Cosmetic composition containing polyol ester

Abstract

To provide a novel cosmetic composition that contains a polyol ester and offers antibiofilm activity and antibacterial activity.SOLUTION: A cosmetic composition for removing biofilms formed by Staphylococcus aureus and / or Cutibacterium acnes or inhibiting its formation comprises a polyol ester represented by formula (I) (where X is an alditol residue, in which one primary hydroxyl group has been removed from an alditol, and one or more hydroxyl groups may be acetylated, the alditol is mannitol or arabitol, and R2 is a linear hydrocarbon group represented by C13H27 or C15H31).SELECTED DRAWING: Figure 2

Description

[Technical field]

[0001] The present invention relates to a cosmetic composition comprising a polyol ester. [Background technology]

[0002] Infection with Cutibacterium acnes and Staphylococcus aureus causes acne, atopic dermatitis, and the like. There is also a correlation between biofilm formation by these bacteria and skin diseases (Non-Patent Document 1). For this reason, it is necessary to prevent skin infection with Cutibacterium acnes and Staphylococcus aureus, but these bacteria have acquired resistance to antibiotics that have been used conventionally. In light of this, there is a demand for new and more effective substances that have biofilm formation inhibitory activity and antibacterial activity.

[0003] Biosurfactants are substances with surface activity produced by living organisms, and have been attracting attention in recent years because they are safer for living organisms and more biodegradable than artificially synthesized surfactants. Among them, various glycolipids have been reported as biosurfactants produced by microorganisms (Non-Patent Document 2). However, no biosurfactants produced by microorganisms have been reported to have the above-mentioned biofilm formation inhibitory activity or antibacterial properties. [Prior art documents] [Patent documents]

[0004] [Patent Document 1] JP 2018-082636 A [Non-patent literature]

[0005] [Non-Patent Document 1] Pharmaceutics “Microbial Biosurfactants in Cosmetic and Personal Skincare Pharmaceutical Formulations” (2020) 12(11) :1099 [Non-Patent Document 2] Applied Microbiology and Biotechnology “Characterization of cytotoxicity and immunomodulatory effects of glycolipid biosurfactants on human keratinocytes” (2023) 107 :137-152 Summary of the Invention [Problem to be solved by the invention]

[0006] As a result of intensive research, the present inventors have newly discovered that a polyol ester produced by the yeast Rhodosporidium paludigenum BS15 strain (Patent Document 1) has anti-biofilm activity and antibacterial activity. Therefore, an object of the present invention is to provide a novel cosmetic composition that contains the polyol ester and has anti-biofilm activity and antibacterial activity. [Means for solving the problem]

[0007] (1) A cosmetic composition according to one embodiment for achieving the above object comprises: The following formula (I): [ka] (In the formula, X is an alditol residue in which one primary hydroxyl group has been removed from an alditol, and one or more hydroxyl groups may be acetylated, and the alditol is mannitol or arabitol; R2 is C 13 H 27 Or C 15 H 31It is a straight chain hydrocarbon group represented by the formula: and is used for removing or inhibiting the formation of biofilms formed by Staphylococcus aureus and / or Propionibacterium acnes. (2) In the cosmetic composition according to another embodiment, it is preferably used to impart antibacterial activity against Staphylococcus aureus and / or Propionibacterium acnes. (3) In the cosmetic composition according to another embodiment, the polyol ester is preferably contained in an amount of at least 0.1% by mass. (4) In order to achieve the above object, one embodiment of the method for purifying a polyol ester comprises the steps of: The compound of the formula (I) below is produced extracellularly by a microorganism: [ka] (In the formula, X is an alditol residue in which one primary hydroxyl group has been removed from an alditol, and one or more hydroxyl groups may be acetylated, and the alditol is mannitol or arabitol; R2 is C 13 H 27 Or C 15 H 31 It is a straight chain hydrocarbon group represented by the formula: The method includes a step of contacting a culture solution containing a polyol ester represented by the formula (I) with an adsorbent. (5) In the method for purifying a polyol ester according to another embodiment, the adsorbent is preferably activated clay and / or activated carbon. Effect of the Invention

[0008] According to the present invention, it is possible to provide a novel cosmetic composition which contains a polyol ester produced by Rhodosporidium paludigenum strain BS15 and has anti-biofilm activity and antibacterial activity. [Brief description of the drawings]

[0009] [Figure 1]FIG. 1 shows a flow diagram of the process for purifying a polyol ester in a production example. [Diagram 2] FIG. 2 is a photograph showing the results of TLC. [Diagram 3] FIG. 3 is a graph showing the results of a biofilm formation inhibition test. [Figure 4] FIG. 4 is a photograph showing the results of the antibacterial activity test. [Diagram 5] FIG. 5 is a graph showing the results of a protein denaturation test. [Figure 6] FIG. 6 is a photograph showing the results of TLC. [Figure 7] FIG. 7 is a graph showing the results of a growth inhibition test of the JCM 2151 strain by PE. [Figure 8] FIG. 8 is a graph showing the results of a growth inhibition test of the JCM 2874 strain by PE. [Figure 9] FIG. 9 is a graph showing the results of a growth inhibition test of the JCM 2151 strain by various concentrations of PE. [Figure 10] FIG. 10 is a graph showing the results of a growth inhibition test of the JCM 2874 strain by various concentrations of PE. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0010] Next, each embodiment of the present invention will be described with reference to the drawings. Note that each embodiment described below does not limit the invention according to the claims, and all of the elements and combinations thereof described in each embodiment are not necessarily essential to the solution of the present invention.

[0011] (definition) In the present specification, the term "polyol ester" refers to a compound in which a primary hydroxyl group of a polyol, preferably an alditol, is ester-bonded to a carboxyl group of a fatty acid, and is preferably a monoester in which one polyol is bonded to one fatty acid. The primary hydroxyl group refers to a hydroxyl group bonded to a carbon atom to which one carbon atom is bonded. In this polyol ester, the other hydroxyl groups (hydroxyl groups that did not contribute to the ester bond with the fatty acid) on the polyol (preferably an alditol) remain as hydroxyl groups, or one or more of them are acetylated. Examples of alditols (sugar alcohols) include tritol, tetritol, pentitol, hexitol, heptitol, octitol, etc. Preferably, the aditol is pentitol (e.g., arabitol, ribitol, xylitol, etc.) or hexitol (e.g., mannitol, sorbitol, galactitol, allitol, etc.). In a preferred embodiment, the alditol is arabitol or mannitol. The fatty acid may be a linear or branched, saturated or unsaturated fatty acid, but is more preferably a linear saturated fatty acid (e.g., palmitic acid or stearic acid). The fatty acid may be a hydroxy fatty acid, such as a 3-hydroxy fatty acid, and the hydroxyl group may be acetylated. Preferably, the fatty acid is a 3-acetoxy fatty acid, more preferably 3-acetoxypalmitic acid or 3-acetoxystearic acid.

