Composition, cosmetic and method for preparing composition

Abstract

The invention provides a composition, a cosmetic and a method for preparing the composition, and the composition comprises 1, 3-propylene glycol, lactic acid and lysate of lactobacillus. The composition disclosed by the invention has high moisture retention, low irritation and low viscosity, and can be applied to cosmetics, so that the cosmetics containing the composition have better moisture retention effect, refreshing skin feeling and low irritation, and the quality of the cosmetics is improved.

Description

Technical Field

[0001] This application relates to the field of biochemical engineering. Specifically, this application relates to compositions, cosmetics, and methods for preparing the compositions. Background Technology

[0002] Dry skin usually stems from damage to the stratum corneum barrier function, which can be caused by two main factors: first, environmental factors, such as low temperature and low humidity causing the skin to lose its normal hydration, and excessive degreasing caused by frequent use of soap and detergents; second, with age or long-term exposure to ultraviolet radiation, the stratum corneum loses moisture, thus leading to dry skin. Therefore, effectively maintaining appropriate skin moisture content is key to keeping skin moisturized, soft, and elastic, as well as preventing skin aging. Summary of the Invention

[0003] This invention aims to at least partially solve one of the technical problems existing in the prior art. To this end, this application proposes a composition, a cosmetic, and a method for preparing the composition. The composition of this application possesses high moisturizing properties, low irritation, and low viscosity, and can be applied to cosmetics, enabling cosmetics containing this composition to have better moisturizing effects, a refreshing feel, and low irritation, thereby improving their quality.

[0004] In one aspect of the invention, this application provides a composition. According to embodiments of this application, the composition comprises: 1,3-propanediol, lactic acid, and a lysate of Lactobacillus.

[0005] In the composition according to the embodiments of this application, 1,3-propanediol, lactic acid and lactobacillus lysate are combined to give the composition both good moisturizing properties and low viscosity. It can be applied to cosmetics, giving cosmetics containing the composition better moisturizing effects and a refreshing feel, thereby improving their quality.

[0006] According to embodiments of this application, the above composition may also have the following additional technical features:

[0007] According to embodiments of this application, the lactobacillus includes Lactobacillus reuteri.

[0008] According to an embodiment of this application, the lactobacillus includes *Limosilactobacillus reuteri*, which was deposited on May 29, 2024, at the Guangdong Provincial Center for Microbial Culture Collection (GDMCC) with accession number GDMCC NO: 64700, located at 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou, Guangdong Province, 510070, China.

[0009] According to embodiments of this application, based on the mass of the composition, the composition comprises:

[0010] 70 to 98 parts by weight of 1,3-propanediol;

[0011] 10 to 20 parts by weight of lactic acid;

[0012] 0.5 to 5 parts by weight of Lactobacillus lysate;

[0013] And / or, the pH value of the composition is 4.2 to 6.3.

[0014] In a second aspect of this application, a cosmetic product is provided. According to an embodiment of this application, the cosmetic product comprises the composition described in the first aspect of this application.

[0015] According to embodiments of this application, the content of the composition is 1% to 30% by mass, based on the total mass of the cosmetic.

[0016] According to embodiments of this application, the cosmetic further comprises at least one of: surfactant, thickener, conditioning agent, pH adjuster, salt, preservative, skin feel conditioner, emulsifier, antioxidant, and water.

[0017] According to embodiments of this application, the surfactant includes at least one of sodium cocoyl glycinate, sodium cocoyl glutamate, potassium cocoyl glycinate, potassium cocoyl glutamate, sodium lauroyl glycinate, sodium lauroyl glutamate, sodium methyl lauroyl taurate, cocamidopropyl hydroxysulfonate betaine, disodium cocoamphodiacetate, dodecanoic acid, tetradecanoic acid, and hexadecanoic acid.

[0018] And / or, the thickener includes at least one of cocamide, acrylic copolymers, acrylate copolymers, and hydroxypropyl methylcellulose;

[0019] And / or, the conditioning agent comprises at least one of polyquaternium salt and ethylene glycol distearate;

[0020] And / or, the pH adjuster includes at least one of citric acid, sodium hydroxide, and potassium hydroxide;

[0021] And / or, the salt includes at least one of sodium chloride and potassium chloride;

[0022] And / or, the preservative includes at least one of potassium sorbate and phenoxyethanol;

[0023] And / or, the skin-feeling conditioning agent includes at least one of cyclohexylsiloxane and polydimethylsiloxane;

[0024] And / or, the emulsifier includes at least one of oleyl alcohol polyether, cetyl PEG / PPG-10 / 1 polydimethylsiloxane, sodium lauryl ester, caprylic / capric triglyceride, polyethylene glycol and polypropylene glycol;

[0025] And / or, the antioxidant includes at least one of zinc stearate, tocopheryl acetate, tea polyphenols, tert-butylhydroxybenzoate, butylated hydroxytoluene, lecithin, vitamin C, and vitamin E.

[0026] According to an embodiment of this application, based on the quality of the cosmetic, the cosmetic comprises:

[0027] 1 to 10 parts by weight of the composition described above;

[0028] 15 to 30 parts by weight of surfactant;

[0029] 1 to 3 parts by weight of thickener;

[0030] 5 to 10 parts by weight of the conditioning agent;

[0031] 0.5 to 4 parts by weight of salt;

[0032] 0.1 to 2 parts by weight of preservative;

[0033] water;

[0034] And / or, the cosmetics include cleansing cosmetics.

