Method for manufacturing polar lipid fermentation product using skin flora and cosmetic composition comprising same

Abstract

The present invention relates to a method for providing a lipid fermentation product having high polarity using skin flora. The lipid fermentation product of the present invention contains glycolipids with high polarity, and thus can easily form emulsified self-assembled particles and liquid crystal emulsions to improve emulsification properties.

Description

Method for producing polar lipid fermentation product using skin-resident bacteria and cosmetic composition containing the same

[0001] This application claims the benefit of priority to Korean Patent Application No. 10-2024-0085980, filed July 1, 2024, the entire contents of which are incorporated herein by reference.

[0002] The present invention relates to a method for producing a highly polar lipid fermentation product using a specific strain containing a lipid-decomposing enzyme. The present invention also relates to a cosmetic composition comprising the fermentation product.

[0003] The skin is composed of the epidermis, dermis, and subcutaneous fat layer. The epidermis, the outermost layer, acts as a protective barrier and is responsible for the skin's immune function. Skin is inhabited by commensal microorganisms, primarily found in the outermost layer of the epidermis and the upper portion of hair follicles (supraderial layer). Approximately 1,000 species of microorganisms exist within the skin commensal flora, and aerobic microorganisms can secrete lipolytic enzymes that utilize epidermal lipids.

[0004] A variety of substances are used to moisturize the skin and deliver active ingredients. To apply these substances as cosmetics, surfactants are typically used to solubilize the two-phase materials, which are difficult to mix. Synthetic surfactants offer excellent solubilizing properties, but they can be irritating to the skin. Natural surfactants, while skin-friendly, can have lower solubilizing properties and emulsification stability than synthetic surfactants.

[0005] Therefore, research on natural surfactants is necessary to achieve stabilization of insoluble substances, improved dispersibility, and high skin absorption rate.

[0006] The purpose of the present invention is to provide a cosmetic composition that improves skin affinity and skin absorption rate of effective substances by producing a polar lipid fermentation product using skin-resident bacteria.

[0007] In order to solve the above problem, the present invention,

[0008] Step of culturing a microorganism with the deposit number KCTC 15741BP (Korea Microbial Resource Center) as a skin flora;

[0009] Pre-culture stage for culturing the spawn culture;

[0010] The first main culture stage of culturing the pre-culture; and

[0011] Includes a second main culture step of fermenting the first main culture,

[0012] A method for producing a polar lipid fermentation product is provided, wherein the fermentation product contains glycolipids.

[0013] According to one embodiment, the starter culture medium may include casein, soybean, dextrose, sodium chloride, and dibasic potassium phosphate. Specifically, the starter culture medium may include 1 to 40 g / L of casein, 0.5 to 30 g / L of soybean, 0.1 to 20 g / L of dextrose, 0.5 to 40 g / L of sodium chloride, and 0.1 to 20 g / L of dibasic potassium phosphate.

[0014] According to one embodiment, the pre-culture medium may include soybeans, yeast extract, glycerin, monobasic potassium phosphate, dibasic potassium phosphate, and vegetable oil. Specifically, the pre-culture medium may include 0.01 to 10 g / L of soybeans, 1 to 20 g / L of yeast extract, 0.5 to 40 g / L of glycerin, 0.5 to 40 g / L of monobasic potassium phosphate, 0.3 to 30 g / L of dibasic potassium phosphate, and 0.1 to 10 g / L of vegetable oil.

[0015] According to one embodiment, the primary main culture medium may include soybeans, yeast extract, glycerin, monobasic potassium phosphate, dibasic potassium phosphate, and vegetable oil. Specifically, the primary main culture medium may include 0.01 to 10 g / L of soybeans, 10 to 40 g / L of yeast extract, 0.5 to 40 g / L of glycerin, 0.5 to 40 g / L of monobasic potassium phosphate, 0.3 to 30 g / L of dibasic potassium phosphate, and 10 to 50 g / L of vegetable oil.

[0016] According to one embodiment, the seed culturing step, pre-cultivation step, primary main culturing step or secondary main culturing step may include a step of culturing under aerobic conditions of 50 to 300 rpm and 10 to 100 NL / min at 23 to 40°C for 10 to 40 hours.

