Bio-based cosmetic composition containing fermentation product of bifidobacterium adolescentis and application of bio-based cosmetic composition

Abstract

The invention relates to a bio-based cosmetic composition containing a fermentation product of Bifidobacterium adolescentis and application thereof. The bio-based cosmetic composition disclosed by the invention not only can regulate and maintain skin micro-ecological balance, but also has the effects of relieving skin, resisting wrinkles, tightening skin and the like, compared with a cosmetic with a single effect, the bio-based cosmetic composition can more comprehensively solve skin problems, higher safety and reliability are ensured due to the pure natural characteristic of the bio-based cosmetic composition, and in conclusion, the bio-based cosmetic composition has a very good application prospect. The bio-based cosmetic composition has a good application prospect in the field of cosmetics.

Description

Technical Field

[0001] The invention relates to the field of microorganisms, and in particular to a bio-based cosmetic composition containing a fermentation product of Bifidobacterium adolescentis and application thereof. Background Art

[0002] Traditional cosmetics generally contain a variety of chemical ingredients. Although they may show significant effects in the short term, long-term use may cause irritation or even damage to the skin, and will inevitably impose a certain burden on the environment. As society's awareness of health and environmental protection increases, people are paying more and more attention to the relationship between skin health and skin microecological balance. Many studies have revealed that the imbalance of microbial communities on the surface of the skin may be closely related to a variety of skin diseases and skin aging. In view of this, regulating and maintaining the balance of skin microecology has become a new research hotspot in the field of beauty and skin care.

[0003] Bio-based cosmetics refer to cosmetics made from renewable biological resources (such as plant extracts, microorganisms and their fermentation products). They are gradually gaining favor among consumers, mainly due to their increasing attention to the environmental protection and sustainability of skin care products. The core advantage of bio-based cosmetics is that they rely on natural resources and significantly reduce the dependence on petroleum derivatives. These natural ingredients are usually milder, more suitable for sensitive skin, and have excellent skin affinity.

[0004] Although the application of probiotics in regulating the human intestinal microecology is relatively mature, its application in skin microecology regulation is not yet widespread, especially products with microecological bio-based formulas are even rarer. Summary of the invention

[0005] Purpose of the Invention

[0006] The object of the present invention is to provide a bio-based cosmetic composition and its application in preparing skin care products with skin soothing, anti-wrinkle and / or firming effects.

[0007] Solution

[0008] To achieve the purpose of the present invention, the present invention adopts the following technical scheme.

[0009] In a first aspect, the present invention provides a cosmetic composition, characterized in that the active ingredient of the cosmetic composition comprises:

[0010] (1) The fermentation product of Bifidobacterium adolescentis YG3912; wherein the taxonomic name of Bifidobacterium adolescentis YG3912 is Bifidobacterium adolescentis, which is deposited in the General Microbiological Center of China Microbiological Culture Collection Administration, with a deposit date of June 20, 2024 and a deposit number of CGMCC No.31020;

[0011] (2) Lactobacillus fermentation products;

[0012] (3) Ginseng root extract;

[0013] (4) Purslane extract.

[0014] In a feasible embodiment, the fermentation product of Bifidobacterium adolescentis YG3912 is its fermentation supernatant and optionally a lysate extracted from its fermentation product.

[0015] Preferably, the fermentation supernatant of Bifidobacterium adolescentis YG3912 is prepared by the following method:

[0016] The Bifidobacterium adolescentis YG3912 is fermented and cultured, the fermentation liquid is centrifuged, and the supernatant is collected and filtered to obtain the product.

[0017] Preferably, the lysate of Bifidobacterium adolescentis YG3912 is extracted by the following method:

[0018] The Bifidobacterium adolescentis YG3912 is fermented and cultured, the fermentation broth is centrifuged, the bacteria are collected, the bacteria are resuspended in PBS and the cell walls are broken by ultrasound or enzymatic hydrolysis to obtain a lysate; preferably, the obtained lysate is sterilized.

[0019] In a feasible embodiment, the lactobacillus fermentation product is a fermentation product of Lactobacillus reuteri YG3708 with a preservation number of CGMCC No. 31243; the Lactobacillus reuteri YG3708 is deposited in the General Microbiology Center of the China National Microbiological Culture Collection Administration, with a preservation date of July 9, 2024 and a preservation number of CGMCC No. 31243;

[0020] Preferably, the fermentation product of Lactobacillus reuteri YG3708 is its fermentation supernatant;

[0021] Further preferably, the fermentation supernatant of Lactobacillus reuteri YG3708 is prepared by the following method:

[0022] The Lactobacillus reuteri YG3708 is fermented and cultured, the fermentation liquid is centrifuged, and the supernatant is collected and filtered to obtain the product.

[0023] Preferably, the cosmetic composition further comprises:

[0024] (5) Tremella fuciformis extract;

[0025] (6) Osmanthus extract.

[0026] Preferably, the ginseng root extract, purslane extract, tremella extract and osmanthus extract are all water extracts.

[0027] In a preferred embodiment, the cosmetic composition comprises, by weight percentage:

[0028] 0.1-90% of the fermentation supernatant of Bifidobacterium adolescentis YG3912;

[0029] 0.1-5% lysate of Bifidobacterium adolescentis YG3912;

[0030] 0.1-10% lactobacillus fermentation products;

[0031] 0.05-10% ginseng root extract;

[0032] 0.1-10% purslane extract;

[0033] 0.01-1% Tremella fuciformis extract;

[0034] 0.01-1% osmanthus extract;

[0035] The remainder is cosmetic base.

[0036] Further preferably, the cosmetic composition comprises, by weight percentage:

[0037] 1-50% of the fermentation supernatant of Bifidobacterium adolescentis YG3912;

[0038] 1-5% lysate of Bifidobacterium adolescentis YG3912;

[0039] 1-10% lactobacillus fermentation products;

[0040] 0.05-1% ginseng root extract;

[0041] 0.1-5% purslane extract;

[0042] 0.1-1% Tremella fuciformis extract;

[0043] 0.05-1% osmanthus extract;

[0044] The remainder is cosmetic base.

[0045] More preferably, the cosmetic composition comprises, by weight percentage:

[0046] 1-30% of the fermentation supernatant of Bifidobacterium adolescentis YG3912;

[0047] 1-2% lysate of Bifidobacterium adolescentis YG3912;

[0048] 5-10% lactobacillus fermentation products;

[0049] 0.05-0.1% ginseng root extract;

[0050] 0.1-1% purslane extract;

[0051] 0.1-1% Tremella fuciformis extract;

[0052] 0.05-0.1% osmanthus extract;

[0053] The remainder is cosmetic base.

[0054] Possibly, the cosmetic base is selected from: water, . . .

[0055] In a second aspect, the present invention provides a use of the cosmetic composition as described in the first aspect above in preparing a skin care product having skin soothing, anti-wrinkle and / or firming effects;

[0056] Preferably, the skin care product is a toner, lotion, cream, mask, essence, disposable lotion or eye cream.

[0057] In a third aspect, the present invention provides a method for soothing skin, anti-wrinkle and / or tightening skin, comprising: applying an effective amount of the cosmetic composition as described in the first aspect above to a subject in need thereof.

[0058] Beneficial Effects

[0059] Compared with the prior art, the bio-based cosmetic composition of the present invention has the following obvious advantages:

[0060] (1) Optimizing the balance of skin microecology: The bio-based cosmetic composition of the present invention specifically adjusts the balance of microbial flora that is consistent with the skin characteristics of the Chinese population; specifically, the composition can effectively promote the proliferation of six resident bacteria that are beneficial to skin health, while inhibiting the excessive growth of pathogenic bacteria and Moraxella osloensis, thereby achieving precise regulation of skin microecology; this helps to maintain the stability of the skin's microbial barrier function, thereby significantly enhancing the skin's natural protective ability.