[0035] According to an embodiment of this application, based on the quality of the cosmetic, the cosmetic comprises:

[0036] 10 to 30 parts by weight of the composition described above;

[0037] 10 to 20 parts by weight of skin conditioning agent;

[0038] 2 to 5 parts by weight of emulsifier;

[0039] 0.5 to 3 parts by weight of antioxidant;

[0040] 0.5 to 3 parts by weight of pH adjuster;

[0041] 0.5 to 3 parts by weight of salt;

[0042] 0.1 to 2 parts by weight of preservative;

[0043] water;

[0044] And / or, the cosmetic product includes at least one of lotion, cream, eye cream and serum.

[0045] In a third aspect of this application, a method for preparing the composition described in the first aspect of this application is provided. According to an embodiment of this application, the method includes: mixing 1,3-propanediol, lactic acid, and a lysate of Lactobacillus to obtain the composition.

[0046] According to embodiments of this application, a method for obtaining the lysate of the lactobacillus includes:

[0047] Lactobacillus was fermented to obtain a fermentation broth;

[0048] The fermentation broth was centrifuged, and the precipitate was collected.

[0049] The precipitate was lysed to obtain the lysate of the lactobacillus.

[0050] According to embodiments of this application, the method for obtaining the lysate of the lactobacillus further includes:

[0051] The pyrolysis solution obtained from the pyrolysis process is subjected to ultrafiltration, and the filtrate is collected.

[0052] The filtrate was subjected to distillation to obtain the lysate of the lactobacillus.

[0053] According to an embodiment of this application, the ultrafiltration membrane used in the ultrafiltration treatment has a molecular weight cutoff of no more than 1000 Da;

[0054] And / or, the temperature of the distillation process is 60°C to 90°C.

[0055] Additional aspects and advantages of the invention will be set forth in part in the description which follows, and in part will be obvious from the description, or may be learned by practice of the invention. Detailed Implementation

[0056] The embodiments of this application are described in detail below. The embodiments described below are exemplary and are only used to explain this application, and should not be construed as limiting this application.

[0057] It should be noted that the terms "first" and "second" are used for descriptive purposes only and should not be construed as indicating or implying relative importance or implicitly specifying the number of technical features indicated. Therefore, a feature defined as "first" or "second" may explicitly or implicitly include one or more of that feature. Furthermore, in the description of this application, unless otherwise stated, "multiple" means two or more.

[0058] The endpoints and any values ​​of the ranges disclosed in this application are not limited to the precise ranges or values, and these ranges or values ​​should be understood to include values ​​close to these ranges or values. For numerical ranges, the endpoint values ​​of the various ranges, the endpoint values ​​of the various ranges and individual point values, and individual point values ​​can be combined with each other to obtain one or more new numerical ranges, which should be considered as specifically disclosed in this application.

[0059] In this application, the terms "comprising" or "including" are open-ended expressions, meaning they include the content specified in this application but do not exclude other aspects.

[0060] In one aspect of the invention, this application provides a composition. According to embodiments of this application, the composition comprises: 1,3-propanediol, lactic acid, and a lysate of Lactobacillus.

[0061] 1,3-Propanediol, as a multifunctional ingredient, can achieve moisturizing effects by enhancing the skin's moisture retention capacity, while also helping to dissolve skin impurities and facilitating the deep penetration of active ingredients, thereby improving skincare and beauty effects. Furthermore, due to its excellent solubility, 1,3-Propanediol can effectively dissolve other ingredients such as fragrances and preservatives, further enhancing the overall performance and stability of cosmetics.

[0062] Lactic acid can enhance the skin's ability to retain moisture, maintain the skin's softness and elasticity, and also has antibacterial properties, which help prevent and treat skin problems such as acne.

[0063] In cosmetics, to meet higher moisturizing requirements, it is often necessary to increase the amount of these moisturizing ingredients. However, increasing the amount of moisturizing ingredients may lead to an increase in product viscosity, thereby affecting the refreshing feeling and comfort during use.

[0064] The substance released after lactobacilli cells rupture is called "lactobacterial lysate," which is rich in active ingredients such as polypeptides, nucleic acids, polysaccharides, organic acids, vitamins, and amino acids. By combining 1,3-propanediol, lactic acid, and lactobacillus lysate, a composition that provides both excellent moisturizing effects and low viscosity can be obtained. This composition not only provides long-lasting moisturizing effects at low concentrations but also reduces the stickiness that occurs at high concentrations, resulting in a more refreshing feel on the skin and significantly improving the quality of cosmetics containing it.

[0065] According to embodiments of this application, the lactobacillus includes *Lactobacillus reuteri*. The lysate of *Lactobacillus reuteri*, combined with 1,3-propanediol and lactic acid, can further enhance the moisturizing effect of the composition and further reduce its viscosity, resulting in a refreshing feel upon application.

[0066] According to embodiments of this application, the lactobacillus includes *Lactobacillus reuteri*, which was deposited on May 29, 2024, at the Guangdong Provincial Center for Microbial Culture Collection (GDMCC) with accession number GDMCC NO: 64700. The lysate of this *Lactobacillus reuteri*, combined with 1,3-propanediol and lactic acid, can further enhance the moisturizing effect of the composition and further reduce its viscosity, resulting in a refreshing feel upon application. In this application, this *Lactobacillus reuteri* is referred to as "*Lactobacillus reuteri* PDO / LA-3" or "PDO / LA-3".