[0017] According to one embodiment, the first main culture step comprises a total viable cell count of 1.0 X 10 10 ~ 6.0 X 10 10 The second main culture step may include a step of culturing by adding sugar alcohol, monobasic potassium phosphate, dibasic potassium phosphate, and vegetable oil to the first main culture. Specifically, the second main culture step may include a step of culturing by adding 10 to 300 g / L of sugar alcohol, 0.5 to 40 g / L of monobasic potassium phosphate, 0.5 to 40 g / L of dibasic potassium phosphate, and 50 to 500 g / L of vegetable oil.

[0018] According to one embodiment, the sugar alcohol may include one or more of saccharide, dextrose, xylitol, mannitol, sorbitol, trehalose, erythritol, sucrose, fructose, maltose, and lactose.

[0019] According to another embodiment of the present invention, a polar lipid fermentation product manufactured by the method described above is provided.

[0020] According to another embodiment of the present invention, a cosmetic composition comprising the polar lipid fermentation product is provided.

[0021] Specific details of other implementation examples according to the present invention are included in the detailed description below.

[0022] The polar lipid fermentation product according to the present invention contains highly polar glycolipids through cultivation using skin-resident bacteria. Furthermore, the inclusion of highly polar glycolipids facilitates the formation of emulsifying self-assembled particles and liquid crystal emulsions, thereby enhancing emulsifying properties.

[0023] Figure 1 is a graph showing the appropriate time for substrate introduction for secondary culture.

[0024] Figure 2 is a photograph showing a fermented product according to Example 1.

[0025] Figures 3 and 4 are photographs showing the TLC comparison results of Comparative Example 1 and Example 1.

[0026] Figure 5 is a graph showing the results of analyzing the particle size of the fermented product of Example 1.

[0027] Figure 6 is a photograph showing the shape of the fermented product particles of Example 1.

[0028] Figure 7 is a photograph showing the liquid crystal emulsification ability of the fermented product of Example 1.

[0029] Figure 8 is a photograph confirming the solubility of the fermented product of Example 1.

[0030] The present invention is susceptible to various modifications and embodiments, and thus specific embodiments will be illustrated and described in detail. However, this is not intended to limit the present invention to specific embodiments, but rather to encompass all modifications, equivalents, and alternatives falling within the spirit and technical scope of the present invention. In describing the present invention, detailed descriptions of related known technologies will be omitted if they are deemed to obscure the gist of the present invention.

[0031] Hereinafter, a method for producing a polar lipid fermentation product using skin-resident bacteria according to the present invention and a cosmetic composition comprising the same will be described. The present invention utilizes skin-resident bacteria, which are among the various microorganisms that inhabit the skin and have a beneficial effect on the skin, and coexist with it. The composition of the medium was optimized to produce highly polar glycolipids from skin-resident bacteria.

[0032] Specifically, the present invention provides a method for producing a polar lipid fermentation product, comprising: a step of culturing a skin-resident fungus; a pre-culture step of culturing the pre-culture; a first main culture step of culturing the pre-culture; and a second main culture step of fermenting the first main culture.

[0033] The skin-resident bacteria of the present invention may be a strain of Epidermidibacterium keratini sp., and the microorganism with the deposit number KCTC 15741BP (Korea Center for Microbiological Resources) may be used.

[0034] According to one embodiment, the starter culture medium may include casein (Pancreatic Digest of Casein), soybean (Papain Digest of Soybean), dextrose, sodium chloride, and dipotassium phosphate. Specifically, the starter culture medium may comprise 1 to 40 g / L, for example, 10 to 30 g / L, or 10 to 20 g / L, casein, 0.5 to 30 g / L, for example, 1 to 20 g / L, 1 to 10 g / L, or 1 to 5 g / L, soybean, 0.1 to 20 g / L, for example, 1 to 10 g / L, or 1 to 5 g / L, dextrose, 0.5 to 40 g / L, for example, 1 to 30 g / L, 1 to 20 g / L, or 1 to 10 g / L, sodium chloride, and 0.1 to 20 g / L, for example, 1 to 10 g / L, or 1 to 5 g / L, potassium phosphate dibasic.