[0061] (2) Pure bio-based formula: This product contains no chemical ingredients at all, embodying the concept of green, environmental protection and health; it avoids irritating chemicals that may be contained in traditional cosmetics, such as fragrances, pigments, preservatives, etc., which may cause allergic reactions; the pure natural characteristics of this product ensure higher safety and reliability, perfectly matching the pursuit of modern consumers for natural and environmentally friendly cosmetics.

[0062] (3) Sustainable development advantage: The bio-based nature of the product ensures the sustainability of the raw material source. Compared with traditional cosmetic raw materials that rely on limited chemical resources or non-renewable resources, the cultivation and production of Bifidobacterium adolescentis achieved through biotechnology, showing excellent environmental friendliness and sustainable utilization of resources, contributing to promoting the cosmetics industry towards a green and sustainable development path.

[0063] (4) Comprehensive skin care effects: The cosmetic composition of the present invention combines multiple functions such as maintaining skin microecological balance, soothing skin, anti-wrinkle and firming. Compared with cosmetics with only a single function, it can solve skin problems more comprehensively. It meets consumers' demand for comprehensive skin care products and provides a more convenient and efficient skin care experience. BRIEF DESCRIPTION OF THE DRAWINGS

[0064] One or more embodiments are exemplarily described by the pictures in the accompanying drawings, and these exemplary descriptions do not constitute limitations on the embodiments. The special word "exemplary" here means "used as an example, embodiment or illustrative". Any embodiment described as "exemplary" here is not necessarily interpreted as being superior or better than other embodiments.

[0065] Figure 1 Shown is the effect of S3912 on the proliferation of S. aureus and C. acnes subsp. acnes as detected by disk diffusion.

[0066] Figure 2 Shown is the effect of S3912 on the proliferation of M. osloensis as measured by disk diffusion.

[0067] Figure 3 Shown are the effects of S3912 on the proliferation of S. hominis, C. simulans, S. capitis, and S. epidermidis as assayed by disk diffusion.

[0068] Figure 4 Shown are the effects of S3912 on the proliferation of C. acnes subsp. defendens and C. granulosum as assayed by disk diffusion.

[0069] Figure 5The figure shows the effect of S3912 on the proliferation of the above 9 strains in BHI liquid culture medium; where "*" represents P < 0.05, and "****" represents P < 0.0001.

[0070] Figure 6 The results of coaggregation experiments between inactivated bacteria of strain YG3912 and S. aureus, C. acnes subsp. acnes and M. osloensis are shown.

[0071] Figure 7 The results of the DPPH free radical scavenging rate (A), elastase inhibition rate (B) and hyaluronidase inhibition rate (C) of the YG3912 fermentation product are shown.

[0072] Figure 8 The DPPH free radical scavenging rate test results of the cosmetic compositions of Preparation Examples 1-4 and Comparative Examples 1-4 are shown; wherein, "*" represents P < 0.05, and "***" represents P < 0.001.

[0073] Fig. 9 The elastase inhibition rate test results of the cosmetic compositions of Preparation Examples 1-4 and Comparative Examples 1-4 are shown; wherein, "*" represents P < 0.05, "**" represents P < 0.01, and "****" represents P < 0.0001.

[0074] Fig.10 The hyaluronidase inhibition rate test results of the cosmetic compositions of Preparation Examples 1-4 and Comparative Examples 1-4 are shown; wherein, "*" represents P < 0.05, and "***" represents P < 0.001.

[0075] The Bifidobacterium adolescentis YG3912 used in the cosmetic composition of the present invention is classified and named Bifidobacterium adolescentis, with a preservation date of June 20, 2024; the preservation unit is the General Microbiology Center of China Microbiological Culture Collection Administration; the preservation address is the Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, China; the preservation number is CGMCC NO.31020.

[0076] The Lactobacillus reuteri YG3708 used in the cosmetic composition of the present invention is classified and named as Limosilactobacillus reuteri, with a preservation date of July 9, 2024; the preservation unit is the General Microbiology Center of the China Microbiological Culture Collection Administration; the preservation address is the Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, China; the preservation number is CGMCC NO.31243. DETAILED DESCRIPTION

[0077] In order to make the purpose, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention. Unless otherwise expressly stated, throughout the specification and claims, the term "including" or its variations such as "comprising" or "including" will be understood to include the stated elements or components, without excluding other elements or other components.

[0078] In addition, in order to better illustrate the present invention, numerous specific details are provided in the specific embodiments below. It should be understood by those skilled in the art that the present invention can also be implemented without certain specific details. In some embodiments, raw materials, elements, methods, means, etc. well known to those skilled in the art are not described in detail, so as to highlight the main purpose of the present invention.

[0079] In the following examples, unless otherwise specified, other reagents or raw materials used are commercially available.

[0080] Example 1: Preparation of fermentation products of Bifidobacterium adolescentis YG3912 and determination of their active ingredients

[0081] The culture medium involved in this embodiment is as follows:

[0082] MRS medium (g / L): peptone 10.0g / L, beef powder 5.0g / L, Tween-80 1mL / L, dipotassium hydrogen phosphate 2.0g / L, yeast powder 4.0g / L, magnesium sulfate 0.2g / L, triammonium citrate 2.0g / L, glucose 20.0g / L, manganese sulfate 0.05g / L, sodium acetate 5.0g / L, L-cysteine ​​hydrochloride 0.5g / L, solvent is water; the preparation method is: mix all the ingredients evenly and sterilize them under high pressure at 121℃ for 15min.

[0083] 1.1 Cultivation of strain YG3912 and preparation of its fermentation broth

[0084] The seed solution of strain YG3912 was transferred to MRS liquid medium at a 2% inoculation rate and cultured anaerobically at 37°C for 24 h. During the culture process, the OD600 value of the culture solution was measured by an ELISA instrument at regular intervals to obtain the growth curve of strain YG3912. Figure 1 shown. Figure 1The results showed that strain YG3912 reached a stable growth phase (OD600=1.22) after 12 hours of culture.

[0085] The fermentation process parameters of strain YG3912 were optimized to obtain the best fermentation parameters as follows: pH 6.25, inoculation volume 2.0%, and culture time 48h. According to the above optimized fermentation parameters, strain YG3912 was fermented to obtain fermentation broth for later use.

[0086] 1.2 Preparation of fermentation filtrate and lysate of strain YG3912

[0087] (1) Preparation of fermentation filtrate (i.e., fermentation supernatant):

[0088] The fermentation broth of strain YG3912 prepared above was centrifuged at 9000 r / min×4°C for 10 min to obtain the fermentation supernatant, which was filtered using a 0.22 μm polyethersulfone filter membrane to obtain the cell-free fermentation filtrate of strain YG3912 (hereinafter referred to as S3912), which was frozen at -80°C for later use.

[0089] (2) Lysate preparation:

[0090] The bacterial cells after centrifugation in the above (1) were harvested, and pre-cooled sterile PBS was added to concentrate them 2 times and resuspend the bacterial cells, and the cell wall was broken by ultrasound. The specific ultrasound program was as follows: ultrasound for 2 seconds, stop for 2 seconds, and the total ultrasound time was 60 minutes. The entire ultrasound program was carried out under ice bath conditions at -4 to 0°C; then, the ultrasound product was heat treated at 80°C for 30 minutes using a high-pressure sterilization equipment to obtain a sterilized lysate of the fermentation broth of strain YG3912 (hereinafter referred to as R3912), which was frozen at -80°C for later use.

[0091] 1.3 Determination of active substances in fermentation filtrate and lysate of strain YG3912

[0092] The nucleic acid and nucleotide content was tested with reference to the group standard "Lysate of Bifida Saccharomyces Fermentation Product as Cosmetic Raw Material", and the oligopeptide and free amino acid content was tested with reference to the national standard GB / T 22729-2008 for marine fish oligopeptide powder.

[0093] (1) Detection of nucleic acid and nucleotide content

[0094] Sample processing: adjust the dilution factor according to the concentration of nucleic acid and nucleotide substances in the sample to be tested so that the absorbance value of the sample at a wavelength of 260nm is between 0.2-0.8.