[0012] In the present invention, "removal of biofilm" refers to decomposing, removing, etc., a biofilm formed by Staphylococcus aureus and / or Propionibacterium acnes, whether completely or partially. In the present invention, "inhibition of biofilm formation" refers to inhibiting the formation of a biofilm by Staphylococcus aureus and / or Propionibacterium acnes, whether completely or partially, or reducing the growth rate of a formed biofilm.

[0013] In the present invention, "antibacterial activity" refers to having one or more activities such as bactericidal, bacteriostatic, sterilizing, disinfecting, bacteriostatic, and disinfecting activities against Staphylococcus aureus and / or Propionibacterium acnes.

[0014] (Cosmetic composition) In one embodiment of the present invention, the content of the polyol ester in the cosmetic composition is not particularly limited as long as it is an amount that can remove or inhibit the formation of a biofilm of Staphylococcus aureus and / or Propionibacterium acnes on the skin surface and impart antibacterial activity, taking into consideration the administration form and administration method, etc. For example, the content of the polyol ester is preferably 1% by mass or more, more preferably 0.5% by mass or more, and particularly preferably 0.1% by mass or more, based on the total weight of the cosmetic composition. The content of the polyol ester is, for example, 5% by mass or less, 2.5% by mass or less, 2% by mass or less, or 1% by mass or less, based on the total weight of the cosmetic composition.

[0015] The type of the cosmetic composition of the present invention is not particularly limited, but preferred examples include external preparations such as hair cosmetics, basic cosmetics, makeup cosmetics, fragrance cosmetics, and body cosmetics, and can also be used as oral cosmetics, sanitary products, sanitary cotton, wet tissues, etc. The cosmetic composition of the present invention can be produced according to a conventional method. The cosmetic composition of the present invention can also be used as a medicine, quasi-drug, etc. for external use.

[0016] To explain the types of cosmetic compositions of the present invention in more detail, preferred examples of hair cosmetics include shampoos such as oil shampoo, cream shampoo, conditioning shampoo, anti-dandruff shampoo, hair color shampoo, and rinse-integrated shampoo; rinses, treatments, hair packs, hair foam, hair mousse, hair spray, hair mist, hair wax, hair gel, water grease, setting lotions, color lotions, hair tonics, hair liquids, pomades, hair gels, hair blow dryers, split end coats, hair oils, permanent wave agents, straight perm agents, oxidation hair dyes, hair bleach, hair color pre-treatments, hair color after-treatments, perm pre-treatments, perm after-treatments, hair manicures, and hair growth agents.

[0017] Basic cosmetics include facial cleansers such as cleansing foam, washing powder, face washing powder, cleansing cream, cleansing milk, cleansing lotion, cleansing gel, cleansing oil, and cleansing mask; lotions such as softening lotion, astringent lotion, cleansing lotion, and multi-layered lotion; emulsions such as emollient lotion, moisture lotion, milky lotion, nourishing lotion, nourishing milk, skin moisture, moisture emulsion, massage lotion, cleansing lotion, protective emulsion, sun protect, sun protector, UV care milk, sunscreen, makeup lotion, exfoliating smoother, elbow lotion, hand lotion, and body lotion; emollient cream, nutritious cream, and other skin care products. Creams such as cream, nourishing cream, vanishing cream, moisture cream, night cream, massage cream, cleansing cream, makeup cream, base cream, pre-makeup cream, sunscreen cream, suntan cream, hair removal cream, deodorant cream, shaving cream, and exfoliating cream; gels such as cleansing gel and moisture gel; soaps such as cosmetic soap, transparent soap, medicated soap, liquid soap, shaving soap, and synthetic cosmetic soap; packs and masks such as peel-off packs, powder packs, washing packs, oil packs, and cleansing masks; essences such as moisturizing essences, whitening essences, and UV protection essences; and liposome beauty serums and liposome lotions.

[0018] Preferred examples of makeup cosmetics include face powders, foundations, lipsticks, lip glosses, blushers, eyeliners, mascaras, eye shadows, eyebrow pencils, nail enamel, enamel removers, and nail treatments.

[0019] Preferred examples of the aromatic cosmetic include perfume, perfume, parfum, eau de parfum, eau de toilette, eau de cologne, perfume paste, aromatic powder, perfumed soap, body lotion, and bath oil.

[0020] Preferred examples of body cosmetics include body cleansing agents such as body shampoo; deodorant cosmetics such as deodorant lotions, deodorant powders, deodorant sprays, and deodorant sticks; bleaches, depilatories, and hair removers; bath additives; and insect repellents such as insect repellent sprays.

[0021] In addition, as external cosmetics (external preparations), they can be used in the form of ointments, patches, lotions, liniments, liquid application agents, poultices, plasters, tapes, sheets, aerosols, etc. In addition, they can be used as oral cosmetics such as toothpaste and mouthwash, as well as hygiene products, hygiene cotton, wet tissues, etc.

[0022] Preferred formulations of the cosmetic composition of the present invention include emulsion-type cosmetics such as oil-in-water (O / W) type, water-in-oil (W / O) type, W / O / W type, and O / W / O type, oil-based cosmetics, solid cosmetics, liquid cosmetics, paste cosmetics, stick-type cosmetics, volatile oil-type cosmetics, powder cosmetics, jelly-type cosmetics, gel-type cosmetics, paste-type cosmetics, emulsion polymer-type cosmetics, sheet-type cosmetics, mist-type cosmetics, and spray-type cosmetics.

[0023] In another aspect of the invention, a method for purifying a polyol ester is provided.

[0024] (Production method) In the present invention, the "microorganism" is not particularly limited as long as it has the ability to extracellularly secrete and produce the polyol ester, but is preferably a yeast Rhodosporidium paludigenum strain, more preferably Rhodosporidium paludigenum BS15 strain (hereinafter sometimes simply referred to as BS15 strain), 1042 strain, and 1126 strain, as well as derivatives thereof, which have the ability to extracellularly secrete and produce the polyol ester, and particularly preferably Rhodosporidium paludigenum BS15 strain and derivatives thereof, which have the ability to extracellularly secrete and produce the polyol ester. The Rhodosporidium paludigenum BS15 strain was deposited at the National Institute of Technology and Evaluation, Patent Microorganisms Depositary (NPMD) (Room 122, 2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture, Japan) under the accession number NITE P-02292 on July 8, 2016. The Rhodosporidium paludigenum 1042 strain was deposited at the National Institute of Technology and Evaluation, Patent Microorganisms Depositary (NPMD) under the accession number NITE P-02295 on July 14, 2016. The Rhodosporidium paludigenum 1126 strain was deposited at the National Institute of Technology and Evaluation, Patent Microorganisms Depositary (NPMD) on July 14, 2016 under the accession number NITE P-02296.