[0035] According to one embodiment, the pre-culture medium may include papain digest soybean, yeast extract, glycerin, monopotassium phosphate, dipotassium phosphate, and vegetable oil. Specifically, for example, the pre-culture medium may comprise soybean 0.01 to 10 g / L, for example 0.01 to 5 g / L, 0.01 to 1 or 0.05 to 3 g / L, yeast extract 1 to 20 g / L, for example 1 to 15 or 5 to 15 g / L, glycerin 0.5 to 40 g / L, for example 1 to 30 g / L, 1 to 20 g / L or 1 to 10 g / L, potassium phosphate monobasic 0.5 to 40 g / L, for example 1 to 30 g / L, 1 to 20 g / L, 1 to 10 g / L, potassium phosphate dibasic 0.3 to 30 g / L, for example 1 to 20 g / L, 1 to 10 g / L or 1 to 5 g / L and vegetable oil 0.1 to 10 g / L, for example, 0.1 to 5 g / L or 0.1 to 3 g / L.

[0036] According to one embodiment, the primary culture medium may include soybean (Papain digest soybean), yeast extract, glycerin, monopotassium phosphate, dipotassium phosphate, and vegetable oil. Specifically, for example, the primary main culture medium may comprise soybean 0.01 to 10 g / L, for example 0.01 to 5 g / L or 0.01 to 1 g / L, yeast extract 10 to 40 g / L, for example 10 to 30 g / L or 15 to 25 g / L, glycerin 0.5 to 40 g / L, for example 1 to 30 g / L, 1 to 20 g / L or 1 to 10 g / L, potassium phosphate monobasic 0.5 to 40 g / L, for example 0.5 to 30 g / L, 0.5 to 20 g / L or 1 to 10 g / L, potassium phosphate dibasic 0.3 to 30 g / L, for example 1 to 20 g / L or 1 to 10 g / L and vegetable oil 10 to 50 g / L, for example, 10 to 40 g / L, 10 to 30 g / L or 15 to 25 g / L.

[0037] According to one embodiment, the seed culturing step, the pre-culturing step, the first main culture step or the second main culture step may include a step of culturing under aerobic conditions at 23 to 40°C, for example, 25 to 35°C, or 28 to 34°C, at 50 to 300 rpm, for example, or 100 to 200 rpm, at 10 to 100 NL / min, for example, 10 to 80 NL / min, 10 to 50 NL / min or 20 to 40 NL / min, for 10 to 40 hours, for example, 10 to 30 hours, 15 to 28 hours, or 20 to 25 hours. The total culturing time of the first main culture step and the second main culture step may not exceed 100 hours.

[0038] According to one embodiment, the first main culture step comprises a total viable cell count of 1.0 X 10 10 ~ 6.0 X 10 10 It can be terminated at the point where CFU / ml is reached. In addition, the secondary main culture step may include a step of culturing by adding sugar alcohol, potassium phosphate monobasic, potassium phosphate dibasic, and vegetable oil to the primary main culture. Specifically, the secondary main culture step may include a step of culturing by adding 10 to 300 g / L of sugar alcohol, for example 50 to 200 g / L or 50 to 150 g / L, 0.5 to 40 g / L of potassium phosphate monobasic, for example 1 to 30 g / L, 1 to 20 g / L or 1 to 10 g / L, 0.5 to 40 g / L of potassium phosphate dibasic, for example 1 to 30 g / L, 1 to 20 g / L or 1 to 10 g / L and 50 to 500 g / L of vegetable oil, for example 50 to 400 g / L, 100 to 300 g / L or 150 to 250 g / L.

[0039] According to one embodiment, the sugar alcohol may include one or more of saccharide, dextrose, xylitol, mannitol, sorbitol, trehalose, erythritol, sucrose, fructose, maltose, and lactose.