[0095] Content determination: Using PBS buffer as blank control, take the absorbance of the sample to be tested at a wavelength of 260nm, and calculate the concentration of nucleic acids and nucleotide substances (in terms of nucleic acids) in the sample.

[0096] Result calculation: Reference formula for determination of nucleic acid and nucleotide substances (in terms of nucleic acid) in the sample to be tested:

[0097] c=( s OD260- b OD260)×n×B

[0098] In the above formula:

[0099] c is the concentration of nucleic acid and nucleotide substances (in terms of nucleic acid) in the sample to be tested (μg / mL);

[0100] s OD260 is the absorbance value of the sample under test at a wavelength of 260nm when the optical path is 1cm, and the unit is per centimeter (cm -1 );

[0101] b OD260 is the absorbance value of the blank control at a wavelength of 260nm when the optical path is 1cm, and the unit is per centimeter (cm -1 );

[0102] n is the dilution factor of the sample to be tested;

[0103] B is the average extinction coefficient [(ug / mL) -1 cm -1 ], indicating that 1 unit of absorbance is equivalent to the concentration of total nucleic acid when the optical path length is 1 cm.

[0104] (2) Free amino acids and oligopeptide content

[0105] Weigh 2g (accurate to 0.001g) of the sample to be tested, add 10mL of 15% trichloroacetic acid solution, mix well, and let stand for 10min; centrifuge the sample solution at 4000r / min for 10min, take all the centrifuged supernatant, determine the protein hydrolysate content in the supernatant, and calculate the acid-soluble protein hydrolysate content in the sample to be tested. The protein conversion factor is 6.25.

[0106] The free amino acids were determined by an automatic amino acid analyzer. 0.02 g to 0.03 g of the sample to be tested was weighed accurately to 0.0001 g and uniformly dissolved with a 3.5% sulfosalicylic acid solution. The sample solution to be tested was transferred to a 50 mL volumetric flask and fixed to volume. The sample solution to be tested was centrifuged at a speed of 4000 rpm for 5 min, the supernatant was taken, and the supernatant was filtered with a 0.45 μm microporous filter membrane, the filtrate was transferred to a 50 mL volumetric flask, fixed to volume, and shaken for use. The free amino acid content was determined by an automatic amino acid analyzer.

[0107] The oligopeptide content X1 is calculated according to the following formula:

[0108] X1=X2-X3

[0109] In the above formula:

[0110] X1 represents the oligopeptide content in the sample, in %;

[0111] X2 represents the content of acid-soluble protein hydrolysate in the sample, in %;

[0112] X3 represents the free amino acid content in the sample, in %.

[0113] The results of active substance detection of the lysate of strain YG3912 are shown in Table 1.

[0114] Table 1. Results of determination of active substances in lysate

[0115] serial number Test items unit Test results 1 Nucleic Acids and Nucleotides μg / mL 631.8 2 Oligopeptide mg / kg 6780 3 Free amino acids mg / kg 203

[0116] The results of active substance detection in the fermentation filtrate of strain YG3912 are shown in Table 2.

[0117] Table 2. Results of determination of active substances in fermentation filtrate

[0118]

[0119]

[0120] With reference to the group standard T / GDCA 023-2023 of bifid yeast fermentation product lysate for cosmetic raw materials, the contents of three active ingredients (nucleic acids and nucleotides, oligopeptides and free amino acids) in the fermentation filtrate and lysate were tested. It can be seen from Tables 1 and 2 that the main active substances of both the fermentation filtrate and the lysate of strain YG3912 meet the requirements of the group standard for the content of active ingredients (nucleic acids and nucleotides ≥400.0μg / mL, oligopeptides ≥1000mg / kg, free amino acids ≥50mg / kg).

[0121] Example 2: Evaluation of the effects of Bifidobacterium adolescentis and its fermentation products on regulating skin microecology, soothing, anti-wrinkle and firming

[0122] The culture medium involved in this embodiment is as follows:

[0123] BHI medium (g / L): peptone 10.0g / L, beef heart extract powder 17.5g / L, sodium chloride 5.0g / L, glucose 2.0g / L, disodium hydrogen phosphate (12H2O) 2.5g / L, pH (25℃) = 7.4±0.2, agar (solid addition): 18g / L, solvent is water; its preparation method is: after mixing all the components evenly, sterilize them by high pressure at 121℃ for 15min. When the BHI medium is cooled to 55℃, add 100mL of S3912 supernatant stock solution to 900mL of BHI solid (or liquid) medium at a mass volume ratio of 1:9, and shake and mix to obtain BHI (or liquid) medium containing 10% S3912.

[0124] 2.1 Bacterial culture

[0125] For five strains, including Staphylococcus hominis (ATCC 27844), Corynebacterium simulans (ATCC BAA-15), Moraxella osloensis (ATCC19976), Staphylococcus capitis (ATCC 27840) and Staphylococcus epidermidis (ATCC 12228), 100 μL of the frozen solution was taken and inoculated into BHI liquid medium, and cultured aerobically at 37°C for 24 h until the stable phase; the culture solution in the stable phase was taken and inoculated into new 5 mL of BHI liquid medium at an inoculum volume of 2%, and after aerobic culture at 37°C for 24 h, the concentration of the BHI liquid medium was adjusted to 1×10 7 CFU / mL and reserve for use.

[0126] For four strains including Cutibacterium granulosum (YGMCC 0145), Staphylococcus aureus (ATCC 6538), Propionibacterium acnessubsp.Defendens (ATCC 11828) and Propionibacterium acnessubsp.acnes (ATCC 6919), 500 μL of frozen solution was taken and inoculated into 5 mL of BHI liquid medium, and cultured anaerobically at 37°C for 48 h until the stationary phase; the culture solution at the stationary phase was taken and inoculated into new 5 mL of BHI liquid medium at an inoculum size of 2%, and after anaerobically cultured at 37°C for 48 h, the concentration of 1×10 7CFU / mL is reserved.

[0127] 2.2 In vitro antibacterial experiment

[0128] The paper diffusion method was used to detect the inhibitory effect of 10% S3912 on the above 9 skin bacteria on solid culture medium. 8mL BHI solid culture medium was added to the left side of the two-divided grid as the blank control group (NC group), and 8mL BHI solid culture medium containing 10% (w / v) S3912 was added to the right side as the experimental group (S3912 group). After the culture medium was cooled overnight, sterile filter paper was clamped with sterile tweezers and placed on the agar on both sides. 15μL of bacterial solution from each of the above 9 strains was dripped on the filter paper of the NC group and the S3912 group, respectively. The culture dishes containing S.hominis, C.simulans, M.osloensis, S.capitis and S.epidermidis were aerobically cultured at 37℃ for 24h, while the plates of the other 4 strains were placed in an anaerobic box for 48h, and the diffusion of the bacterial circle was observed. Each group of experiments was repeated three times.

[0129] At the same time, the effect of S3912 on the proliferation of the above 9 strains was detected in BHI liquid culture medium. Specifically, according to the inoculation amount of 2%, the above 9 activated skin bacteria were inoculated into 5mL BHI liquid culture medium (NC group) and 5mL BHI liquid culture medium containing 10% S3912 (S3912 group), respectively. The 15mL centrifuge tubes containing S.hominis, C.simulans, M.osloensis, S.capitis and S.epidermidis were aerobically cultured at 37°C for 24h, and the centrifuge tubes of the other 4 strains were placed in an anaerobic box at 37°C for 48h. After that, the growth rate was measured according to the OD 600 To determine the antibacterial activity of S3912, each group of experiments was repeated three times.

[0130] The results of in vitro antibacterial experiments conducted by the disc diffusion method are shown in Figure 1-4 shown by Figure 1-4 It can be seen that compared with the NC group, S3912 significantly reduced the growth of two pathogens (S. aureus and C. acnes subsp. acnes) on solid culture medium ( Figure 1 ) and M.osloensis ( Figure 2 ) spread area, while six skin resident bacteria including S.hominis, C.simulans, C.granulosum, S.capitis, C.acnes subsp.defendens and S.epidermidis were not intervened ( Figure 3-4 ) of normal diffusion.