[0025] The culture medium used to culture the Rhodosporidium pardigenum of the present invention for the purpose of producing polyol esters contains a carbon source (sugar). The carbon source may be selected from the group consisting of, but is not limited to, glucose, trehalose, galactose, adonitol, L-sorbose, cellobiose, xylitol, salicin, D-glucitol, D-ribose, D-mannitol, D-xylose, dulcitol, L-arabinose, raffinose, D-arabinose, D-gluconic acid, inulin, sucrose, maltose, and glycerol. A more preferred example of the carbon source for producing the polyol esters is glucose (typically, D-glucose).

[0026] In addition to a carbon source, the medium usually contains a nitrogen source and inorganic salts, and may further contain vitamins, amino acids, trace elements, etc., as necessary. As inorganic salts, sodium salts, phosphate salts, magnesium salts, calcium salts, iron salts, manganese salts, etc. can be used.

[0027] More preferably, the medium contains, for example, yeast extract. The medium may contain, for example, one or more selected from the group consisting of peptone, potassium dihydrogen phosphate, disodium hydrogen phosphate, magnesium sulfate heptahydrate, calcium chloride hexahydrate, and iron sulfate heptahydrate. In a preferred embodiment, the medium contains at least D-glucose, yeast extract, potassium dihydrogen phosphate, disodium hydrogen phosphate, magnesium sulfate heptahydrate, and calcium chloride hexahydrate. However, the medium composition is not particularly limited as long as it is suitable for culturing microorganisms.

[0028] The culture can be carried out under normal culture conditions for Rhodosporidium palzigenum. For example, Rhodosporidium palzigenum can be cultured at 26° C. to 34° C., preferably 28° C. to 32° C. The pH of the medium for culturing Rhodosporidium palzigenum is preferably 3 to 10.

[0029] For the production of the polyol ester, it is more preferable to culture Rhodosporidium paldigenum in a medium containing glucose as a carbon source. It is also preferable to culture for typically 5 to 20 days, preferably 7 days or more, more preferably 10 days or more, and even more preferably 11 days or more while adding (replenishing) the carbon source so as not to cause an excess or shortage of the carbon source. The carbon source (sugar) may be added at an appropriate timing, but it is preferable to add sugar successively at a timing such that the sugar concentration is 15% or less, more preferably 10% or less, and even more preferably 6% or less or 1% or less until the total amount of sugar added is 70 to 80% by weight of the initial culture liquid. In one embodiment, sugar may be added once every 3 to 48 hours.

[0030] (Extraction method) The method for extracting polyol ester from the culture solution of Rhodosporidium paldigenum is not particularly limited, and water, an organic solvent, etc. may be used. Examples of the organic solvent include one or a mixture of two or more selected from various organic solvents such as lower alcohols or water-containing lower alcohols such as methanol, ethanol, propyl alcohol, isopropyl alcohol, butanol, and isobutanol, polyhydric alcohols or water-containing polyhydric alcohols such as propylene glycol, 1,3-butylene glycol, 1,2-butylene glycol, 1,4-butylene glycol, 1,5-pentanediol, 1,2-pentanediol, 1,3-pentanediol, 1,4-pentanediol, 1,3,5-pentanetriol, glycerin, and polyethylene glycol (molecular weight: 100 to 100,000), acetone, ethyl acetate, diethyl ether, dimethyl ether, ethyl methyl ether, dioxane, acetonitrile, xylene, benzene, chloroform, carbon tetrachloride, phenol, toluene, and hexane. Among the above organic solvents, ethanol and hexane are preferred, with ethanol being particularly preferred.

[0031] (Adsorbent) The present invention includes, for example, a step of contacting a culture solution containing the polyol ester with an adsorbent. The adsorbent used is not particularly limited as long as it can adsorb and purify the polyol ester, but activated clay and / or activated carbon are preferred. When both activated clay and activated carbon are used as adsorbents, they may be used separately to perform the adsorption step twice in any order, or may be used simultaneously to perform the adsorption step once.

[0032] The present invention will be described in more detail with reference to the following examples, but the present invention is not limited to these examples. In the following examples, the unit % of the numerical values ​​showing the amount of each component added means % by mass. EXAMPLES

[0033] [Example 1] (BS15 strain culture) A liquid medium containing 5% glucose (Wako Pure Chemical Industries), 2.5% yeast extract (powder; Difco Laboratories), 0.12% potassium dihydrogen phosphate (Wako Pure Chemical Industries), 0.03% disodium hydrogen phosphate (Wako Pure Chemical Industries), 0.03% magnesium sulfate heptahydrate (Wako Pure Chemical Industries), and 0.01% calcium chloride hexahydrate (Wako Pure Chemical Industries) was prepared and used for lipid production by the BS15 strain. 25 mL of this liquid medium was added to a 300 mL bladed flask (Asahi Glass) and 5 mL was added to a test tube, and sterilized by autoclaving. One platinum loopful of the BS15 strain on the agar plate was inoculated into the 5 mL liquid medium, and the mixture was cultured overnight at 30 °C and 200 rpm in a thermostatic shaker. The entire amount of the culture was then transferred to 25 mL liquid medium, and the main culture was performed at 30 °C and 140 rpm with rotary shaking. Glucose powder was added to the medium in 3 g portions at 24, 48, 72, 96, 120, 168, and 216 hours after the start of the main culture. The culture was completed after 12 days.

[0034] (Rough purification of polyol ester) The entire culture was transferred to a 50 mL plastic tube and centrifuged at 8000×g for 3 minutes. After confirming that the culture was separated into three layers, the lipids at the bottom, the bacteria in the middle, and the culture supernatant at the top, the culture supernatant was removed, and 20 mL of ethanol was added to the bacteria and lipid layers to dissolve the lipids in ethanol. This was centrifuged at 8000×g for 3 minutes to remove the bacteria and collect the ethanol fraction in which the lipids were dissolved. The ethanol fraction was placed in a desiccator and reduced pressure with a vacuum pump to distill off the ethanol. The remaining lipid was used as a crude polyol ester (hereinafter referred to as crude PE) and purified by the purification method described below.

[0035] (Production example) Method for refining crude PE Figure 1 shows a flow diagram of the process for purifying polyol ester in a production example. Equal amounts of water and hexane were added to the crude PE, and the mixture was left to stand, and only the hexane layer was collected (extraction process, S01). Hexane was distilled off from the collected hexane fraction to obtain purified PE (concentration process, S02).

