

7. The use of Bacillus vesiculosus SFKC01 as described in claim 1 or Bacillus vesiculosus SFKC01 fermentation product as described in any one of claims 2-6 in the preparation of oil-controlling and acne-removing products.

8. An oil-controlling and acne-removing product comprising Bacillus vesiculosus SFKC01 as described in claim 1 or Bacillus vesiculosus SFKC01 fermentation product as described in any one of claims 2-6.

9. The oil-controlling and acne-removing product as described in claim 8, characterized in that, It includes the following components by weight: 25-40 parts of Bacillus vesiculus SFKC01 ferment, 10-30 parts of emulsifier, 10-30 parts of humectant, 10-25 parts of emollient, 0-1 part of thickener and 0-1 part of chelating agent.

10. The oil-controlling and acne-removing product as described in claim 9, characterized in that, The emulsifier is a composition of sorbitan laurate, glyceryl-4 oleate and carnosine in a mass ratio of 4:3:2; the moisturizer is a composition of acetylated sodium hyaluronate, ceramide and tremella polysaccharide in a mass ratio of 4:3:2; the emollient is a composition of stearyl alcohol and lanolin alcohol in a mass ratio of 1:1; the thickener is a composition of xanthan gum and carbomer in a mass ratio of 1:1; and the chelating agent is trisodium EDTA.

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