[0131] also, Figure 5The effect of S3912 on the proliferation of the above 9 strains in BHI liquid medium is shown; Figure 5 It can be seen that in BHI liquid culture medium, compared with the NC group, S3912 also inhibited the proliferation of two pathogenic bacteria S. aureus (58.61%) and C. acnes subsp. acnes (21.20%) and the aging bacteria M. osloensis (61.30%), while maintaining the normal proliferation of six healthy skin resident bacteria.

[0132] 2.3 Coagglutination experiment

[0133] The activated M.osloensis, S.aureus, C.acnes subsp.acnes and YG3912 inactivated at 80°C for 30 min were centrifuged at 8000×g for 5 min, the supernatant was removed, the precipitate was washed twice with PBS buffer and then resuspended, and the OD600 of the bacterial suspension was adjusted to 1.0±0.1 for later use.

[0134] In a 24-well plate, 300 μL of bacterial suspension of M. osloensis, S. aureus and C. acnes subsp. acnes was added respectively, and then 300 μL of YG3912 (inactivated bacteria) was added respectively. The three groups were cultured in a shaker (150 rpm / min) at 37°C for 12 h. The co-agglutination degree of each group was visually inspected. Each experiment was repeated three times. We referred to the method of Cisar and Rickard (Rickard AH, Leach SA, Hall LS, Buswell CM, High NJ, Handley PS. Phylogenetic relationships and coaggregation ability of freshwater biofilm bacteria. Appl Environ Microbiol. 2002 Jul; 68(7): 3644-50. doi: 10.1128 / AEM.68.7.3644-3650.2002. PMID: 12089055; PMCID: PMC126804) for the visual coaggregation ability scoring system.

[0135] The scoring system was as follows: “-”, no coagglutination occurred; “1+”, a small amount of agglutinate appeared; “2+”, there were visible agglutinates and a small amount of supernatant; “3+”, there were clearly visible agglutinates and supernatant; “4+”, there were a large number of obvious agglutinates and a clearly visible supernatant.

[0136] The results of the coagglutination experiments between the inactivated strain YG3912 and S. aureus, C. acnes subsp. acnes and M. osloensis are shown in Figure 6 ; Figure 6 The results showed that, by visual observation, no coagulation occurred in the NC group (-), the S. aureus group (+++) had clearly visible agglutinates and supernatant, and the C. acnes subsp. acnes group (++) and the M. osloensis group (++) both had visible agglutinates and a small amount of supernatant. The above results show that S3912 can selectively inhibit pathogens while maintaining the proliferation of resident bacteria, thereby maintaining a healthy skin microecology; and by inhibiting the marker bacteria M. osloensis of M-type skin, S3912 can promote the transformation of skin from M-type to C-type, and has great potential to reverse skin aging.

[0137] According to the latest report, there are also two stable community composition patterns of skin microorganisms at the population level in the Chinese population, which are named "skin types" (cutotypes), namely M-cutotype dominated by Moraxella osloensis and C-cutotype dominated by Cutibacterium acnes. The two skin types have different microbial network characteristics, and the microorganisms inside the skin types are positively correlated with each other and negatively correlated with external microorganisms. Metagenomic gene functional enrichment suggests that the C-cutotype skin microbial community is enriched in carbohydrate and sterol metabolism, fatty acid synthesis, etc., while the M-cutotype skin microorganisms are enriched in amino acids, aromatic compound metabolism, drug resistance, etc. Further host clinical manifestations also show obvious differences: C-cutotype skin has a high water and oil content, mostly in young people; while M-cutotype skin has a low water and oil content, a dull and yellowish complexion, and mostly in the elderly.

[0138] Example 3: Evaluation of the skin soothing, anti-wrinkle and skin firming effects of Bifidobacterium adolescentis and its fermentation products

[0139] 3.1 Evaluation of anti-wrinkle function in vitro

[0140] The antioxidant activity of YG3912 fermentation products was evaluated by DPPH free radical scavenging experiment to evaluate its anti-wrinkle function. The experimental method was slightly modified from the method described by Han et al. (Han JY, Kim SK, Lim DW, Kwon O, Choi YR, Kang CH, Lee YJ, Lee YM. Anti-Inflammatory Effect of Ethanol Extract from Hibiscus cannabinus L. Flower in Diesel Particulate Matter-Stimulated HaCaTCells. Nutrients. 2024 Nov 6; 16 (22): 3805. doi: 10.3390 / nu16223805. PMID: 39599592; PMCID: PMC11597620.). DPPH was dissolved in anhydrous ethanol to a final concentration of 100 μM and filtered using a 0.2 μm filter. S3912 and R3912 were diluted with distilled water to prepare experimental group solutions containing 0%, 0.5%, 1.0% and 10.0% of S3912 and R3912. Then, 100 μL of the experimental group solutions were mixed with 100 μL of DPPPH and incubated at 37°C for 30 min. The absorbance was measured at 517 nm using an ELISA reader, and the percentage of antioxidant activity was calculated using the following formula:

[0141] DPPH free radical scavenging rate (%) = 1-(As-A1) / (A C -A0)×100%

[0142] In the above formula, A C is the absorbance of the control group, A0 is the background absorbance of the control group, A S is the absorbance of the sample, and A1 is the absorbance of the sample background.

[0143] 3.2 In vitro firming function evaluation

[0144] The elastase inhibition test is a laboratory test used to evaluate the ability of cosmetic products or ingredients to inhibit the activity of elastase. In this example, the elastin inhibition test was used to evaluate the effect of the YG3912 fermentation product on inhibiting the degradation process of elastin to evaluate its firming function. The specific experimental process is as follows:

[0145] The stock solutions of S3912 and R3912 were diluted with distilled water to prepare 0%, 0.5%, 1.0% and 10.0% S3912 and R3912 as experimental groups. Porcine pancreatic elastase was dissolved in 0.1M Tris-HCl buffer (pH=8.00) to prepare a 10μg / mL working solution. N-succinyl-Ala-Ala-Ala-p-nitroanilide (substrate, AAAPVN) was dissolved in 0.1M Tris-HCl buffer (pH=8.00) to obtain a 1mM AAAPVN solution.

[0146] In addition, 25 μL of 0.1M Tris-HCl buffer, 25 μL of elastase solution and 25 μL of sample or Tris-HCl solution were added to each well of the 96-well plate. The mixture was incubated at 25°C in the dark for 15 min. Then, 100 μL of substrate solution or 100 μL of Tris-HCl buffer was added. After the sample was incubated at 37°C for 15 min, it was read at 530 nm using a microplate reader. The inhibition rate of elastase was calculated using the following formula: Elastase inhibition rate (%) = 1-(As-A1) / (A C -A0)×100%

[0147] In the above formula, A C is the absorbance of the control group, A0 is the background absorbance of the control group, A S is the absorbance of the sample, and A1 is the absorbance of the sample background.

[0148] 3.3 In vitro soothing function evaluation

[0149] In this example, the hyaluronidase inhibition rate of the YG3912 fermentation product was evaluated by referring to the "Guidelines for the Implementation of Alternative Methods for Cosmetic Evaluation" and the "Elson-Morgan Modified Method" to evaluate its skin soothing effect. Specifically, the hyaluronidase inhibition rate test results of the test sample and the negative control were compared. If the hyaluronidase inhibition rate of the test sample is higher than that of the negative control and there is a significant difference (P < 0.05), it can be considered that the test sample has a certain soothing effect.

[0150] Sample configuration:

[0151] (1) 1000 U / mL hyaluronidase solution: Weigh 3.3 mg of hyaluronidase and completely dissolve it in 1.0 mL of acetate buffer solution to obtain a 1000 U / mL enzyme solution, which is prepared before use.

[0152] (2) Acetate buffer solution: 4.8 mL of solution A (0.2 mol / L acetic acid: 11.55 mL of glacial acetic acid dissolved in 1 L of distilled water); 45.2 mL of solution B (0.2 mol / L sodium acetate: 16.4 g of anhydrous sodium acetate or 27.2 g of sodium acetate trihydrate solution dissolved in 1 L of distilled water), mixed and diluted to 100 mL to prepare an acetate buffer solution with a pH of 5.6.