[0036] (Evaluation of the chemical structure of purified PE) In a past test, the inventors analyzed the chemical structure of PE using NMR and MS, and clarified that PE has the chemical structure of the following formula (I) (Patent Document 1). The PE sample used at this time was used as a standard sample, and comparison was made using the purified PE prepared in this example and thin layer chromatography (TLC). Specifically, a 5% ethanol solution of the standard sample and the purified PE prepared in this example was prepared. After the preparation, the standard sample and the 5% ethanol solution of the purified PE were spotted on a silica gel plate (Merck Millipore) for TLC. After the spotting, the mixture was developed under conditions of 15 minutes using a mixed solvent of hexane and ethyl acetate (6:4). After the development, the solvent on the silica gel plate was dried with a dryer. After the drying, the silica gel plate was immersed in a sulfuric acid methanol solution and heated in an oven to detect organic components contained in the PE sample. These results are shown in FIG. 2. [ka] (In the formula, X is an alditol residue in which one primary hydroxyl group has been removed from an alditol, and one or more hydroxyl groups may be acetylated, and the alditol is mannitol or arabitol; R2 is C 13 H 27 Or C 15 H 31 It is a straight chain hydrocarbon group represented by the formula:

[0037] Figure 2 is a photograph showing the results of TLC. As shown in Figure 2, spots were detected at similar positions for the standard sample and the purified PE (this example). From the above, it was suggested that the purified PE prepared in this example has the chemical structure of formula (I). Specifically, it was suggested that the purified PE prepared in this example has the chemical structures of the following formulas (II) to (IV). [ka] [ka] [ka]

[0038] (Test Example 1) Biofilm formation inhibition test (Preparation of PE solution) The purified PE obtained in the manufacturing example was mixed with tri(caprylic acid / capric acid)glyceryl (manufactured by Kokyu Alcohol Kogyo Co., Ltd.) at room temperature and homogenized to prepare a PE solution with a final concentration of purified PE of 5000 μL / mL. Similarly, PE solutions with final concentrations of 2500, 1250, 625, 312, 156, 78, 19.5, or 0 μL / mL were prepared. In addition, in this test example, the reagents were prepared as follows. The medium used was 700 g of purified water, 17.5 g of Nutrient Broth No. 2 (manufactured by Kanto Chemical Co., Ltd.), 10.5 g of agar, and heat sterilized (hereinafter referred to as NB medium). As PBS, one packet of phosphate buffered saline powder (0.01 mol / L, pH 7.2-7.4) (manufactured by Fujifilm Wako Pure Chemical Co., Ltd.) was diluted with 1 L of purified water and heat sterilized.

[0039] (Preparation of Staphylococcus aureus bacterial solution) Staphylococcus aureus ATCC6538 strain, which had been previously cultured, was added to NB medium at approximately 1.0 × 10 7The bacterial suspension was obtained by diluting the bacterial suspension to CFU / mL. 180 μL of the bacterial suspension was seeded in each well of a 96-well plate. 180 μL of PE solution of each concentration was added to each well seeded with the bacterial suspension. A cover (hereinafter referred to as a 96-well cover) with a protrusion that can be inserted into each well of the 96-well plate was attached so as not to contact the side and bottom of each well. Then, Staphylococcus aureus was cultured at 37 ° C for 24 hours. The 96-well cover was removed from the plate, and a new 96-well plate was similarly attached to the plate in which 180 μL of purified PE of each concentration prepared in NB medium was added to each well. Then, Staphylococcus aureus attached to the 96-well cover was cultured at 36 ° C for 48 hours. Similarly, the 96-well cover was removed, and a new 96-well plate was similarly attached to the plate in which 180 μL of purified PE of each concentration prepared in PBS was added to each well. Thereafter, Staphylococcus aureus attached to the 96-well cover was cultured for 24 hours at 36° C. Similarly, the 96-well cover was removed, and the plate was similarly placed on a new 96-well plate with 200 μL of PBS added to each well, and the 96-well cover was washed. This washing process was repeated twice.

[0040] (Measurement of minimum biofilm inhibitory concentration) 200μL of Crystal Violet solution from Biofilm Formation Assay kit (Dojindo) was added to a new 96-well plate, and a 96-well cover was similarly attached and left to stand at room temperature for 30 minutes. The 96-well cover was washed twice by the above method, and a new 96-well plate to which 200μL of ethanol had been added was similarly attached to the 96-well cover. After leaving the plate to stand at room temperature for 15 minutes, the 96-well cover was removed, and the 96-well plate was stirred, and the absorbance at 570nm was measured. The point at which the decrease in absorbance value reached a plateau was determined as the minimum biofilm inhibitory concentrations (hereinafter, abbreviated as MBIC).

[0041] (result) Figure 3 is a graph showing the results of the biofilm formation inhibition test. In Figure 3, the vertical axis of the graph represents the absorbance value, and the horizontal axis represents the final concentration value of purified PE in each sample. As shown in Figure 3, the absorbance reached a plateau when the concentration of purified PE was 0.0625%. Based on this, the MBIC was determined to be 0.0625%.

[0042] (Test Example 2) Antibacterial activity measurement test (Preparation of acne bacteria liquid) A small amount of P. acnes strain NBRC107605 was scraped off from the glycerol stock with a platinum loop and suspended in 10 mL of modified GAM liquid medium (manufactured by Nissui Co., Ltd.). TM The medium was conditioned under anaerobic conditions using Kenki (manufactured by Mitsubishi Gas Chemical Co., Inc.), and static cultivation was carried out at 37°C for 48 hours. After 48 hours, the container was opened to make the condition aerobic, and the precipitated bacterial cells were collected in a 2.0 mL tube. The solution containing the bacterial cells was centrifuged at 10,000 rpm for 1 minute, after which the supernatant was discarded and 1 mL of PBS was added, and the bacterial cells were suspended by pipetting. This operation was repeated three times. After centrifugation at 1,000 rpm for 1 minute, 0.95 mL of the supernatant was taken and transferred to a 1.5 mL tube, and vortexed.

[0043] (Colony count measurement) Purified PE was diluted with Nonion OT-221R (NOF Corporation) to a final concentration of 20% (hereinafter referred to as diluted PE). Diluted PE or Nonion OT-221R (NOF Corporation) (control) was sterilized by filtration using a 0.45 μm filter. After vortexing on a 96-well plate, the P. acnes supernatant was diluted with PBS so that the final absorbance value at 620 nm was equivalent to 0.10, and added. Diluted PE or Nonion OT-221R was further added to a final concentration of 0.00%, 0.01%, 0.10%, and 1.00%, respectively, to make a 100 μL solution. Anaeropack TMThe solution was made anaerobic using a fermentation kit (manufactured by Mitsubishi Gas Chemical Co., Ltd.), and static culture was carried out at 37°C for 48 hours. After 48 hours, the container was opened to make it aerobic, and 5 μL of the solution was dropped onto modified GAM agar medium (manufactured by Nissui Co., Ltd.) while diluting with PBS as necessary. After the dropped culture solution dried, it was washed with Anaeropack. TM The medium was made anaerobic using Kenki (manufactured by Mitsubishi Gas Chemical Co., Inc.), and static culture was performed at 37°C until sufficient colonies were obtained. After confirming the colonies, the container was opened to make the conditions aerobic, and the colonies were photographed.