[0040] According to one embodiment, the vegetable oil used in the present invention is not particularly limited as long as it is a vegetable oil without cytotoxicity. Specifically, for example, the vegetable oil may include one or more of edible or human-friendly oils such as macadamia oil, sunflower seed, grape seed, canola, rice germ, olive, soybean, argan, brown rice, perilla, sesame, almond, peanut, corn, red ginseng, avocado, macadamia, coconut, rosehip, vitamin tree seed, shea tree fruit, oil palm, bergamot fruit, camellia seed, safflower seed, apricot kernel, poppy seed, evening primrose seed, castor seed, green tea seed, meadowfoam seed, flax seed, and hemp seed, but is not particularly limited to the above types.

[0041] According to one embodiment, the method for producing a polar lipid fermentation product may further include a separation and purification step after the secondary main culture step. Specifically, the separation step may include a step of allowing the secondary main culture to stand at 50 to 80°C, for example, 65 to 75°C, to separate an oil layer and a water layer. In addition, the purification step may include a step of recovering the separated oil layer and removing any remaining sugars (substrates), moisture, and fatty acids.

[0042] The polar lipid fermentation product manufactured by the above method has a high content of highly polar glycolipids.

[0043] According to another embodiment of the present invention, a cosmetic composition comprising a polar lipid fermentation product as described above is provided.

[0044] According to one embodiment, the cosmetic composition of the present invention may include ingredients commonly used in cosmetic compositions, such as stabilizers, solubilizers, vitamins, pigments, fragrances, adjuvants, and carriers.

[0045] In addition, it can be manufactured in any formulation that is commonly manufactured in the art and can be applied dermatologically. Being applicable dermatologically means that it is a composition that can have an effective effect that is relatively non-toxic and harmless to the target of application, and may include an external agent that can be applied to the skin, and it can mean that the side effects resulting from the composition do not reduce the efficacy of the active ingredient, do not cause serious irritation to the target of application, and do not damage the activity and physical properties of the active ingredients. The cosmetic composition applicable dermatologically of the present invention can be formulated as, for example, a solution, a suspension, a milky lotion, an emulsion, a paste, a gel, a pack, a cream, a lotion, a powder, a soap, a surfactant-containing cleanser, an oil, a powder foundation, an emulsion foundation, a wax foundation, a spray, and a hair cosmetic, but is not limited thereto. Specifically, for example, it can be manufactured in the form of a skin lotion, skin softener, skin toner, astringent, lotion, gel, milk lotion, moisture lotion, nutrition lotion, massage cream, nutrition cream, moisture cream, hand cream, foundation, essence, ampoule, nutrition essence, pack, soap, hair shampoo, foot shampoo, cleansing foam, cleansing lotion, cleansing cream, body lotion, and body cleanser.

[0046] Hereinafter, embodiments of the present invention will be described in detail so that those skilled in the art can easily implement them. However, the present invention may be implemented in various different forms and is not limited to the embodiments described herein.

[0047]

[0048] Manufacturing example: Manufacturing of polar lipid fermentation product

[0049] spawn culture

[0050] A medium containing 17.0 g / L of casein (Pancreatic Digest of Casein), 3.0 g / L of soybean (Papain Digest of Soybean), 2.5 g / L of dextrose, 5.0 g of sodium chloride, 2.5 g / L of dipotassium phosphate, and 970 g / L of purified water was prepared. The prepared medium was inoculated with a microorganism with the deposit number KCTC 15741BP (Korea Center for Microbiological Resources) as a skin flora, and cultured for 24 hours under aerobic conditions at 33°C and 150 rpm to prepare a starter culture.

[0051]

[0052] Pre-culture

[0053] A medium containing 0.1 g / L of soybean (Papain digest soybean), 10.0 g / L of yeast extract, 5.0 g / L of glycerin, 4.5 g of monopotassium phosphate, 3.0 g / L of dipotassium phosphate, 1 g / L of macadamia seed oil, and 976.4 g / L of purified water was prepared.

[0054] A pre-culture was prepared by adding 10% of the inoculum and culturing it for 24 hours at a temperature of 33°C and a stirring speed of 180 rpm under aerobic conditions of 30 NL / min.