[0067] The metabolic pathway for microbial production of 1,3-propanediol and lactic acid mainly includes: First, glycerol is converted to 3-hydroxypropanal (3-HPA) by glycerol dehydratase (GDHt) within the microorganism; then, 3-hydroxypropanal is reduced to 1,3-propanediol by 1,3-propanediol oxidoreductase (PDOR) under the action of the reducing coenzyme NADH, releasing NAD. + Meanwhile, glycerol dehydrogenases in microorganisms oxidize 3-hydroxypropanal to lactic acid. The pathways of 3-hydroxypropanal reduction to 1,3-propanediol and 3-hydroxypropanal oxidation to lactic acid are in competition with each other; when the yield of 1,3-propanediol increases, the yield of lactic acid tends to decrease.

[0068] Currently, existing technologies primarily produce 1,3-propanediol through microbial fermentation. Because there is competitive inhibition between the biosynthetic pathways of 1,3-propanediol and lactic acid, researchers typically increase 1,3-propanediol yield by inhibiting lactic acid growth through genetic modification or fermentation process optimization. Few researchers have attempted to co-produce 1,3-propanediol and lactic acid using biosynthesis, and even fewer have attempted to simultaneously increase the yields of both. This represents a significant technological barrier in the field.

[0069] A strain of *Limosilactobacillus reuteri* (GDMCC NO: 64700) capable of simultaneously producing high levels of 1,3-propanediol and lactic acid was obtained through UV-plasma combined mutagenesis. This strain exhibits excellent growth and metabolic capacity, as well as high production capacity of 1,3-propanediol and lactic acid, and strong stability even after multiple subcultures. Furthermore, a mass ratio of 1,3-propanediol and lactic acid of (3–6):1 can be readily obtained, which not only further enhances the moisturizing properties of the composition but also allows for the simultaneous production of 1,3-propanediol, lactic acid, and *Limosilactobacillus reuteri* lysate via fermentation. Cell disruption of the fermentation broth yields a composition containing 1,3-propanediol, lactic acid, and *Limosilactobacillus reuteri*, reducing the need for additional 1,3-propanediol and lactic acid addition and lowering workload and production costs. Therefore, the composition of this application can be obtained by fermentation and crushing of *Lactobacillus reuteri*, or by compounding the lysate obtained by fermentation of *Lactobacillus reuteri* with 1,3-propanediol and lactic acid, or by compounding at least one of 1,3-propanediol and lactic acid with a mixture of *Lactobacillus reuteri* fermented, crushed and centrifuged. The specific choice can be made flexibly according to actual needs.

[0070] According to embodiments of this application, based on the mass of the composition, the composition comprises: 70 to 98 parts by weight of 1,3-propanediol; 10 to 20 parts by weight of lactic acid; and 0.5 to 5 parts by weight of Lactobacillus lysate. In some embodiments, the composition comprises 70, 75, 80, 85, 90, 95, or 98 parts by weight of 1,3-propanediol; the composition comprises 10, 12, 14, 15, 16, 18, or 20 parts by weight of lactic acid; and the composition comprises 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, or 5 parts by weight of Lactobacillus reuteri lysate. Therefore, using the above-mentioned proportions of 1,3-propanediol, lactic acid, and Lactobacillus reuteri lysate can further enhance the moisturizing effect of the composition and further reduce its viscosity, resulting in a refreshing feel on the skin during use. Furthermore, considering that the acidity of lactic acid may cause some skin irritation, by adjusting the amount of lactic acid added to meet the above conditions, it is possible to enhance the product's moisturizing properties and reduce viscosity while minimizing potential skin irritation, making the composition suitable for use by people with various skin types.

[0071] According to embodiments of this application, the pH value of the composition is 4.2 to 6.3. In some embodiments, the pH value of the composition is 4.2, 4.5, 4.8, 5.0, 5.1, 5.5, or 6.0, preferably 4.2 to 5.1. Therefore, compositions having the above-mentioned pH values ​​have low skin irritation and are more suitable for use by people with different skin types.

[0072] According to embodiments of this application, the composition further includes a solvent, said solvent being water. 1,3-propanediol, lactic acid, and the lysate of *Lactobacillus reuteri* are all soluble in water, forming a homogeneous mixture.

[0073] In a second aspect, this application discloses a cosmetic product. According to an embodiment of this application, the cosmetic product comprises the composition described in the first aspect. The cosmetic product of this application possesses excellent moisturizing effects, a refreshing feel, and low irritation, making it suitable for people with different skin types.

[0074] According to embodiments of this application, the content of the composition is 1% to 30% by mass based on the total mass of the cosmetic. In some embodiments, the content of the composition is 1% by mass, 5% by mass, 10% by mass, 15% by mass, 20% by mass, 25% by mass, or 30% by mass based on the total mass of the cosmetic. Thus, cosmetics containing the above composition further have better moisturizing effects, a refreshing feel, and low irritation.

[0075] According to embodiments of this application, the cosmetic further comprises at least one of: surfactant, thickener, conditioning agent, pH adjuster, salt, preservative, skin feel conditioner, emulsifier, antioxidant, and water.