[0044] (result) Figure 4 is a photograph showing the results of the antibacterial activity test. As shown in Figure 4, diluted PE clearly reduced the number of P. acnes colonies at a final concentration of 0.01% or more compared to the control. Since no reduction in colonies was observed in the control, it was found that diluted PE has antibacterial activity against P. acnes.

[0045] (Test Example 3) Preservative Effectiveness Activity Test (Preparation of standard bacterial mixture) A mixture of Escherichia coli NBRC3972, Pseudomonas aeruginosa NBRC13275, and Staphylococcus aureus NBRC13276 strains was prepared using a 1.7 x 10 8 Each of the bacteria was added at a concentration of 1 / mL to SCDLP medium (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.) and cultured at 30°C for 20 hours. The bacterial solution after culture was used as the standard bacterial mixture. Purified PE was diluted with Nonion OT-221R (manufactured by NOF Corporation) to a final concentration of 0.1% by weight (hereinafter referred to as the sample). 20 g of the sample was placed in a sterile vial and inoculated with 1% by volume (0.2 mL) of the standard bacterial mixture. The inoculated sample was stored at 30°C, and the viable cell count was measured on the 7th, 14th, and 21st days. The viable cell count was measured using the SCDLP agar medium pour method.

[0046] (result) At a final concentration of 0.1%, purified PE reduced the number of colonies of the standard bacterial mixture to less than 10 on day 14. This result suggested that purified PE has antibacterial activity against a variety of bacteria.

[0047] (Test Example 4) Saponification value measurement test The saponification values ​​of the purified PE of the manufacturing example and the nonionic surfactant (polysorbate) were measured according to the test method described in the Quasi-drug Ingredients Standards 2021.

[0048] (result) The saponification values ​​of the purified PE and polysorbate of the Production Examples were measured in Table 1. As shown in Table 1, the saponification value of the polysorbate was 52.0, whereas the saponification values ​​of the purified PE were high, at 137.5 and 139.8.

[0049] [Table 1]

[0050] (Test Example 5) Protein denaturation test The purified PE described in the manufacturing example, sodium laureth sulfate, a mild surfactant, and sodium lauryl sulfate, a highly irritating surfactant, were mixed to prepare 10 mL of a final concentration of 0.313%, 0.625%, 1.25%, 2.5%, or 5%, respectively, and 0.5 g of Zein protein (manufactured by Kobayashi Perfumery Co., Ltd.) was added to obtain a purified PE (or surfactant)-Zein protein-water mixture. The mixture was mixed at room temperature, left to stand for 30 minutes to reach dissolution equilibrium, and each mixture was centrifuged (3000 rpm, 20 minutes) to separate the undissolved Zein protein into solid and liquid. The sediment was washed with purified water to remove the purified PE (or surfactant) residue, and vacuum dried for 48 hours. After the vacuum drying, the mixture was further dried at 105°C for 48 hours to obtain the weight of undissolved Zein. The Zein protein denaturation rate was calculated by the following formula. Zein protein denaturation rate (%) = [(0.5 g - weight of undissolved zein (g)) / 0.5 g] x 100

[0051] (result) FIG. 5 is a graph showing the results of the protein denaturation test. In FIG. 5, the vertical axis of the graph shows the denaturation rate of Zein protein, and the horizontal axis shows the final concentration after addition of each sample. As shown in FIG. 5, at a final concentration of 5%, the protein denaturation rate of purified PE was 59.32%, and the protein denaturation rate of sodium laureth sulfate was 57.44%. In the experiment using sodium lauryl sulfate, the denaturation rate was 100% at a final concentration of 2.5%. Since the protein denaturation rates of sodium laureth sulfate, a low-irritation surfactant, and purified PE were similar, it is considered that purified PE is less irritating to proteins.

[0052] [Example 2] (BS15 strain culture) A liquid medium containing 5% glucose (Wako Pure Chemical Industries), 1.5% yeast extract (powder; Difco Laboratories), 0.12% potassium dihydrogen phosphate (Wako Pure Chemical Industries), 0.03% disodium hydrogen phosphate (Wako Pure Chemical Industries), 0.03% magnesium sulfate heptahydrate (Wako Pure Chemical Industries), 0.02% calcium chloride hexahydrate (Wako Pure Chemical Industries), and 0.005% ferrous sulfate heptahydrate (Wako Pure Chemical Industries) was prepared. 60 mL of this liquid medium was placed in a 500 mL bladed flask (Asahi Glass) and sterilized in an autoclave. After the sterilization, the BS15 strain on the agar plate was inoculated into this liquid medium, and pre-cultured for 2 days at 28 ° C. and 120 rpm using a thermostatic shaker. After that, 540 mL of the same liquid medium as in the pre-culture was added to a 2 L Bioneer jar fermenter (Marubishi Bioengine) and sterilized by autoclaving, and the entire amount of the culture medium obtained in the pre-culture was added to start the main culture. In the main culture, the culture medium was maintained at 28°C and stirred at 600 rpm while aerating the culture tank with air at 1 L per minute. The main culture was carried out for 168 hours, during which time approximately 800 mL of 50% glucose solution was gradually added using a pump.

[0053] (Purification of polyol ester) The entire amount of the culture solution obtained in the main culture was transferred to a beaker and left to stand until the polyol ester precipitated at the bottom of the beaker. After the standing, 300 mL of chloroform was added and gently stirred to dissolve the polyol ester in the chloroform layer. After the dissolution, the chloroform layer was collected with a glass pipette and transferred to an eggplant-shaped flask. Then, the chloroform was distilled off using a rotary evaporator (Tokyo Rikakikai), and the remaining polyol ester was collected. This was used as purified PE.

[0054] (Evaluation of the chemical structure of purified PE) In a past test, the inventors analyzed the chemical structure of PE using NMR and MS, and clarified that PE has the chemical structure of the following formula (I) (Patent Document 1). The PE sample used at that time was used as a standard sample, and comparison was made using the purified PE prepared in this example and thin layer chromatography (TLC). Specifically, a 5% ethanol solution of the standard sample and the purified PE prepared in this example was prepared. After the preparation, the standard sample and the 5% ethanol solution of the purified PE were spotted on a silica gel plate (Merck Millipore) for TLC. After the spotting, the mixture was developed under conditions of 15 minutes using a mixed solvent of hexane and ethyl acetate (6:4). After the development, the solvent on the silica gel plate was dried with a dryer. After the drying, the silica gel plate was immersed in a sulfuric acid methanol solution and heated in an oven to detect organic components contained in the PE sample. These results are shown in FIG. 6. [ka] (In the formula, X is an alditol residue in which one primary hydroxyl group has been removed from an alditol, and one or more hydroxyl groups may be acetylated, and the alditol is mannitol or arabitol; R2 is C 13 H 27 Or C 15 H 31 It is a straight chain hydrocarbon group represented by the formula:

[0055] Figure 6 is a photograph showing the results of TLC. As shown in Figure 6, the standard sample and the purified PE (this example) had spots detected at similar positions, suggesting that the purified PE prepared in this example has the chemical structure of formula (I). This suggests that the PE prepared in this example has the chemical structures of formulas (II) to (IV) similar to the PE prepared in Example 1.