[0055]

[0056] Primary culture

[0057] A medium containing 0.1 g / L of soybean (Papain digest soybean), 20.0 g / L of yeast extract, 5.0 g / L of glycerin, 4.5 g of monopotassium phosphate, 3.0 g / L of dipotassium phosphate, 20.0 g / L of vegetable oil, and 947.4 g / L of purified water was prepared. 10% of the above spawn culture was added, and the primary main culture was prepared by culturing for 24 hours at a culture temperature of 33°C and a stirring speed of 100 rpm under aerobic conditions of 30 NL / min. The primary main culture was cultured until it reached the logarithmic growth phase, and the primary main culture was terminated when the total viable cell count was 1.0 X 10 10 ~ 6.0 X 10 10 This is the point where CFU / ml is reached.

[0058]

[0059] Secondary main culture

[0060] Secondary main culture was conducted using a fed-batch method by mixing and sterilizing 100 g / L sugar alcohol and 200 g / L vegetable oil into the primary main culture. However, the sugar alcohol was dissolved in potassium phosphate buffer and added. The specific composition of the secondary main culture medium is as shown in Table 1 or Table 2.

[0061] The end point of the secondary main culture is the point of consumption of the oil substrate. The secondary main culture was performed until the oil substrate was completely consumed by collecting a sample of the secondary main culture during cultivation and centrifuging it. The primary and secondary main cultures were cultured for a total of up to 100 hours. The secondary main culture was cultured when the microbial activity of the primary main culture was the highest, by adding the secondary main culture medium to increase the efficiency of polar lipid fermentation product production. The optimal time for substrate addition from the primary to the secondary main culture was 18 to 26 hours after the start of the primary culture, and the optimal time for substrate addition is shown graphically in Figure 1.

[0062]

[0063] Separation and purification

[0064] The secondary main culture was centrifuged at high speed to remove microbial cells, and 1N HCl solution was added to the secondary main culture from which the cells had been removed to separate the aqueous phase and the oil phase. The separated aqueous phase was discarded, and the oil phase was recovered, and 95% ethanol corresponding to twice its weight was added, and the mixture was stirred at 200 rpm for 4 hours. The mixture was allowed to settle, and the ethanol layer solution was recovered and concentrated to obtain a highly polar lipid fermentation product. The state of the fermentation product according to Example 1 is shown in Fig. 2.

[0065]

[0066] Production of fermented products according to the type of sugar alcohol substrate

[0067] The medium used for the second main culture was applied according to the composition in Table 1.

[0068] Step Medium Composition (g / L) Example 1 Example 2-1 Example 2-2 Example 2-3 Example 2-4 Example 2-5 Example 2-6 Example 2-7 Example 2-8 Example 2-9 Example 2-10 Secondary Culture Saccharide isomerate 100.0----------Dextrose-100.0---------Xylitol--100.0--------Mannitol---100.0-------Sorbitol----100.0-------Trehalose-----100.0-----Erythritol------100.0----Sucrose-------100.0---Fructose--------100.0--Maltose--------100.0-Lactose--------100.0Monopotassium phosphate4.54.54.54.54.54.54.54.54.54.54.5Dipotassium Phosphate3.03.03.03.03.03.03.03.03.03.03.0Macadamia seed Oil200.0200.0200.0200.0200.0200.0200.0200.0200.0200.0200.0Water692.5692.5692.5692.5692.5692.5692.5692.5692.5692.5692.5

[0069]

[0070] Production of fermented products according to oil type

[0071] The medium used for the second main culture was applied according to the composition in Table 2.

[0072] Step Medium Composition (g / L) Example 1 Example 3-1 Example 3-2 Example 3-3 Example 3-4 Example 3-5 Example 3-6 Example 3-7 Example 3-8 Example 3-9 Example 3-10 Secondary Culture Saccharide isomerate 100.0100.0100.0100.0100.0100.0100.0100.0100.0100.0100.0100.0100.0100.0100.0Monopotassium phosphate 4.54.54.54.54.54.54.54.54.54.54.54.54.54.54.5 Dipotassium Phosphate 3.03.03.03.03.03.03.03.03.03.03.0Macadamia seed Oil 200.0Olive Oil200.0Sunflower seed Oil200.0Grape seed oil200.0Rice germ Oil200.0Coconut Oil200.0Apricot kernel Oil200.0Camellia Japonica seed Oil200.0Argan Oil200.0Safflower Oil200.0Avocado Oil200.0Water692.5692.5692.5692.5692.5692.5692.5692.5692.5692.5692.5