[0076] The addition of surfactants helps form a homogeneous and stable emulsion system in the aqueous and oil phases, enhances the dispersibility of components, and also helps active ingredients penetrate the skin better, thereby improving their efficacy. In some embodiments, the surfactant includes at least one selected from sodium cocoyl glycinate, sodium cocoyl glutamate, potassium cocoyl glycinate, potassium cocoyl glutamate, sodium lauroyl glycinate, sodium lauroyl glutamate, sodium methyl lauroyl taurate, cocamidopropyl hydroxysulfobetaine, disodium cocoamphodiacetate, dodecanoic acid, tetradecanoic acid, and hexadecanoic acid.

[0077] Thickeners can increase the viscosity of a product, improve its stability and feel, and make the product easy to apply and less prone to flowing. In some embodiments, the thickener includes at least one of cocamide, acrylic copolymers, acrylate copolymers, and hydroxypropyl methylcellulose.

[0078] Conditioners can improve the sensory properties of a product, such as smoothness, softness, and silkiness, providing a better user experience. In some embodiments, the conditioner includes at least one of polyquaternium salts and glycol distearate.

[0079] pH adjusters maintain the product within a specific pH range, preserving product stability, preventing microbial growth, and conforming to the physiological pH of the skin. In some embodiments, the pH adjuster includes at least one of citric acid, sodium hydroxide, and potassium hydroxide.

[0080] Salts, acting as thickeners or stabilizers, affect product viscosity and the stability of emulsion systems. In some embodiments, the salt includes at least one of sodium chloride and potassium chloride.

[0081] Preservatives protect products from microbial contamination and extend their shelf life. In some embodiments, the preservative includes at least one of potassium sorbate and phenoxyethanol.

[0082] Skin feel agents can improve how a product feels on the skin, such as providing a cooling, refreshing, moisturizing, or matte finish. In some embodiments, the skin feel agent includes at least one of cyclohexylsiloxane and polydimethylsiloxane.

[0083] Emulsifiers stabilize oil-water mixtures, prevent stratification, and form uniform emulsions or pastes. In some embodiments, the emulsifier includes at least one selected from oleyl alcohol polyether, cetyl PEG / PPG-10 / 1 polydimethylsiloxane, sodium laurylate, caprylic / capric triglyceride, polyethylene glycol, and polypropylene glycol.

[0084] Antioxidants prevent the oxidation and deterioration of oils and active ingredients in a product, maintaining its freshness and efficacy. These antioxidants include at least one of zinc stearate, tocopheryl acetate, tea polyphenols, tert-butylhydroxybenzoate, butylated hydroxytoluene, lecithin, vitamin C, and vitamin E.

[0085] According to an embodiment of this application, based on the mass of the cosmetic, the cosmetic comprises: 1 to 10 parts by weight of the composition; 15 to 30 parts by weight of a surfactant; 1 to 3 parts by weight of a thickener; 5 to 10 parts by weight of a conditioning agent; 0.5 to 4 parts by weight of a salt; 0.1 to 2 parts by weight of a preservative; 0.5 to 3 parts by weight of a pH adjuster; and water. In some embodiments, the cosmetic comprises: 1 part by weight, 2 parts by weight, 4 parts by weight, 5 parts by weight, 8 parts by weight, and 10 parts by weight of the composition; 15 parts by weight, 16 parts by weight, 18 parts by weight, 20 parts by weight, 22 parts by weight, 24 parts by weight, 25 parts by weight, 26 parts by weight, 28 parts by weight, and 30 parts by weight of a surfactant; 0.5 parts by weight, 1 part by weight, 1.5 parts by weight, 2 parts by weight, 2.5 parts by weight, and 3 parts by weight of a pH adjuster; 0.1 parts by weight, 0.5 parts by weight, 1 part by weight, 1.5 parts by weight, and 2 parts by weight of a preservative; and water. Thus, cosmetics containing low amounts of the above-mentioned composition still exhibit good moisturizing effects and low irritation, and possess excellent high and low temperature stability and preservative properties.

[0086] According to embodiments of this application, the cosmetics include cleansing cosmetics. These cosmetics offer superior cleansing effects, are low in irritation, and have strong moisturizing properties, leaving the skin feeling moisturized and less prone to tightness after use.

[0087] According to an embodiment of this application, based on the mass of the cosmetic, the cosmetic comprises: 10 to 30 parts by weight of the composition; 10 to 20 parts by weight of a skin feel conditioner; 2 to 5 parts by weight of an emulsifier; 0.5 to 3 parts by weight of an antioxidant; 0.5 to 3 parts by weight of a pH adjuster; 0.5 to 3 parts by weight of a salt; 0.1 to 2 parts by weight of a preservative; and water. In some embodiments, the cosmetic comprises: 10 parts by weight, 15 parts by weight, 20 parts by weight, 25 parts by weight, and 30 parts by weight of an antioxidant; 0.5 parts by weight, 1 part by weight, 1.5 parts by weight, 2 parts by weight, 2.5 parts by weight, and 3 parts by weight of a pH adjuster; 10 parts by weight, 12 parts by weight, 15 parts by weight, 18 parts by weight, and 20 parts by weight of a skin feel conditioner; 2 parts by weight, 3 parts by weight, 4 parts by weight, and 5 parts by weight of an emulsifier; 0.5 parts by weight, 1 part by weight, 1.5 parts by weight, 2 parts by weight, 2.5 parts by weight, and 3 parts by weight of an antioxidant; 0.5 parts by weight, 1 part by weight, 1.5 parts by weight, 2 parts by weight, and 2 parts by weight of a salt; 0.1 parts by weight, 0.5 parts by weight, 1 part by weight, 1.5 parts by weight, and 2 parts by weight of a preservative and water. Therefore, cosmetics containing the above-mentioned low-content composition have both good moisturizing effect, refreshing skin feel and low irritation, and excellent high and low temperature stability and preservative properties.