[0056] [Example 3] (Staphylococcus aureus culture) An LB liquid medium containing 1.0% (w / v) peptone, 1.0% (w / v) sodium chloride, and 0.5% (w / v) yeast extract was prepared for use in culturing Staphylococcus aureus. 5.0 mL of the LB liquid medium was dispensed into a test tube and sterilized by autoclaving. After the sterilization, a small amount of Staphylococcus aureus was scraped from the glycerol stock with a P200 tip and inoculated into the LB liquid medium. After the inoculation, the medium was cultured at 37°C for 16 hours using a thermostatic shaker. After the culture, the Staphylococcus aureus culture solution was used for each test. The Staphylococcus aureus strains used were JCM 2151 and JCM 2874.

[0057] (Test Example 1) Growth Inhibition Test 1 The crudely purified PE, which was roughly purified by the same method as in Example 1, was diluted with LB liquid medium to a final concentration of 1.0% (w / v). After the dilution, the diluted crudely purified PE (hereinafter also referred to as "diluted PE") was used to prepare PE-containing LB liquid medium with a final concentration of 0% (w / v), 0.5% (w / v), 0.25% (w / v), 0.125% (w / v), 0.0625% (w / v), and 0.03125% (w / v). After the adjustment, 5 mL of the PE-containing LB liquid medium at each concentration was dispensed into an L-shaped test tube that had been sterilized by autoclaving and then dry-heat-treated. After the dispensing, the Staphylococcus aureus culture solution was seeded at a concentration of 1% (v / v). After the seeding, the medium was cultured at 37°C and 70 rpm using a small shaking culture device (ADVANTEC, TVS062CA). In the above culture, the absorbance at 660 nm (OD660) at the start of the culture was set to 0, and OD660 was measured every 30 minutes. These results are shown in Figures 7 and 8.

[0058] Fig. 7 is a graph showing the results of a test of the growth inhibition of JCM 2151 strain by PE. In Fig. 7, the vertical axis represents the OD660 value, and the horizontal axis represents the culture time (hours). The values ​​on the vertical axis in Figure 7 are as follows: 0% <h2 style=";text-align:left;direction:ltr">0h:0, 0.5h:0.011, 1h:0.087, 1.5h:0.243, 2h:0.483, 2.5h:0.768, 3h:1.108, 3.5h:1.500, 4h:1.880, 4.5h:2.191, 5h:2.374, 5.5h:2.488, 6h:2.560 6.5h:2.609、7h:2.639、7.5h:2.676、8h:2.695、8.5h:2.724、9h:2.735、9.5h:2.760、10h:2.778、10.5h:2.790、11h:2.789、11.5h:2.787、12h:2.807、1 2.5h:2.813, 13h:2.817, 13.5h:2.828, 14h:2.835, 14.5h:2.843, 15h:2.861, 15.5h:2.844, 16h:2.849, 16.5h:2.860, 17h:2.862, 17.5h:2.872, 18h:2 .884, 18.5h:2.879, 19h:2.874, 19.5h:2.886, 20h:2.919, 20.5h:2.900, 21h:2.908, 21.5h:2.900, 22h:2.906, 22.5h:2.913, 23h:2.908, 23.5h:2.930<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> 0.03125%<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr">0h:0, 0.5h:0.004, 1h:0.055, 1.5h:0.228, 2h:0.488, 2.5h:0.798, 3h:1.178, 3.5h:1.593, 4h:1.948, 4.5h:2.207, 5h:2.384, 5.5h:2.500, 6h:2.554 6.5h:2.604, 7h:2.625, 7.5h:2.648, 8h:2.686, 8.5h:2.702, 9h:2.727, 9.5h:2.750, 10h:2.772, 10.5h:2.777, 11h:2.784, 11.5h:2.791, 12h:2.794, 1 2.5h:2.808, 13h:2.808, 13.5h:2.810, 14h:2.823, 14.5h:2.816, 15h:2.817, 15.5h:2.824, 16h:2.836, 16.5h:2.827, 17h:2.836, 17.5h:2.859, 18h:2 .861, 18.5h:2.852, 19h:2.858, 19.5h:2.862, 20h:2.882, 20.5h:2.873, 21h:2.888, 21.5h:2.883, 22h:2.890, 22.5h:2.880, 23h:2.897, 23.5h:2.900<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> 0.0625%<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr">0h:0, 0.5h:0.037, 1h:0.035, 1.5h:0.214, 2h:0.483, 2.5h:0.801, 3h:1.183, 3.5h:1.583, 4h:1.915, 4.5h:2.144, 5h:2.297, 5.5h:2.389, 6h:2.452 6.5h:2.508, 7h:2.541, 7.5h:2.543, 8h:2.563, 8.5h:2.601, 9h:2.640, 9.5h:2.637, 10h:2.637, 10.5h:2.668, 11h:2.666, 11.5h:2.652, 12h:2.652, 1 2.5h:2.656, 13h:2.672, 13.5h:2.676, 14h:2.662, 14.5h:2.679, 15h:2.687, 15.5h:2.682, 16h:2.700, 16.5h:2.708, 17h:2.715, 17.5h:2.701, 18h:2 .713, 18.5h:2.718, 19h:2.726, 19.5h:2.734, 20h:2.736, 20.5h:2.744, 21h:2.743, 21.5h:2.754, 22h:2.751, 22.5h:2.771, 23h:2.759, 23.5h:2.766<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> 0.125%<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr">0h:0, 0.5h:0.171, 1h:-0.001, 1.5h:0.154, 2h:0.398, 2.5h:0.696, 3h:1.065, 3.5h:1.479, 4h:1.810, 4.5h:2.027, 5h:2.185, 5.5h:2.257, 6h:2.289 6.5h:2.347、7h:2.371、7.5h:2.382、8h:2.411、8.5h:2.439、9h:2.460、9.5h:2.460、10h:2.475、10.5h:2.477、11h:2.470、11.5h:2.478、12h:2.476、1 2.5h:2.462, 13h:2.472, 13.5h:2.474, 14h:2.478, 14.5h:2.484, 15h:2.470, 15.5h:2.489, 16h:2.492, 16.5h:2.499, 17h:2.495, 17.5h:2.507, 18h:2 .504, 18.5h:2.513, 19h:2.531, 19.5h:2.538, 20h:2.543, 20.5h:2.553, 21h:2.538, 21.5h:2.564, 22h:2.562, 22.5h:2.568, 23h:2.577, 23.5h:2.581<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> 0.25%<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr">0h:0, 0.5h:0.310, 1h:-0.032, 1.5h:0.118, 2h:0.362, 2.5h:0.648, 3h:1.000, 3.5h:1.442, 4h:1.771, 4.5h:2.005, 5h:2.186, 5.5h:2.288, 6h:2.353 6.5h:2.391, 7h:2.419, 7.5h:2.438, 8h:2.469, 8.5h:2.483, 9h:2.504, 9.5h:2.503, 10h:2.506, 10.5h:2.496, 11h:2.489, 11.5h:2.476, 12h:2.481, 1 2.5h:2.472、13h:2.471、13.5h:2.458、14h:2.461、14.5h:2.448、15h:2.447、15.5h:2.451、16h:2.446、16.5h:2.447、17h:2.447、17.5h:2.452、18h:2 .458、18.5h:2.463、19h:2.472、19.5h:2.475、20h:2.496、20.5h:2.494、21h:2.502、21.5h:2.509、22h:2.514、22.5h:2.520、23h:2.536、23.5h:2.534<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> 0.5%<h2 style=";text-align:left;direction:ltr"> 0h:0, 0.5h:0.421, 1h:0.097, 1.5h:0.236, 2h:0.417, 2.5h:0.631, 3h:0.8 87, 3.5h:1.192, 4h:1.432, 4.5h:1.569, 5h:1.666, 5.5h:1.739, 6h:1.775, 6.5h:1.789, 7h:1.825, 7.5h:1.836, 8h:1.851, 8.5h:1.866, 9h:1.878, 9.5 h:1.881, 10h:1.889, 10.5h:1.894, 11h:1.884, 11.5h:1.892, 12h:1.885, 1 2.5h:1.899, 13h:1.885, 13.5h:1.890, 14h:1.896, 14.5h:1.886, 15h:1.88 9, 15.5h:1.877, 16h:1.877, 16.5h:1.880, 17h:1.880, 17.5h:1.872, 18h:1 .874, 18.5h:1.874, 19h:1.890, 19.5h:1.882, 20h:1.887, 20.5h:1.897, 21 h:1.913, 21.5h:1.909, 22h:1.912, 22.5h:1.924, 23h:1.921, 23.5h:1.922 As shown in FIG. 7, it was found that the OD660 value decreased depending on the concentration of PE contained in the LB liquid medium.