[0073]

[0074] Comparative Example 1

[0075] spawn culture

[0076] A microorganism with the deposit number KCCM 11843P (Korea Center for Microorganism Conservation (Overseas), 2016.06.08) was cultured as a skin flora. The culture was prepared by inoculating R2A (BD Difco, USA) medium and shaking culture under aerobic conditions at 25℃. Specifically, the medium composition was casein acid hydrolysate 0.5 g / L, yeast extract 0.5 g / L, glucose 0.5 g / L, soluble starch 0.5 g / L, K2HPO4 0.3 g / L, sodium pyruvate 0.3 g / L, casein peptone 0.5 g / L, and MgSO4 0.05 g / L.

[0077] Pre-culture

[0078] 100 g / L of the spawn culture solution was added to the same medium as the spawn culture medium composition above, and cultured under aerobic conditions of 20°C, 15 NL / min, and 50 rpm to prepare a pre-culture. In addition, the fermentation time point was set for the pre-culture by measuring turbidity. Specifically, the pre-culture was sampled, diluted 5% with 0.85% NaCl solution, and then the absorbance was measured at a wavelength of 600 nm using a spectrophotometer (Spectrophotometer, BioTek, USA). The end point of the pre-culture and the inoculation point of the main fermentation were set based on the absorbance range of 0.2 to 0.4.

[0079] Main fermentation

[0080] Glycerol 7.5 g / L, yeast extract 5 g / L, casein peptone 1 g / L, K2HPO4 0.45 g / L, KH2PO4 0.3 g / L, MgSO 4·The temperature of the main fermentation medium containing 7H2O 0.02 g / L, NaCl 1 g / L, NH2SO4 0.5 g / L, KNO 0.1 g / L, FeSO4 0.01 g / L, macadamia oil 500 g / L, and purified water 484.12 g / L was set to 20°C, and 10 wt% of the above pre-culture was inoculated, and fermentation was performed under aerobic conditions of 20°C, 300 NL / min, and 600 rpm to produce a fermented product (fermented oil).

[0081] Separation and purification

[0082] To obtain the fermented oil from the above fermented product, the fermented product was centrifuged to separate the oil layer. The oil layer was recovered, and 100 g / L of MgSO4 was added to remove any remaining moisture and impurities, followed by stirring. After stirring for 2 hours, the mixture was filtered using a filter pad (CH-ST-150, Hyundai Micro, Korea) in a filter press. The filtered oil was finally subjected to a sterile filtration (0.2 μm filter) to obtain the fermented oil.

[0083]

[0084] Experimental Example 1: TLC Lipid Analysis

[0085] The first condition (Condition 1) was that 2 μl of the sample was diluted 20% in chloroform on a silica gel TLC plate (TLC Silica gel 60, Merck; 5 cm x 6.5 cm) and developed with a mobile phase solvent containing chloroform, methanol, and acetone (12:0.5:0.1, v / v) until the end point. The naturally dried plate was sprayed with 10% sulfuric acid mixed with ethanol, dried, and heated at 100°C for 5 minutes to visualize the lipid spot. The TLC comparison results of Comparative Example 1 and Example 1 according to Condition 1 are shown in Figure 3.

[0086] The second condition (Condition 2) was prepared by preparing a silica gel TLC plate and sample in the same manner, and then developed until the end point with a mobile phase solvent containing chloroform, methanol, and water (6.5:3.5:0.4, v / v). Orcinol solution (0.2% Orcinol solution) was sprayed onto the naturally dried plate, dried, and heated at 100°C for 5 minutes to visualize lipid spots. Orcinol solution is a glycolipid chromogenic reagent, and non-glycolipids do not develop color. The TLC comparison results of Comparative Example 1 and Example 1 according to Condition 2 are shown in Figure 4.

[0087] As shown in Figures 3 and 4, it was confirmed that the glycolipid spot (component) was colored in Example 1 compared to Comparative Example 1.