[0088] According to embodiments of this application, the cosmetic includes at least one of lotion, face cream, eye cream, and serum. The cosmetic with the above-described high-content composition has better moisturizing effects and a refreshing feel, thereby improving the quality of lotion, face cream, eye cream, and serum.

[0089] It should be noted that the features and advantages described above with respect to the composition of the first aspect of this application also apply to this cosmetic product, and will not be repeated here.

[0090] In a third aspect of this application, a method for preparing the composition described in the first aspect of this application is provided. According to an embodiment of this application, the method includes: mixing 1,3-propanediol, lactic acid, and a lysate of Lactobacillus to obtain the composition. By mixing 1,3-propanediol, lactic acid, and a lysate of Lactobacillus, the resulting composition possesses high moisturizing properties, low irritation, and low viscosity, making it suitable for use in cosmetics. This allows cosmetics containing the composition to exhibit better moisturizing effects, a refreshing feel, and low irritation, thereby improving their quality.

[0091] According to embodiments of this application, a method for obtaining the lysate of the lactobacillus includes: fermenting the lactobacillus to obtain a fermentation broth; centrifuging the fermentation broth to collect the precipitate; and lysing the precipitate to obtain the lysate of the lactobacillus. By centrifuging the fermentation broth, the extracellular fluid (supernatant) and cell precipitate can be separated. The cell precipitate is then lysed to rupture the cell membrane and release the internal lysate. When this lysate is combined with 1,3-propanediol and lactic acid, the composition can possess good moisturizing properties, low irritation, and low viscosity.

[0092] According to an embodiment of this application, the method for obtaining the lactobacillus lysate further includes: ultrafiltration of the lysate obtained from the lysis treatment, collecting the filtrate; and distilling the filtrate to obtain the lactobacillus lysate. This further improves the purity, moisture retention, and low irritation of the lysate.

[0093] According to embodiments of this application, the ultrafiltration membrane used in the ultrafiltration process has a molecular weight cutoff of no more than 1000 Da. Therefore, large molecules are retained, while components with moisturizing and low-irritant properties flow out through the ultrafiltration membrane, thereby improving the purity of the lysate and enhancing its moisturizing properties.

[0094] According to embodiments of this application, the distillation temperature is 60°C to 90°C, for example, 60°C, 65°C, 70°C, 75°C, 80°C, 85°C, 90°C, etc. This is to further improve the purity and yield of the moisturizing and low-irritant components in the lysate. The distillation time can be calculated based on the volume of the feed liquid to be distilled; for example, the distillation time for every 200 mL of feed liquid to be distilled is 50 to 70 minutes.

[0095] According to embodiments of this application, the fermentation treatment satisfies at least one of the following conditions:

[0096] The fermentation medium includes at least one of glucose, glycerol, peptone, beef extract, yeast extract, dipotassium hydrogen phosphate, sodium acetate, diammonium hydrogen citrate, magnesium sulfate, manganese sulfate, and Tween-80, along with water. This provides a suitable environment for bacterial growth and metabolism, promoting the production of moisturizing and low-irritant lysates by Lactobacillus.

[0097] The pH of the fermentation medium is 4–6, for example, 4, 4.5, 5, 5.5, 6, etc. This is suitable for bacterial growth and metabolism, and promotes the production of lysates by Lactobacillus that have moisturizing and low-irritant properties.

[0098] The fermentation temperature is 35℃~40℃, for example, it can be 35℃, 36℃, 37℃, 38℃, 39℃, 40℃, etc.;

[0099] The fermentation speed ranges from 100 rpm to 500 rpm, for example, 100 rpm, 200 rpm, 300 rpm, 400 rpm, 500 rpm, etc.

[0100] The fermentation time is 36h to 60h, for example, 36h, 42h, 48h, 54h, 60h, etc.

[0101] During the fermentation process, the culture medium is replenished every 10 to 14 hours. The replenished culture medium includes the initial culture medium and does not contain glycerol.

[0102] Glucose and / or glycerol are added separately during the fermentation process.

[0103] The above fermentation conditions are suitable for cell growth and metabolism, increasing the yield of lysates with high moisture retention and low irritation.

[0104] It should be noted that this application does not impose strict limitations on the fermentation culture container or device; it can be cultured in either a culture flask or a fermenter, and the specific choice can be made flexibly according to the actual situation.

[0105] It is understood that the fermentation culture in this application is actually an expanded fermentation culture. Before fermentation, the Lactobacillus is activated in advance. The activation culture conditions are not strictly limited and can be the conventional activation culture conditions for Lactobacillus in the field, selected flexibly according to the actual situation. In addition, this application does not strictly limit the number of expanded fermentation cultures. It can be carried out only once, or multiple expanded fermentation cultures can be carried out to further increase the bacterial concentration and activity. Each expanded fermentation culture involves inoculating the culture broth obtained from the previous culture into fresh culture medium, and the feeding treatment is carried out during the final expanded fermentation culture.