[0153] (3) Acetylacetone solution: Mix 50 mL of 1.0 mol / L sodium carbonate solution and 3.5 mL of acetylacetone solution evenly (prepare before use).

[0154] (4) P-DAB color developer: Accurately weigh 0.8 g of p-dimethylaminobenzaldehyde and dissolve it in 15 mL of concentrated hydrochloric acid and 15 mL of anhydrous ethanol and mix well.

[0155] (5) Calcium chloride solution: 2.5 mol / L; Sodium hydroxide solution: 5 mol / L.

[0156] (6) Vitamin C: 0.6 mg / mL.

[0157] (7) Experimental group: The stock solutions of S3912 and R3912 were diluted with distilled water to prepare 0%, 0.5%, 1.0% and 10.0% S3912 and R3912 as experimental groups.

[0158] Test process:

[0159] The samples were added with various components according to Table 3, where tube A was the sample group, tube B was the sample background group, tube C was the negative control group, and tube D was the negative background group. After the test, 200 μL of the reaction solution was added to a 96-well plate and read at 530 nm using an ELISA reader.

[0160] Table 3. Experimental additive components and procedures

[0161]

[0162] The calculation formula of the hyaluronidase inhibition rate of the test sample is as follows:

[0163] Hyaluronidase inhibition rate (%) = [1-(AB) / (CD)] × 100%

[0164] Among them, A, B, C, and D represent the OD of tube A, tube B, tube C, and tube D respectively. 530 value.

[0165] The test results of DPPH free radical scavenging rate, elastase inhibition rate and hyaluronidase inhibition rate of YG3912 fermentation product are as follows: Figure 7 As shown in Figures A, B, and C.

[0166] Figure 7 Figure A shows that compared with the NC group, the DPPH free radical scavenging rates of the S3912 and R3912 experimental groups at 0.5%, 1.0% and 10% concentrations were significantly different. The DPPH free radical scavenging rates of S3912 and R3912 increased with the increase in concentration, and the 10% S3912 group had the highest free radical scavenging rate of 57.01%. The results show that both S3912 and R3912 have scavenging effects on DPPH free radicals, indicating that S3912 and R3912 have anti-wrinkle effects in vitro.

[0167] Figure 7 Figure B shows that compared with the NC group, the elastase inhibition rates of the S3912 and R3912 experimental groups at 0.5%, 1.0% and 10% concentrations were significantly different. The inhibition rates of S3912 and R3912 on elastase increased with the increase in concentration, with the highest inhibition rate of 39.05% in the 10% R3912 group. This result shows that both S3912 and R3912 have an inhibitory effect on elastase, indicating that both S3912 and R3912 have the effect of tightening the skin.

[0168] Figure 7 Figure C shows that compared with the NC group, the hyaluronidase inhibition rates of the S3912 and R3912 experimental groups at 0.5%, 1.0% and 10% concentrations were significantly different. The inhibition rates of S3912 and R3912 on hyaluronidase increased with the increase in concentration, with the highest inhibition rate of 28.34% in the 10% S3912 group. This result shows that both S3912 and R3912 have an inhibitory effect on hyaluronidase, indicating that both S3912 and R3912 have the effect of soothing the skin in vitro.

[0169] Example 4: Preparation of a bio-based cosmetic composition comprising a YG3912 fermentation product

[0170] In this example, the cosmetic compositions of Preparation Examples 1-4 and Comparative Examples 1-4 were prepared according to the formulations in Table 4 below.

[0171] Table 4. Formulation of cosmetic composition

[0172]

[0173] In the above cosmetic composition formula:

[0174] The fermentation filtrate of Bifidobacterium adolescentis and the fermentation lysate of Bifidobacterium adolescentis are S3912 and R3912 prepared in Example 1, respectively;

[0175] The lactobacillus fermentation product is a fermentation product of Lactobacillus reuteri YG3708 (CGMCC No. 31243), which is obtained by the following method: fermenting and culturing Lactobacillus reuteri YG3708, centrifuging the fermentation broth, collecting the supernatant and filtering it;

[0176] Ginseng root extract, purslane extract, Tremella fuciformis extract and osmanthus extract were all purchased from Bozhou Renzhi Pharmaceutical Co., Ltd., and they are all water extracts of the corresponding Chinese medicinal materials.

[0177] Example 5: Evaluation of the soothing, anti-wrinkle and firming effects of a bio-based skin care composition containing a YG3912 fermentation product (including comparative examples)

[0178] 5.1 Evaluation of anti-wrinkle function in vitro

[0179] The antioxidant activity of the bio-based skin care composition containing the fermentation product of YG3912 was evaluated by DPPH free radical scavenging experiment, and then its anti-wrinkle function was evaluated. The experimental method was the same as that described in "3.1 In vitro anti-wrinkle function evaluation" in Example 3. The experimental group solution was the original solution of Preparation Examples 1-4 and Comparative Examples 1-4 in Example 4.

[0180] Results Figure 8 ;like Figure 8 As shown, the DPPH free radical scavenging ability of the cosmetic compositions of Preparation Examples 1-4 of the present invention is significantly higher than that of the corresponding comparative example groups; in particular, Preparation Example 4 has the strongest scavenging ability, which can reach 55.48%, while Comparative Example 4 basically has no scavenging ability.

[0181] Each preparation example was compared with its corresponding comparative example, the difference being that the latter did not add YG3912 fermentation filtrate. Therefore, the above results indicate that YG3912 fermentation filtrate is a key factor in improving the DPPH free radical scavenging ability of the composition, which may be due to its rich content of short-chain fatty acids, phenolic compounds, amino acids and other bioactive substances. These results emphasize the important role of YG3912 fermentation filtrate in improving anti-wrinkle performance.

[0182] 5.2 In vitro firming function evaluation

[0183] The elastase inhibition rate was used to evaluate the firming function of the bio-based skin care composition containing the YG3912 fermentation product. The experimental method was the same as described in "3.2 In vitro firming function evaluation" in Example 3. The experimental group solution was the original solution of Preparation Examples 1-4 and Comparative Examples 1-4 in Example 4.

[0184] Results Fig. 9 ;like Fig. 9As shown, the elastase inhibition rates of the cosmetic compositions of Preparation Examples 1-4 of the present invention are significantly higher than those of the corresponding comparative examples; in particular, the elastase inhibition rate of Preparation Example 4 is the strongest, reaching 78.21%, while the clearance rate of Comparative Example 4 is only about 10%, a difference of 7.8 times.

[0185] Each preparation example was compared with its corresponding comparative example, the difference being that the latter did not add YG3912 fermentation filtrate. Therefore, the above results indicate that YG3912 fermentation filtrate is a key factor in improving the inhibition rate of elastase, which may be due to its rich content of short-chain fatty acids, phenolic compounds, amino acids and other bioactive substances. These results emphasize the important role of YG3912 fermentation filtrate in improving skin firmness.

[0186] 5.3 In vitro soothing function evaluation

[0187] The soothing function of the bio-based skin care composition containing the YG3912 fermentation product was evaluated by hyaluronidase inhibition rate. The experimental method was the same as described in "3.3 In vitro soothing function evaluation" in Example 3. The experimental group solution was the original solution of Preparation Examples 1-4 and Comparative Examples 1-4 in Example 4.

[0188] Results Fig.10 ;like Fig.10 As shown, the hyaluronidase inhibition rate of the cosmetic compositions of Preparation Examples 1-4 of the present invention is significantly higher than that of the corresponding comparative example group; in particular, the hyaluronidase inhibition rate of Preparation Example 4 is the highest, reaching 54.07%, while the clearance rate of Comparative Example 4 is only 0.98%, a difference of 55 times.