[0088]

[0089] Experimental Example 2: Evaluation of Polyhydric Alcohol Miscibility

[0090] The miscibility of the polar lipid fermentation product according to Example 1 with polyhydric alcohol, a cosmetic solvent, was confirmed, and the results are shown in Table 3.

[0091] Polyhydric alcohol concentration (%) Example 1 Comparative example 1 Glycerin 400 1,3 Propanediol 300 Dipropylene glycol 50 5 Less than 1,3 Butylene glycol 400 Pentylene glycol 50 5 Less than 1,2 Hexanediol 50 5 Less than 2,3 Butylene glycol 300

[0092] As shown in Table 3, Example 1 is composed of lipids with higher polarity than Comparative Example 1, and thus has a higher solubility in polyalcohol.

[0093]

[0094] Experimental Example 3: Confirmation of Self-Assembly Particle Formation

[0095] The polar lipid fermentation product according to Example 1 and pentylene glycol were mixed and dissolved in a 1:1 ratio and added to purified water at 1% (v / v). Then, the shape and size of the particles of the composition were measured.

[0096] Particle size analysis was performed using a Zetasiser Lab. (Malvern Panalytical) model instrument. The results are shown in Fig. 5, and the average particle size was confirmed to be 43 nm.

[0097] In addition, the shape of the self-assembled particles was observed using a transmission electron microscope (Jem-f-200, JEOL). The sample composition was dropped onto a carbon-coated copper grid (TF300C, TMA) and pretreated by drying at room temperature for 24 hours. The observation image is shown in Fig. 6, and it was confirmed that spherical emulsified particles with a size of approximately 50 nm were formed.

[0098]

[0099] Experimental Example 4: Confirmation of Liquid Crystal Emulsification Ability

[0100] To confirm the liquid crystal emulsifying ability of the polar lipid fermentation product according to Example 1, a composition was prepared with the composition shown in Table 4. Specifically, Phases A and B were completely dissolved by heating them to 80°C in a water bath. Then, Phase A was added to Phase B, and emulsified using a homogenizer at 3000 rpm for 3 minutes. The completed composition was cooled to 30°C. The performance was compared with that of Comparative Example 1.

[0101] Phase Component Content (%)AExample 15.0-Comparative Example 1-5.0Pentylene glycol5.05.0Macadamia seed Oil10.010.0Oleyl alcohol0.80.8Stearyl behenate5.05.0BGlycerin5.05.01,2-Hexanediol2.02.0Water67.267.2

[0102] The results were observed at 400x magnification using a polarizing microscope (BX53, Olympus, Japan) and are shown in Fig. 7. As shown in Fig. 7, Example 1 had a constant liquid crystal formation and size, while Comparative Example 1 did not produce a liquid crystal structure (Maltese cross).

[0103]

[0104] Experimental Example 5: Solubility Verification

[0105] In order to confirm the solubility of the polar lipid fermentation product according to Example 1, a toner was prepared using a fragrance that does not readily dissolve in water.

[0106] According to the composition in Table 5, Phase B was prepared by heating to 50°C, and Phase A was prepared by sufficient mixing and stirring at 40°C. The prepared Phase A was added to Phase B while stirring, and mixed for 5 minutes after addition. After cooling, the solubilization ability was confirmed. The results are shown in Fig. 8. The composition according to Comparative Example 1 was confirmed to have oil separation and turbidity, whereas the composition according to Example 1 was confirmed to have no oil separation and to be transparent.

[0107] Phase Ingredient Content (%) A Example 10.15 - Comparative Example 1-015 Pentylene glycol 0.20 0.20 1,2-Hexanediol 0.10 0.10 Propanediol 0.05 0.05 Fragrances 0.05 0.05 B 1,3 Butylene glycol 5.00 5.00 1,2-Hexanediol 2.00 2.00 Glycerin 10.00 10.00 Water 82.45 82.45

[0108]

[0109] As described above, the present invention can produce a highly polar lipid fermentation product. The polar lipid fermentation product of the present invention comprises at least 11 types of polar lipids, exhibits excellent solubility in polyhydric alcohols, and readily forms emulsifying self-assembled particles in aqueous solutions. Therefore, stabilization of oil-soluble and poorly soluble substances, improved dispersibility, and high skin absorption can be expected. Furthermore, it can be applied to fatty alcohols and waxes to easily produce liquid crystal emulsions, and can solubilize effective substances such as fragrances and essential oils without using ethanol.