[0106] During fermentation, culture medium is replenished every 10-14 hours. This replenished medium includes the initial medium but does not contain glycerol. This provides the nutrients required for the growth and metabolism of the microorganisms. Additionally, glucose and glycerol can be added during fermentation to further promote microbial growth and metabolism and to produce high yields of lysates with high moisture retention and low irritation.

[0107] It should be noted that the features and advantages described above with respect to the composition in the first aspect of this application also apply to the method for preparing the composition in the third aspect of this application, and will not be repeated here.

[0108] The present invention will be explained below with reference to embodiments. Those skilled in the art will understand that the following embodiments are for illustrative purposes only and should not be considered as limiting the scope of the invention. Where specific techniques or conditions are not specified in the embodiments, they are performed according to the techniques or conditions described in the literature in the field or according to the product instructions. Reagents or instruments whose manufacturers are not specified are all conventional products that can be obtained commercially.

[0109] Example 1

[0110] 1. Prepare Lactobacillus reuteri PDO / LA-3 (GDMCC NO: 64700) lysate according to the following method.

[0111] 1. Inoculate 5 mL of seed culture medium with *Lactobacillus reuteri* PDO / LA-3 and incubate statically at 37°C for 48 h until OD (Organic Difference) is reached. 600nm =3.0.

[0112] Seed liquid culture medium: 10 g / L peptone, 8 g / L beef extract powder, 4 g / L yeast extract powder, 20 g / L glucose, 2 g / L dipotassium hydrogen phosphate, 5 g / L sodium acetate, 2 g / L diammonium hydrogen citrate, 0.2 g / L magnesium sulfate (MgSO4·7H2O), 0.04 g / L manganese sulfate (MnSO4·4H2O), 1 mL / L Tween-80, with the remainder being water. After preparing the solution, the pH was adjusted to 5.5 using dilute hydrochloric acid and ammonia.

[0113] 2. Add initial culture medium containing 20 g / L glycerol and MRS medium to a 2.5 L fermenter. Purge with nitrogen before inoculation to create anaerobic conditions. Then, inoculate with 10% of Lactobacillus reuteri (preservation number GDMCCNO: 64700) PDO / LA-3 for scale-up fermentation. During the fermentation process, monitor the anaerobic culture at 37℃, pH 5.5 (adjusted by dilute hydrochloric acid and ammonia) and 300 rpm for 48 h.

[0114] MRS medium contains the following components: 10 g / L peptone, 8 g / L beef extract, 4 g / L yeast extract, 20 g / L glucose, 2 g / L dipotassium hydrogen phosphate, 5 g / L sodium acetate, 2 g / L diammonium hydrogen citrate, 0.2 g / L magnesium sulfate (MgSO4·7H2O), 0.04 g / L manganese sulfate (MnSO4·4H2O), 1 mL / L Tween-80, and the remainder is water. After preparing the solution, the pH is adjusted to 5.5 using dilute hydrochloric acid and ammonia. The pH adjuster is 0.1 M dilute hydrochloric acid and 28% ammonia.

[0115] 3. Separation and purification

[0116] (1) Centrifuge the mixture in the fermenter at 8000 rpm and 4℃ for 10 min, wash the precipitate with physiological saline and prepare a cell suspension for cell disruption.

[0117] (2) The supernatant of the mixture after the first centrifugation was placed in a centrifuge equipped with an ultrafiltration membrane with a molecular weight cutoff of 1000 Da and centrifuged for 20 min at 10000 rpm and 4℃. The OD of the fermentation filtrate after the second centrifugation and filtration was... 600 =0.05, clear and transparent in color, pH=4.8;

[0118] (3) The fermentation filtrate was concentrated by distillation at 90℃. The distillation time for each 200 mL of fermentation filtrate was 60 minutes to obtain Lactobacillus reuteri lysate.

[0119] 3. Mix 70 parts of 1,3-propanediol, 18 parts of lactic acid and 2 parts of the Lactobacillus reuteri lysate obtained in the previous step to obtain the composition.

[0120] The differences between Examples 2-12 and Comparative Examples 1-5 and Example 1 are shown in Table 1.

[0121] The difference between Example 5 and Example 1 is that Lactobacillus reuteri GDMCC NO: 64700 is replaced with Lactobacillus reuteri ATCC 53608.

[0122] The difference between Example 6 and Example 1 is that Lactobacillus reuteri GDMCC NO: 64700 is replaced with Lactobacillus reuteri ATCC 55730.

[0123] The difference between Comparative Example 1 and Example 1 is that no lactic acid was added.

[0124] The difference between Comparative Example 2 and Example 1 is that 1,3-propanediol was not added.

[0125] The difference between Comparative Example 3 and Example 1 is that no Lactobacillus reuteri lysate was added.

[0126] The difference between Comparative Example 4 and Example 1 is that 1,3-propanediol was replaced with glycerol.

[0127] The difference between Comparative Example 5 and Example 1 is that Lactobacillus reuteri GDMCC NO: 64700 was replaced with Citrobacter freundii ATCC 8090.