[0189] Each preparation example was compared with its corresponding comparative example, the difference being that the latter did not add YG3912 fermentation filtrate. Therefore, the above results indicate that YG3912 fermentation filtrate is a key factor in improving the hyaluronidase inhibition rate, which may be due to its rich content of short-chain fatty acids, phenolic compounds, amino acids and other bioactive substances. These results emphasize the important role of YG3912 fermentation filtrate in improving soothing ability.

[0190] Example 6: Investigation of the synergistic effect of fermentation products of YG3912 and YG3708

[0191] In this example, the cosmetic compositions of Preparation Example 1, Comparative Example 1 and Comparative Example 2 were prepared according to the formula in Table 5 below, and their in vitro anti-wrinkle function, in vitro firming function and in vitro soothing function were evaluated.

[0192] Table 5. Formulation of cosmetic composition

[0193] Comparative Example 1 Comparative Example 2 Preparation Example 1 Bifidobacterium adolescentis fermentation filtrate / % 0 5 5 Bifidobacterium adolescentis fermentation lysate / % 1 1 1 Lactobacillus fermentation products / % 9.9 0 9.9 Ginseng root extract / % 0.1 0.1 0.1 Portulaca oleracea extract / % 0.5 0.5 0.5 Tremella extract / % 0.5 0.5 0.5 Osmanthus extract / % 0.1 0.1 0.1 water / % 87.9 92.8 82.9

[0194] As shown in Table 5:

[0195] Compared with Preparation Example 1, the difference of Comparative Example 1 is that no Bifidobacterium adolescentis fermentation filtrate is added in Comparative Example 1;

[0196] Compared with Preparation Example 1, the difference of Comparative Example 2 is that no lactobacillus fermentation product is added in Comparative Example 2.

[0197] In the above cosmetic composition formula:

[0198] The fermentation filtrate of Bifidobacterium adolescentis and the fermentation lysate of Bifidobacterium adolescentis are S3912 and R3912 prepared in Example 1, respectively;

[0199] The lactobacillus fermentation product is obtained by the following method: fermenting Lactobacillus reuteri YG3708 (CGMCC No. 31243), centrifuging the fermentation broth, collecting the supernatant and filtering it to obtain;

[0200] Ginseng root extract, purslane extract, Tremella fuciformis extract and osmanthus extract were all purchased from Bozhou Renzhi Pharmaceutical Co., Ltd., and they are all water extracts of the corresponding Chinese medicinal materials.

[0201] 6.1 Evaluation of anti-wrinkle function in vitro

[0202] The DPPH free radical scavenging experiment was used to evaluate the synergistic effect of the fermentation products of YG3912 and YG3708 in terms of anti-wrinkle function. The experimental method was the same as that in Example 3, section "3.1 In vitro anti-wrinkle function evaluation".

[0203] As shown in Table 6, the cosmetic composition of the present invention (i.e., Preparation Example 1) showed significantly higher DPPH free radical scavenging ability compared with Comparative Example 1 and Comparative Example 2. After adding Bifidobacterium adolescentis fermentation filtrate to the formula of Comparative Example 1, the DPPH free radical scavenging rate was significantly increased from 4.44% to 33.52%. Similarly, when lactobacillus fermentation product was added to the formula of Comparative Example 2, the DPPH free radical scavenging rate was also increased from 14.69% to 33.52%.

[0204] From the above results, it can be seen that the addition of Bifidobacterium adolescentis fermentation filtrate and Lactobacillus fermentation product to the cosmetic composition simultaneously produces a synergistic effect in improving the ability of DPPH to scavenge free radicals, rather than a simple superposition effect. This shows that the combined use of Bifidobacterium adolescentis fermentation filtrate and Lactobacillus fermentation product can significantly improve the scavenging rate of DPPH free radicals, thereby achieving a better anti-wrinkle effect.

[0205] Table 6. Anti-wrinkle efficacy evaluation results

[0206]

[0207] 6.2 In vitro firming function evaluation

[0208] The synergistic effect of the fermentation products of YG3912 and YG3708 in terms of firming efficacy was evaluated by using the elastase inhibition rate. The experimental method was the same as that in Example 3, section "3.2 In vitro firming function evaluation".

[0209] As shown in Table 7, the cosmetic composition of the present invention (i.e., Preparation Example 1) to which Bifidobacterium adolescentis fermentation filtrate and Lactobacillus fermentation product were added showed significantly higher elastase inhibition ability compared to Comparative Examples 1 and 2. Table 7 shows that the elastase inhibition rate of Preparation Example 1 is significantly higher than that of Comparative Examples 1 and 2.

[0210] Comparative Examples 1 and 2 lack Bifidobacterium adolescentis fermentation filtrate and Lactobacillus fermentation product, respectively, while Preparation Example 1 contains both. By adding Bifidobacterium adolescentis fermentation filtrate to the formula of Comparative Example 1, the elastase inhibition rate was significantly increased from 7.50% to 15.05%. Similarly, by adding Lactobacillus fermentation product to Comparative Example 2, the elastase inhibition rate was increased from 3.84% to 15.05%. These results show that Bifidobacterium adolescentis fermentation filtrate and Lactobacillus fermentation product have a synergistic effect in improving the elastase inhibition rate, rather than a simple superposition effect. Therefore, it can be concluded that the combination of Bifidobacterium adolescentis fermentation filtrate and Lactobacillus fermentation product can synergistically enhance the elastase inhibition ability of the cosmetic formula, thereby achieving a better firming effect.

[0211] Table 7. Evaluation results of firming effect

[0212]

[0213] 6.3 In vitro soothing function evaluation

[0214] The hyaluronidase inhibition rate was used to evaluate the synergistic effect of the fermentation products of YG3912 and YG3708 in terms of soothing efficacy. The experimental method was the same as that in Example 3, section "3.3 In vitro soothing function evaluation".

[0215] As shown in Table 8, the cosmetic composition of the present invention (i.e., Preparation Example 1) to which Bifidobacterium adolescentis fermentation filtrate and Lactobacillus fermentation product were added showed significantly higher hyaluronidase inhibition ability compared with Comparative Examples 1 and 2. Table 8 shows that the hyaluronidase inhibition rate of Preparation Example 1 is significantly higher than that of Comparative Examples 1 and 2.

[0216] Comparative Examples 1 and 2 lack Bifidobacterium adolescentis fermentation filtrate and Lactobacillus fermentation product, respectively, while Preparation Example 1 contains both. By adding Bifidobacterium adolescentis fermentation filtrate to the formula of Comparative Example 1, the hyaluronidase inhibition rate was significantly increased from 1.96% to 10.43%. Similarly, by adding Lactobacillus fermentation product to Comparative Example 2, the hyaluronidase inhibition rate was increased from 4.58% to 10.43%. These results show that Bifidobacterium adolescentis fermentation filtrate and Lactobacillus fermentation product have a synergistic effect in improving the hyaluronidase inhibition rate, rather than a simple superposition effect. Therefore, it can be concluded that the combination of Bifidobacterium adolescentis fermentation filtrate and Lactobacillus fermentation product can synergistically enhance the hyaluronidase inhibition ability of the cosmetic formula, thereby achieving a better soothing effect.

[0217] Table 8. Soothing effect evaluation results

[0218]

[0219] Example 7: Investigation of the synergistic effect of fermentation products of YG3912 and YG3708

[0220] In this embodiment, the cosmetic compositions of Preparation Example 1, Comparative Example 1 and Comparative Example 2 were prepared according to the formula in Table 9 below, and their in vitro anti-wrinkle function, in vitro firming function and in vitro soothing function were evaluated.

[0221] Table 9. Formulation of cosmetic composition

[0222] Comparative Example 1 Comparative Example 2 Preparation Example 1 Bifidobacterium adolescentis fermentation product filtrate / % 0 10 10 Bifidobacterium adolescentis fermentation lysate / % 1 1 1 Lactobacillus fermentation products / % 9.9 0 9.9 Ginseng root extract / % 0.1 0.1 0.1 Portulaca oleracea extract / % 0.5 0.5 0.5 Tremella extract / % 0.5 0.5 0.5 Osmanthus extract / % 0.1 0.1 0.1 water / % 87.9 87.8 77.9

[0223] Compared with Preparation Example 1, the difference of Comparative Example 1 is that no Bifidobacterium adolescentis fermentation filtrate is added in Comparative Example 1;

[0224] Compared with Preparation Example 1, the difference of Comparative Example 2 is that no lactobacillus fermentation product is added in Comparative Example 2.