[0110] As described above, specific parts of the present invention have been described in detail. For those skilled in the art, these specific descriptions are merely preferred embodiments, and the scope of the present invention is not limited to the specific embodiments described above.

[0111]

[0112]

[0113] [Microorganism Deposit Certificate]

[0114]

Claims

1. Step of culturing the microorganism with the deposit number KCTC 15741BP (Korea Microbial Resource Center) as a skin flora; Pre-culture stage for culturing the spawn culture; The first main culture stage of culturing the pre-culture; and Includes a second main culture step of fermenting the first main culture, A method for producing a polar lipid fermentation product, wherein the fermentation product contains glycolipids.

2. In paragraph 1, A method for producing a polar lipid fermentation product, wherein the starter culture medium contains casein, soybeans, dextrose, sodium chloride and potassium phosphate dibasic.

3. In paragraph 1, A method for producing a polar lipid fermentation product, wherein the starter culture medium comprises 1 to 40 g / L of casein, 0.5 to 30 g / L of soybean, 0.1 to 20 g / L of dextrose, 0.5 to 40 g / L of sodium chloride, and 0.1 to 20 g / L of potassium phosphate dibasic.

4. In paragraph 1, A method for producing a polar lipid fermentation product, wherein the pre-culture medium comprises soybeans, yeast extract, glycerin, potassium phosphate monobasic, potassium phosphate dibasic and vegetable oil.

5. In paragraph 1, A method for producing a polar lipid fermentation product, wherein the pre-culture medium comprises 0.01 to 10 g / L of soybean, 1 to 20 g / L of yeast extract, 0.5 to 40 g / L of glycerin, 0.5 to 40 g / L of potassium phosphate monobasic, 0.3 to 30 g / L of potassium phosphate dibasic, and 0.1 to 10 g / L of vegetable oil.

6. In paragraph 1, A method for producing a polar lipid fermentation product, wherein the primary culture medium comprises soybeans, yeast extract, glycerin, potassium phosphate monobasic, potassium phosphate dibasic, and vegetable oil.

7. In paragraph 1, A method for producing a polar lipid fermentation product, wherein the primary culture medium comprises 0.01 to 10 g / L of soybean, 10 to 40 g / L of yeast extract, 0.5 to 40 g / L of glycerin, 0.5 to 40 g / L of potassium phosphate monobasic, 0.3 to 30 g / L of potassium phosphate dibasic, and 10 to 50 g / L of vegetable oil.

8. In paragraph 1, A method for producing a polar lipid fermentation product, wherein the above-mentioned seed culturing step, pre-cultivation step, first main culture step or second main culture step includes a step of culturing at 23 to 40°C, 50 to 300 rpm, and 10 to 100 NL / min under aerobic conditions for 10 to 40 hours.

9. In paragraph 1, The above first main culture step has a total viable cell count of 1.0 X 10 10 ~ 6.0 X 10 10 It ends at the point where CFU / ml is reached, A method for producing a polar lipid fermentation product, wherein the second main culture step includes a step of culturing by adding sugar alcohol, potassium phosphate monobasic, potassium phosphate dibasic and vegetable oil to the first main culture.

10. In paragraph 9, A method for producing a polar lipid fermentation product, wherein the second main culture step comprises a step of culturing by adding 10 to 300 g / L of sugar alcohol, 0.5 to 40 g / L of potassium phosphate monobasic, 0.5 to 40 g / L of potassium phosphate dibasic, and 50 to 500 g / L of vegetable oil.

11. In paragraph 9, A method for producing a polar lipid fermentation product, wherein the sugar alcohol comprises at least one of saccharide, dextrose, xylitol, mannitol, sorbitol, trehalose, erythritol, sucrose, fructose, maltose, and lactose.

12. A polar lipid fermentation product manufactured by a method according to any one of claims 1 to 11.

13. A cosmetic composition comprising the polar lipid fermentation product of Article 12.

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