[0059] Fig. 8 is a graph showing the results of a test of the growth inhibition of JCM 2874 strain by PE. In Fig. 8, the vertical axis represents the OD660 value, and the horizontal axis represents the culture time (hours). The values ​​on the vertical axis in Figure 8 are as follows: 0% <h2 style=";text-align:left;direction:ltr">0h:0, 0.5h:0.018, 1h:0.089, 1.5h:0.264, 2h:0.554, 2.5h:0.911, 3h:1.370, 3.5h:1.845, 4h:2.248, 4.5h:2.546, 5h:2.737, 5.5h:2.856, 6h:2.936 6.5h:2.923, 7h:2.969, 7.5h:2.956, 8h:2.998, 8.5h:2.997, 9h:2.991, 9.5h:3.005, 10h:3.013, 10.5h:2.990, 11h:2.997, 11.5h:2.983, 12h:2.984, 1 2.5h:2.973, 13h:2.956, 13.5h:2.943, 14h:2.955, 14.5h:2.956, 15h:2.940, 15.5h:2.938, 16h:2.933, 16.5h:2.944, 17h:2.913, 17.5h:2.916, 18h:2 .918, 18.5h:2.908, 19h:2.882, 19.5h:2.897, 20h:2.898, 20.5h:2.882, 21h:2.879, 21.5h:2.883, 22h:2.867, 22.5h:2.870, 23h:2.853, 23.5h:2.868<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> 0.03125%<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr">0h:0, 0.5h:-0.016, 1h:0.044, 1.5h:0.216, 2h:0.527, 2.5h:0.902, 3h:1.409, 3.5h:1.931, 4h:2.326, 4.5h:2.642, 5h:2.870, 5.5h:3.017, 6h:3.108 6.5h:3.178、7h:3.192、7.5h:3.208、8h:3.225、8.5h:3.229、9h:3.238、9.5h:3.250、10h:3.239、10.5h:3.225、11h:3.233、11.5h:3.232、12h:3.199、1 2.5h:3.208、13h:3.202、13.5h:3.190、14h:3.175、14.5h:3.179、15h:3.178、15.5h:3.160、16h:3.145、16.5h:3.132、17h:3.130、17.5h:3.125、18h:3 .127、18.5h:3.102、19h:3.123、19.5h:3.103、20h:3.100、20.5h:3.089、21h:3.069、21.5h:3.086、22h:3.079、22.5h:3.070、23h:3.059、23.5h:3.058<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> 0.0625%<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr">0h:0, 0.5h:-0.050, 1h:0.001, 1.5h:0.172, 2h:0.482, 2.5h:0.854, 3h:1.372, 3.5h:1.926, 4h:2.331, 4.5h:2.635, 5h:2.823, 5.5h:2.980, 6h:3.064 6.5h:3.110, 7h:3.118, 7.5h:3.149, 8h:3.152, 8.5h:3.170, 9h:3.156, 9.5h:3.157, 10h:3.148, 10.5h:3.137, 11h:3.140, 11.5h:3.120, 12h:3.093, 1 2.5h:3.106, 13h:3.084, 13.5h:3.083, 14h:3.066, 14.5h:3.066, 15h:3.069, 15.5h:3.052, 16h:3.034, 16.5h:3.047, 17h:3.041, 17.5h:3.023, 18h:3 .008, 18.5h:2.997, 19h:3.000, 19.5h:3.026, 20h:2.988, 20.5h:2.978, 21h:2.983, 21.5h:2.968, 22h:2.967, 22.5h:2.955, 23h:2.960, 23.5h:2.933<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> 0.125%<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr">0h:0, 0.5h:-0.082, 1h:-0.043, 1.5h:0.117, 2h:0.419, 2.5h:0.793, 3h:1.309, 3.5h:1.868, 4h:2.244, 4.5h:2.534, 5h:2.741, 5.5h:2.897, 6h:3.028 6.5h:3.097, 7h:3.145, 7.5h:3.192, 8h:3.206, 8.5h:3.226, 9h:3.218, 9.5h:3.194, 10h:3.189, 10.5h:3.154, 11h:3.131, 11.5h:3.118, 12h:3.087 12.5h:3.085, 13h:3.070, 13.5h:3.046, 14h:3.030, 14.5h:3.025, 15h:3.020, 15.5h:3.012, 16h:2.999, 16.5h:2.982, 17h:2.984, 17.5h:2.975, 18h: 2.967, 18.5h:2.961, 19h:2.956, 19.5h:2.938, 20h:2.935, 20.5h:2.924, 21h:2.919, 21.5h:2.920, 22h:2.916, 22.5h:2.900, 23h:2.906, 23.5h:2.899<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> 0.25%<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr">0h:0, 0.5h:-0.079, 1h:-0.043, 1.5h:0.120, 2h:0.430, 2.5h:0.809, 3h:1.317, 3.5h:1.896, 4h:2.251, 4.5h:2.522, 5h:2.745, 5.5h:2.887, 6h:2.982 6.5h:3.025, 7h:3.059, 7.5h:3.072, 8h:3.069, 8.5h:3.072, 9h:3.040, 9.5h:3.029, 10h:3.019, 10.5h:3.000, 11h:2.976, 11.5h:2.967, 12h:2.950 12.5h:2.940, 13h:2.923, 13.5h:2.916, 14h:2.893, 14.5h:2.880, 15h:2.865, 15.5h:2.856, 16h:2.854, 16.5h:2.850, 17h:2.850, 17.5h:2.851, 18h: 2.838, 18.5h:2.840, 19h:2.830, 19.5h:2.855, 20h:2.844, 20.5h:2.846, 21h:2.840, 21.5h:2.844, 22h:2.829, 22.5h:2.830, 23h:2.825, 23.5h:2.837<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> 0.5%<h2 style=";text-align:left;direction:ltr"> 0h:0, 0.5h:0.133, 1h:0.186, 1.5h:0.328, 2h:0.581, 2.5h:0.845, 3h:1.2 12, 3.5h:1.651, 4h:1.895, 4.5h:2.070, 5h:2.193, 5.5h:2.269, 6h:2.319, 6.5h:2.347, 7h:2.371, 7.5h:2.374, 8h:2.382, 8.5h:2.368, 9h:2.336, 9.5 h:2.335, 10h:2.327, 10.5h:2.323, 11h:2.307, 11.5h:2.305, 12h:2.300, 1 2.5h:2.276, 13h:2.255, 13.5h:2.266, 14h:2.245, 14.5h:2.239, 15h:2.22 0, 15.5h:2.216, 16h:2.215, 16.5h:2.217, 17h:2.204, 17.5h:2.199, 18h:2 .195, 18.5h:2.193, 19h:2.195, 19.5h:2.189, 20h:2.184, 20.5h:2.204, 21 h:2.194, 21.5h:2.197, 22h:2.203, 22.5h:2.198, 23h:2.210, 23.5h:2.193 As shown in Figure 8, it was found that the OD660 value decreased depending on the concentration of PE contained in the LB liquid medium. These results demonstrate that PE exhibits a growth inhibitory effect on Staphylococcus aureus.