[0225] In the above cosmetic composition formula:

[0226] The fermentation filtrate of Bifidobacterium adolescentis and the fermentation lysate of Bifidobacterium adolescentis are S3912 and R3912 prepared in Example 1, respectively;

[0227] The lactobacillus fermentation product is obtained by the following method: fermenting Lactobacillus reuteri YG3708 (CGMCC No. 31243), centrifuging the fermentation broth, collecting the supernatant and filtering it to obtain;

[0228] Ginseng root extract, purslane extract, Tremella fuciformis extract and osmanthus extract were all purchased from Bozhou Renzhi Pharmaceutical Co., Ltd., and they are all water extracts of the corresponding Chinese medicinal materials.

[0229] 7.1 Evaluation of anti-wrinkle function in vitro

[0230] The DPPH free radical scavenging experiment was used to evaluate the synergistic effect of the fermentation products of YG3912 and YG3708 in terms of anti-wrinkle function. The experimental method was the same as that in Example 3, section "3.1 In vitro anti-wrinkle function evaluation".

[0231] As shown in Table 10, the cosmetic composition of the present invention (i.e., Preparation Example 1) showed significantly higher DPPH free radical scavenging ability compared with Comparative Example 1 and Comparative Example 2. After adding Bifidobacterium adolescentis fermentation filtrate to the formula of Comparative Example 1, the DPPH free radical scavenging rate was significantly increased from 6.41% to 52.38%. Similarly, when lactobacillus fermentation product was added to the formula of Comparative Example 2, the DPPH free radical scavenging rate was also increased from 23.44% to 52.38%.

[0232] From the above results, it can be seen that the addition of Bifidobacterium adolescentis fermentation filtrate and Lactobacillus fermentation product to the cosmetic composition simultaneously produces a synergistic effect in improving the ability of DPPH to scavenge free radicals, rather than a simple superposition effect. This shows that the combined use of Bifidobacterium adolescentis fermentation filtrate and Lactobacillus fermentation product can significantly improve the scavenging rate of DPPH free radicals, thereby achieving a better anti-wrinkle effect.

[0233] Table 10. Evaluation of anti-wrinkle efficacy

[0234]

[0235] 7.2 In vitro firming function evaluation

[0236] The synergistic effect of the fermentation products of YG3912 and YG3708 in terms of firming efficacy was evaluated by using the elastase inhibition rate. The experimental method was the same as that in Example 3, section "3.2 In vitro firming function evaluation".

[0237] As shown in Table 11, the cosmetic composition of the present invention (i.e., Preparation Example 1) to which Bifidobacterium adolescentis fermentation filtrate and Lactobacillus fermentation product were added showed significantly higher elastase inhibition ability compared with Comparative Examples 1 and 2. Table 11 shows that the elastase inhibition rate of Preparation Example 1 is significantly higher than that of Comparative Examples 1 and 2.

[0238] Comparative Examples 1 and 2 lack Bifidobacterium adolescentis fermentation filtrate and Lactobacillus fermentation product, respectively, while Preparation Example 1 contains both. By adding Bifidobacterium adolescentis fermentation filtrate to the formula of Comparative Example 1, the elastase inhibition rate was significantly increased from 9.88% to 26.26%. Similarly, by adding Lactobacillus fermentation product to Comparative Example 2, the inhibition rate was increased from 6.91% to 26.26%. These results show that Bifidobacterium adolescentis fermentation filtrate and Lactobacillus fermentation product have a synergistic effect in improving the elastase inhibition rate, rather than a simple superposition effect. Therefore, it can be concluded that the combination of Bifidobacterium adolescentis fermentation filtrate and Lactobacillus fermentation product can synergistically enhance the elastase inhibition ability of the cosmetic formula, thereby achieving a better firming effect.

[0239] Table 11. Evaluation of firming effect

[0240]

[0241] 7.3 In vitro soothing function evaluation

[0242] The hyaluronidase inhibition rate was used to evaluate the synergistic effect of the fermentation products of YG3912 and YG3708 in terms of soothing efficacy. The experimental method was the same as that in Example 3, section "3.3 In vitro soothing function evaluation".

[0243] As shown in Table 12, the cosmetic composition of the present invention (i.e., Preparation Example 1) to which Bifidobacterium adolescentis fermentation filtrate and Lactobacillus fermentation product were added showed significantly higher hyaluronidase inhibition ability compared with Comparative Examples 1 and 2. Table 12 shows that the hyaluronidase inhibition rate of Preparation Example 12 is significantly higher than that of Comparative Examples 1 and 2.

[0244] Comparative Examples 1 and 2 lack Bifidobacterium adolescentis fermentation filtrate and Lactobacillus fermentation product, respectively, while Preparation Example 1 contains both. By adding Bifidobacterium adolescentis fermentation filtrate to the formula of Comparative Example 1, the hyaluronidase inhibition rate was significantly increased from 2.375% to 20.85%. Similarly, by adding Lactobacillus fermentation product to Comparative Example 2, the hyaluronidase inhibition rate was increased from 5.23% to 20.85%. These results show that Bifidobacterium adolescentis fermentation filtrate and Lactobacillus fermentation product have a synergistic effect in improving the hyaluronidase inhibition rate, rather than a simple superposition effect. Therefore, it can be concluded that the combination of Bifidobacterium adolescentis fermentation filtrate and Lactobacillus fermentation product can synergistically enhance the hyaluronidase inhibition ability of the cosmetic formula, thereby achieving a better soothing effect.

[0245] Table 12. Soothing effect evaluation results

[0246]

[0247] Example 8: Investigation of the synergistic effect of fermentation products of YG3912 and YG3708

[0248] In this embodiment, the cosmetic compositions of Preparation Example 1, Comparative Example 1 and Comparative Example 2 were prepared according to the formulations in Table 13 below, and their in vitro anti-wrinkle function, in vitro firming function and in vitro soothing function were evaluated.

[0249] Table 13. Formulation of cosmetic composition

[0250]

[0251]

[0252] As shown in Table 13:

[0253] Compared with Preparation Example 1, the difference of Comparative Example 1 is that no Bifidobacterium adolescentis fermentation filtrate is added in Comparative Example 1;

[0254] Compared with Preparation Example 1, the difference of Comparative Example 2 is that no lactobacillus fermentation product is added in Comparative Example 2.

[0255] In the above cosmetic composition formula:

[0256] The fermentation filtrate of Bifidobacterium adolescentis and the fermentation lysate of Bifidobacterium adolescentis are S3912 and R3912 prepared in Example 1, respectively;

[0257] The lactobacillus fermentation product is obtained by the following method: fermenting Lactobacillus reuteri YG3708 (CGMCC No. 31243), centrifuging the fermentation broth, collecting the supernatant and filtering it to obtain;

[0258] Ginseng root extract, purslane extract, Tremella fuciformis extract and osmanthus extract were all purchased from Bozhou Renzhi Pharmaceutical Co., Ltd., and they are all water extracts of the corresponding Chinese medicinal materials.

[0259] 8.1 Evaluation of anti-wrinkle function in vitro

[0260] The DPPH free radical scavenging experiment was used to evaluate the synergistic effect of the fermentation products of YG3912 and YG3708 in terms of anti-wrinkle function. The experimental method was the same as that in Example 3, section "3.1 In vitro anti-wrinkle function evaluation".

[0261] As shown in Table 14, the cosmetic composition of the present invention (i.e., Preparation Example 1) showed significantly higher DPPH free radical scavenging ability compared with Comparative Example 1 and Comparative Example 2. After adding the fermentation filtrate of Bifidobacterium adolescentis to the formula of Comparative Example 1, the DPPH free radical scavenging rate was significantly increased from 0.57% to 59.08%. Similarly, when the lactobacillus fermentation product was added to the formula of Comparative Example 2, the DPPH free radical scavenging rate was also increased from 47.41% to 59.08%.