[0128] Test Example 1

[0129] The compositions prepared in Examples 1-12 and Comparative Examples 1-5 were subjected to moisturizing tests, irritation tests, and viscosity tests, respectively. The specific test methods are shown below:

[0130] (1) Moisturizing test

[0131] This study selected 20 participants aged 18-65 years (excluding those who are pregnant or breastfeeding); those without serious systemic diseases, immunodeficiency, or autoimmune diseases; those whose test sites had not undergone skin treatments, cosmetic procedures, or other tests that might affect the results; those without active allergic diseases; those without highly sensitive constitutions; those who had not used hormonal drugs or immunosuppressants within the past month; and those whose test sites were not currently or within the past 3 months participating in other clinical trials.

[0132] Skin moisture content testing was conducted on the forearms of the subjects in a room at 20–22°C and 40%–60% relative humidity. Volunteers were instructed not to expose their forearms to water for 3 hours prior to the test and to wipe the flexor surfaces of their forearms with a tissue before the test. Three 3×3cm square test areas were marked on each of the left and right forearms, spaced 1cm apart. The test area was required to be exposed to the testing environment for at least 20 minutes before the test.

[0133] Before applying the sample, the skin moisture content of different areas of the arm was measured using a Corneometer CM825 (Courage+Khazaka, Germany), and the raw values ​​were recorded.

[0134] 100 μL of test sample was transferred directly to the test area on the subject's arm using a pipette, and rubbed in a circular motion 30 times with the fingers (wearing rubber gloves). Four hours after application, the skin moisture content of the test area was measured using a Corneometer CM825, and the moisturizing coefficient W was calculated. Each test sample was tested 10 times on the skin of different subjects, and the W value was calculated using the average value.

[0135] W = (Skin moisture content 4 hours after sample application - Skin moisture content before sample application) / Skin moisture content before sample application × 100%

[0136] (2) Stimulation test

[0137] The concentrations (IC50) of each sample required to inhibit 50% cell growth were determined using cultured cells from rabbit corneas. Skin irritation was assessed according to the following criteria:

[0138] Mild irritation: IC50 not less than 500 ppm;

[0139] Moderate irritation: IC50 not less than 200 ppm, but less than 500 ppm;

[0140] Strong irritant: IC50 below 200ppm.

[0141] (3) Viscosity test

[0142] The viscosity of the samples was measured using a rotational viscometer, with each sample measured three times and the average value taken.

[0143] The results are shown in Table 1. Compared with Comparative Examples 1-5, the compositions prepared in Examples 1-12 exhibited better moisturizing properties, lower irritation, and lower viscosity, with Example 2 showing the best overall performance. In Comparative Example 1, the absence of lactic acid resulted in a higher pH value and reduced moisturizing properties. In Comparative Example 2, the absence of 1,3-propanediol significantly reduced the moisturizing properties of the composition, while also resulting in low viscosity and strong irritation. In Comparative Example 3, the absence of lysate reduced the moisturizing properties of the composition. In Comparative Example 4, replacing 1,3-propanediol with glycerin resulted in a higher viscosity, which could easily cause a sticky feeling on the skin. In Comparative Example 5, the composition formed using lysate of Citrobacter freundii exhibited lower moisturizing properties.

[0144] Table 1

[0145]

[0146] Example 13

[0147] 1. Formula

[0148] The composition consisted of 5 parts of the composition of Example 2, 5 parts of sodium cocoyl glycinate, 5 parts of sodium lauroyl glutamate, 5 parts of cocamidopropyl hydroxysulfonate betaine, 2 parts of acrylate copolymer, 8 parts of ethylene glycol distearate, 2 parts of citric acid, 2 parts of sodium chloride, 1 part of phenoxyethanol, and water.

[0149] 2. Mix the above ingredients to prepare the facial cleanser.

[0150] Example 14

[0151] 1. Formula

[0152] 20 parts of the composition of Example 2, 15 parts of polydimethylsiloxane, 3 parts of cetyl PEG / PPG-10 / 1 polydimethylsiloxane, 2 parts of zinc stearate, 2 parts of citric acid, 2 parts of sodium chloride, 1 part of potassium sorbate and water.

[0153] 2. Mix the above ingredients to prepare the eye cream.

[0154] Test Example 2

[0155] The facial cleanser prepared in Example 13 and the eye cream prepared in Example 14 were subjected to moisturizing tests, irritation tests and viscosity tests, respectively. For specific test methods, please refer to Test Example 1.

[0156] The results are shown in Table 2. It can be seen that the facial cleanser prepared in Example 13 and the eye cream prepared in Example 14 both have good moisturizing properties, viscosity and low irritation, and are of good quality.

[0157] Table 2

[0158] pH Moisturizing factor W / % Viscosity (25℃) / mPa·s Irritating Example 13 4.52 61.46 42.27 weak stimulation Example 14 6.08 95.38 63.74 weak stimulation

[0159] In the description of this specification, the references to terms such as "one embodiment," "some embodiments," "example," "specific example," or "some examples," etc., indicate that a specific feature, structure, material, or characteristic described in connection with that embodiment or example is included in at least one embodiment or example of the present invention. In this specification, the illustrative expressions of the above terms do not necessarily refer to the same embodiment or example. Furthermore, the specific features, structures, materials, or characteristics described may be combined in any suitable manner in one or more embodiments or examples. Moreover, without contradiction, those skilled in the art can combine and integrate the different embodiments or examples described in this specification, as well as the features of different embodiments or examples.