[0060] (Test Example 2) Growth Inhibition Test 2 Staphylococcus aureus was cultured in the same manner as in Test Example 1 of Example 3. After 23.5 hours from the start of the culture, the absorbance at 660 nm (OD660) was measured. After the measurement, the blank OD660 value was subtracted from the OD660 value at each concentration to calculate the value (n=3). These results are shown in Figures 9 and 10.

[0061] Fig. 9 is a graph showing the results of a test of the growth inhibition of JCM 2151 strain by various concentrations of PE. In Fig. 9, the vertical axis shows the OD660 value, and the horizontal axis shows the polyol ester concentration (%). The values ​​on the vertical axis in Figure 9 are as follows: OD660 value 0%: 2.932, 0.03125%: 2.872, 0.0625%: 2.603, 0.125%: 2.001, 0.25%: 1.465, 0.5%: 0.698 Error range 0%: 0.179, 0.03125%: 0.179, 0.0625%: 0.038, 0.125%: 0.055, 0.25%: 0.254, 0.5%: 0.142 As shown in FIG. 9, it was found that the OD660 value decreased depending on the concentration of PE contained in the LB liquid medium.

[0062] Fig. 10 is a graph showing the results of a test of the effect of various concentrations of PE on the growth inhibition of strain JCM 2874. In Fig. 10, the vertical axis shows the OD660 value, and the horizontal axis shows the polyol ester concentration (%). The values ​​on the vertical axis in Figure 10 are as follows: OD660 value 0%: 2.870, 0.03125%: 3.030, 0.0625%: 2.770, 0.125%: 2.319, 0.25%: 1.768, 0.5%: 0.969 Error range 0%: 0.179, 0.03125%: 0.179, 0.0625%: 0.038, 0.125%: 0.055, 0.25%: 0.254, 0.5%: 0.142 10, it was found that the OD660 value decreased depending on the concentration of PE contained in the LB liquid medium. These results demonstrate that PE exhibits a growth inhibitory effect on Staphylococcus aureus.

[0063] (Test Example 3) Colony count measurement The number of colonies formed was measured using the Staphylococcus aureus culture solution after the growth inhibition test. Specifically, the Staphylococcus aureus culture solution after the growth inhibition test in Test Example 1 of Example 2 was collected, and a series of dilutions was prepared every 10 times using LB liquid medium. After the preparation, 100 μl of each was dropped onto an LB agar medium containing 1.5% agar, and the whole surface was spread using a cone-larger. After the spreading, the plates were cultured under conditions of 37° C. and 16 hours using a thermostatic incubator. After the culture, the plates of each dilution series were observed and the number of colonies (colony forming unit: CFU / ml) was measured. These results are shown in Tables 2 and 3 below.

[0064] [Table 2]

[0065] The above Table 2 shows the results of the colony count for the JCM 2151 strain. As shown in the above Table 2, it was found that the number of colonies formed decreased depending on the concentration of PE contained in the LB liquid medium.

[0066] [Table 3]

[0067] Table 3 shows the results of the colony count for the JCM 2874 strain. As shown in Table 3, it was found that the number of colonies formed decreased depending on the concentration of PE contained in the LB liquid medium. These results demonstrated that PE has antibacterial activity against Staphylococcus aureus.

[0068] This application claims priority based on Japanese Patent Application No. 2023-101864, filed on June 21, 2023, the disclosure of which is incorporated herein in its entirety. [Industrial Applicability]

[0069] The present invention can be used, for example, in compositions (cosmetics and the like) for removing or inhibiting the formation of biofilms formed by Staphylococcus aureus and / or Propionibacterium acnes on the skin of humans and the like.

Claims

1. The following formula (I): 【Chemistry 1】 (In the formula, X is an alditol residue in which one primary hydroxyl group has been removed from an alditol and one or more hydroxyl groups may be acetylated, and the alditol is mannitol or arabitol; R ２ is C １３ H ２７ Or C １５ H ３１ It is a straight chain hydrocarbon group represented by the formula: A cosmetic composition for removing or inhibiting the formation of a biofilm formed by Staphylococcus aureus and / or Propionibacterium acnes, comprising a polyol ester represented by the formula:

2. The cosmetic composition according to claim 1 for imparting antibacterial activity against Staphylococcus aureus and / or Propionibacterium acnes.

3. The cosmetic composition according to claim 1 or 2, comprising at least 0.1% by mass of the polyol ester.

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