[0262] From the above results, it can be seen that the addition of Bifidobacterium adolescentis fermentation filtrate and Lactobacillus fermentation product to the cosmetic composition simultaneously produces a synergistic effect in improving the ability of DPPH to scavenge free radicals, rather than a simple superposition effect. This shows that the combined use of Bifidobacterium adolescentis fermentation filtrate and Lactobacillus fermentation product can significantly improve the scavenging rate of DPPH free radicals, thereby achieving a better anti-wrinkle effect.

[0263] Table 14. Evaluation of anti-wrinkle efficacy

[0264]

[0265] 8.2 In vitro firming function evaluation

[0266] The synergistic effect of the fermentation products of YG3912 and YG3708 in terms of firming efficacy was evaluated by using the elastase inhibition rate. The experimental method was the same as that in Example 3, section "3.2 In vitro firming function evaluation".

[0267] As shown in Table 15, the cosmetic composition of the present invention (i.e., Preparation Example 1) to which Bifidobacterium adolescentis fermentation filtrate and Lactobacillus fermentation product were added showed significantly higher elastase inhibition ability compared to Comparative Examples 1 and 2. Table 15 shows that the elastase inhibition rate of Preparation Example 1 is significantly higher than that of Comparative Examples 1 and 2.

[0268] Comparative Examples 1 and 2 lack Bifidobacterium adolescentis fermentation filtrate and Lactobacillus fermentation product, respectively, while Preparation Example 1 contains both. By adding Bifidobacterium adolescentis fermentation filtrate to the formula of Comparative Example 1, the elastase inhibition rate was significantly increased from 9.97% to 78.21%. Similarly, by adding Lactobacillus fermentation product to Comparative Example 2, the inhibition rate was increased from 45.37% to 78.21%. These results show that Bifidobacterium adolescentis fermentation filtrate and Lactobacillus fermentation product have a synergistic effect in improving the elastase inhibition rate, rather than a simple superposition effect. Therefore, it can be concluded that the combination of Bifidobacterium adolescentis fermentation filtrate and Lactobacillus fermentation product can synergistically enhance the elastase inhibition ability of the cosmetic formula, thereby achieving a better firming effect.

[0269] Table 15. Evaluation of firming effect

[0270]

[0271] 8.3 In vitro soothing function evaluation

[0272] The hyaluronidase inhibition rate was used to evaluate the synergistic effect of the fermentation products of YG3912 and YG3708 in terms of soothing efficacy. The experimental method was the same as that in Example 3, section "3.3 In vitro soothing function evaluation".

[0273] As shown in Table 16, the cosmetic composition of the present invention (i.e., Preparation Example 1) to which Bifidobacterium adolescentis fermentation filtrate and Lactobacillus fermentation product were added showed significantly higher hyaluronidase inhibition ability compared with Comparative Examples 1 and 2. Table 16 shows that the hyaluronidase inhibition rate of Preparation Example 1 is significantly higher than that of Comparative Examples 1 and 2.

[0274] Comparative Examples 1 and 2 lack Bifidobacterium adolescentis fermentation filtrate and Lactobacillus fermentation product, respectively, while Preparation Example 1 contains both. By adding Bifidobacterium adolescentis fermentation filtrate to the formula of Comparative Example 1, the hyaluronidase inhibition rate was significantly increased from 1.98% to 54.07%. Similarly, by adding Lactobacillus fermentation product to Comparative Example 2, the inhibition rate was increased from 45.55% to 54.07%. These results show that Bifidobacterium adolescentis fermentation filtrate and Lactobacillus fermentation product have a synergistic effect in improving the hyaluronidase inhibition rate, rather than a simple superposition effect. Therefore, it can be concluded that the combination of Bifidobacterium adolescentis fermentation filtrate and Lactobacillus fermentation product can synergistically enhance the hyaluronidase inhibition ability of the cosmetic formula, thereby achieving a better soothing effect.

[0275] Table 16. Evaluation of soothing effect

[0276]

[0277] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or make equivalent replacements for some of the technical features therein. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A cosmetic composition, characterized in that The active ingredients of the cosmetic composition include: (1) The fermentation product of Bifidobacterium adolescentis YG3912; wherein the taxonomic name of Bifidobacterium adolescentis YG3912 is Bifidobacterium adolescentis, which is deposited in the General Microbiological Center of China Microbiological Culture Collection Administration, with a deposit date of June 20, 2024 and a deposit number of CGMCC No.31020; (2) Lactobacillus fermentation products; (3) Ginseng root extract; (4) Purslane extract.

2. The cosmetic composition according to claim 1, characterized in that The fermentation product of Bifidobacterium adolescentis YG3912 is its fermentation supernatant and optionally a lysate extracted from its fermentation product.

3. The cosmetic composition according to claim 2, characterized in that The fermentation supernatant of Bifidobacterium adolescentis YG3912 is prepared by the following method: The Bifidobacterium adolescentis YG3912 is fermented and cultured, the fermentation liquid is centrifuged, and the supernatant is collected and filtered to obtain the product.

4. The cosmetic composition according to claim 2, characterized in that The lysate of Bifidobacterium adolescentis YG3912 is extracted by the following method: The Bifidobacterium adolescentis YG3912 is fermented and cultured, the fermentation broth is centrifuged, the bacteria are collected, the bacteria are resuspended in PBS and the cell walls are broken by ultrasound or enzymatic hydrolysis to obtain a lysate; preferably, the obtained lysate is sterilized.

5. The cosmetic composition according to any one of claims 1 to 4, characterized in that The lactobacillus fermentation product is a fermentation product of Lactobacillus reuteri YG3708 with a preservation number of CGMCC No. 31243. The taxonomic name of Lactobacillus reuteri YG3708 is Lactobacillus reuteri, which is deposited in the General Microbiology Center of China National Microbiological Culture Collection Administration, with a preservation date of July 9, 2024 and a preservation number of CGMCC No. 31243. Preferably, the fermentation product of Lactobacillus reuteri YG3708 is its fermentation supernatant; Further preferably, the fermentation supernatant of Lactobacillus reuteri YG3708 is prepared by the following method: The Lactobacillus reuteri YG3708 is fermented and cultured, the fermentation liquid is centrifuged, and the supernatant is collected and filtered to obtain the product.

6. The cosmetic composition according to any one of claims 1 to 5, characterized in that The cosmetic composition further comprises: (5) Tremella fuciformis extract; (6) Osmanthus extract.

7. The cosmetic composition according to any one of claims 1 to 6, characterized in that The ginseng root extract, purslane extract, tremella extract and osmanthus extract are all water extracts.

8. The cosmetic composition according to any one of claims 1 to 7, characterized in that By weight percentage, the cosmetic composition comprises: 0.1-90% of the fermentation supernatant of Bifidobacterium adolescentis YG3912; 0.1-5% lysate of Bifidobacterium adolescentis YG3912; 0.1-10% lactobacillus fermentation products; 0.05-10% ginseng root extract; 0.1-10% purslane extract; 0.01-1% Tremella fuciformis extract; 0.01-1% osmanthus extract; The remainder is cosmetic base.

9. The cosmetic composition according to claim 8, characterized in that By weight percentage, the cosmetic composition comprises: 1-50% of the fermentation supernatant of Bifidobacterium adolescentis YG3912; 1-5% lysate of Bifidobacterium adolescentis YG3912; 1-10% lactobacillus fermentation products; 0.05-1% ginseng root extract; 0.1-5% purslane extract; 0.1-1% Tremella fuciformis extract; 0.05-1% osmanthus extract; The remainder is cosmetic base.

10. Use of the cosmetic composition according to any one of claims 1 to 9 in preparing skin care products with skin soothing, anti-wrinkle and / or firming effects; Preferably, the skin care product is a toner, lotion, cream, mask, essence, disposable lotion or eye cream.

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