[0160] Although embodiments of the present invention have been shown and described above, it is understood that the above embodiments are exemplary and should not be construed as limiting the present invention. Those skilled in the art can make changes, modifications, substitutions and variations to the above embodiments within the scope of the present invention.

Claims

1. A composition characterized in that, Comprising: 1,3-propanediol, lactic acid, and a lysate of lactobacillus.

2. The composition of claim 1, wherein, The lactobacillus comprises lactobacillus reuteri; Or, the lactobacillus comprises lactobacillus muci lactobacillus reuteri, which was preserved in Guangdong Microbial Culture Collection Center on May 29, 2024, with a preservation number of GDMCC NO: 64700.

3. The composition according to claim 1 or 2, characterized in that, The composition comprises, based on the mass of the composition: 70 parts by weight to 98 parts by weight of 1,3-propanediol; 10 parts by weight to 20 parts by weight of lactic acid; 0.5 parts by weight to 5 parts by weight of a lysate of lactobacillus; And / or, the pH value of the composition is 4.2 to 6.3; And / or, the composition further comprises a solvent, and the solvent comprises water.

4. A cosmetic product, characterized by Comprising: The composition of any one of claims 1-3.

5. The cosmetic product according to claim 4, characterized in that, The content of the composition is 1 mass% to 30 mass% based on the total mass of the cosmetic.

6. The cosmetic product according to claim 4, characterized in that, Further comprising: At least one of a surfactant, a thickening agent, a conditioning agent, a pH adjuster, a salt, a preservative, a skin feel conditioning agent, an emulsifier, an antioxidant, and water.

7. The cosmetic product according to claim 6, characterized in that, The surfactant comprises at least one of sodium cocoyl glycinate, sodium cocoyl glutamate, potassium cocoyl glycinate, potassium cocoyl glutamate, sodium lauroyl glycinate, sodium lauroyl glutamate, sodium methyl lauroyl taurate, cocamidopropyl hydroxysultaine, disodium cocamphodiacetate, dodecanoic acid, tetradecanoic acid, and hexadecanoic acid; And / or, the thickening agent comprises at least one of cocamide, an acrylic copolymer, an acrylate copolymer, and hydroxypropyl methylcellulose; And / or, the conditioning agent comprises at least one of a polyquaternium and ethylene glycol distearate; And / or, the pH adjuster comprises at least one of citric acid, sodium hydroxide, and potassium hydroxide; And / or, the salt comprises at least one of sodium chloride and potassium chloride; And / or, the preservative comprises at least one of potassium sorbate and phenoxyethanol; And / or, the skin feel conditioning agent comprises at least one of cyclohexasiloxane and dimethicone; And / or, the emulsifier comprises at least one of oleth, cetyl PEG / PPG-10 / 1 dimethicone, sodium laurate, caprylic / capric triglyceride, polyethylene glycol, and polypropylene glycol; And / or, the antioxidant comprises at least one of zinc stearate, tocopheryl acetate, tea polyphenol, tert-butyl hydroxy benzoic acid ester, butylated hydroxytoluene, lecithin, vitamin C, and vitamin E.

8. The cosmetic product according to any one of claims 4 to 7, characterized in that, The cosmetic comprises, based on the mass of the cosmetic: 1 part by weight to 10 parts by weight of the composition; 15 parts by weight to 30 parts by weight of a surfactant; 1 part by weight to 3 parts by weight of a thickening agent; 5 parts by weight to 10 parts by weight of a conditioning agent; 0.5 parts by weight to 4 parts by weight of a salt; 0.1 parts by weight to 2 parts by weight of a preservative; 0.5 parts by weight to 3 parts by weight of a pH adjuster; Water; And / or, the cosmetic comprises a cleansing type cosmetic.

9. The cosmetic product according to any one of claims 4 to 7, characterized in that, The cosmetic comprises, based on the mass of the cosmetic: 10 parts by weight to 30 parts by weight of the composition; 10 parts by weight to 20 parts by weight of a skin feel conditioning agent; 2 parts by weight to 5 parts by weight of an emulsifier; 0.5 parts by weight to 3 parts by weight of an antioxidant; 0.5 parts by weight to 3 parts by weight of a pH adjuster; 0.5 parts by weight to 3 parts by weight of a salt; 0.1 parts by weight to 2 parts by weight of a preservative; water; and / or, the cosmetic product comprises at least one of a lotion, a cream, an eye cream, and an essence.

10. A process for the preparation of a composition according to any one of claims 1 to 3, characterized in that, comprising: mixing 1,3-propanediol, lactic acid, and a lysate of the lactobacillus to obtain the composition.

11. The method of claim 10, wherein, The method for obtaining the lysate of the lactobacillus comprises: fermenting the lactobacillus to obtain a fermentation liquor; centrifuging the fermentation liquor to collect a precipitate; lyzing the precipitate to obtain the lysate of the lactobacillus.

12. The method of claim 11, wherein, The method for obtaining the lysate of the lactobacillus further comprises: ultrafiltering the lysate obtained by the lyzing to collect a filtrate; rectifying the filtrate to obtain the lysate of the lactobacillus.

13. The method of claim 12, wherein, The ultrafiltering is performed by using an ultrafiltering membrane with a molecular weight cut-off of no more than 1000 Da; and / or, the rectifying is performed at a temperature of 60℃ to 90℃.

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