Application of phenyllactic acid to improvement of skin moisture state

Abstract

The invention discloses a composition for moisturizing skin or improving skin moisture state and a preparation method of phenyllactic acid. A composition for moisturizing or improving the moisture state of skin according to an embodiment of the present invention comprises: a composition selected from phenyllactic acid; a strain having the phenyllactic acid producing ability; lysates thereof; a culture thereof; fermentation products thereof; and at least one of the bacterial strain, lysate, culture or fermentation product extract as an active ingredient, the composition can effectively increase the expression of hyaluronic acid synthase in skin cells, thereby protecting the skin from a dry state caused by blue light, ultraviolet light and the like. Therefore, the composition can be applied to a cosmetic composition, a medicine composition, a general food composition or a health food composition for moisturizing.

Description

Technical Field

[0001] The invention discloses a composition for improving the moisture state of skin and a preparation method of phenyllactic acid. Background Art

[0002] Phenyllactic acid (PLA) is a metabolite derived from phenylalanine. After forming phenylpyruvate as an intermediate metabolite, it is converted to PLA by lactate dehydrogenase. Phenyllactic acid can be found in various fermented foods and lactic acid bacteria cultures. Phenyllactic acid is known to inhibit the growth of harmful bacteria such as Klebsiella pneumonia, Staphylococcus aureus, and Aspergillus flavus. Based on this antimicrobial activity, phenyllactic acid is used as an additive in food and animal feed to achieve long-term preservation.

[0003] Regarding the skin-related effects of phenyllactic acid, a skin external preparation for improving wrinkles is known (US5643953A), and it is known to improve skin tone by inhibiting tyrosinase activity. However, its effects related to moisturizing are still unclear.

[0004] Furthermore, hyaluronic acid, a type of glycosaminoglycan, is a chain-like, high-molecular-weight polysaccharide composed of repeatedly linked glucuronic acid and N-acetylglucosamine residues. Hyaluronic acid is a major component of the extracellular matrix and is reportedly involved in water retention, maintaining intercellular spaces, and storing and diffusing cell growth factors and nutrients. It also participates in cell division, differentiation, and migration. Therefore, increasing hyaluronic acid synthesis may contribute to skin moisturization.

[0005]

Prior art literature

[0006] [Patent Literature]

[0007] (Patent Document 1) U.S. Registered Patent No. 5,643,953A. Summary of the Invention

[0008] Technical issues

[0009] An object of the present invention is to provide a composition for moisturizing the skin or improving the moisture status of the skin.

[0010] Another object of the present invention is to provide a method for preparing phenyllactic acid.

[0011] Technical Solution

[0012] In order to achieve the above-mentioned objectives, in one aspect, the present invention provides a composition for moisturizing the skin, improving the moisture status of the skin, or promoting the production of hyaluronic acid, comprising at least one selected from phenyllactic acid; a strain having the ability to produce the phenyllactic acid; its lysate; its culture; its fermentation product; and an extract of the strain, lysate, culture or fermentation product as an active ingredient.

[0013] In another aspect, the present invention provides a method for preparing phenyllactic acid, comprising the following steps: culturing a Lactobacillus plantarum strain.

[0014] Beneficial effects

[0015] According to one embodiment of the present invention, the composition comprises at least one selected from phenyllactic acid; a strain having the ability to produce phenyllactic acid; a lysate thereof; a culture thereof; a fermentation product thereof; and an extract of the strain, lysate, culture or fermentation product as an active ingredient. The composition can effectively increase the expression of hyaluronic acid synthase in skin cells, thereby protecting the skin from dryness caused by blue light, ultraviolet rays, etc., and can therefore be applied to cosmetic compositions, quasi-pharmaceutical compositions, general food compositions or health food compositions for moisturizing the skin or improving the moisture status of the skin.

[0016] According to one embodiment of the present invention, the composition comprises at least one selected from phenyllactic acid; a strain having the ability to produce phenyllactic acid; a lysate thereof; a culture thereof; a fermentation product thereof; and an extract of the strain, lysate, culture or fermentation product as an effective ingredient. The composition can effectively increase the expression of hyaluronic acid synthase in an organism (eye), and can therefore be applied to a quasi-pharmaceutical composition or a health food composition.

[0017] Furthermore, when the Lactobacillus plantarum strain is cultured in a medium supplemented with phenylalanine and / or calcium carbonate, phenyllactic acid can be efficiently produced. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 Shown are the results of a comparison of the phenyllactic acid-producing abilities of various Lactobacillus strains.

[0019] Figure 2 Shown are the results of a comparison of phenyllactic acid production under different medium compositions and the addition of intermediate metabolites (phenylalanine or phenylpyruvate).

[0020] Figure 3The expression level of the HAS2 (hyaluronan synthase 2) gene under the addition of phenyllactic acid (PLA) or lactobacillus fermentation product is shown. No treatment group: not treated with a stimulus (blue light), - (control group): treated with only a stimulus, PLA: treated with a stimulus and added PLA, Lactobacillus fermentation product: treated with a stimulus and added Lactobacillus fermentation product.

[0021] Figure 4 The figure shows the gene expression level of HAS2 (hyaluronan synthase 2) when a fermentation product of another lactobacillus species, Lactobacillus plantarum APsulloc 331261, was added. Untreated group: no stimulation source (blue light) treatment, - (control group): only stimulation source treatment. DETAILED DESCRIPTION

[0022] Hereinafter, the present invention will be described in more detail.

[0023] In one aspect, the present invention provides a composition for moisturizing the skin, improving the moisture status of the skin, or promoting the production of hyaluronic acid, comprising at least one selected from phenyllactic acid; a strain having the ability to produce the phenyllactic acid; a lysate thereof; a culture thereof; a fermentation product thereof; and an extract of the strain, lysate, culture or fermentation product as an active ingredient.

[0024] In an exemplary embodiment, the strain may be a Lactobacillus plantarum strain.

[0025] In an exemplary embodiment, the strain may be Lactobacillus plantarum APsulloc 331261 (Deposit No.: KCCM11179P).

[0026] In an exemplary embodiment, the strain is capable of converting phenylalanine to phenyllactic acid.

[0027] In an exemplary embodiment, the composition may include phenyllactic acid in an amount of 0.1 ppm or more to 1,000 ppm or less, more specifically, 0.1 ppm or more, 0.2 ppm or more, 0.3 ppm or more, 0.4 ppm or more, 0.5 ppm or more, 0.6 ppm or more, 0.7 ppm or more, 0.8 ppm or more, 0.9 ppm or more, 1 ppm or more, 2 ppm or more, 3 ppm or more, 4 ppm or more, 5 ppm or more, 6 ppm or more, 7 ppm or more, or 1,000 ppm or less, 900 ppm or less, 800 ppm or less. The concentration of hyaluronic acid synthase in skin cells may be less than 10 ppm, less than 700 ppm, less than 600 ppm, less than 500 ppm, less than 400 ppm, less than 300 ppm, less than 200 ppm, less than 100 ppm, less than 90 ppm, less than 80 ppm, less than 70 ppm, less than 60 ppm, less than 50 ppm, less than 40 ppm, less than 30 ppm, less than 20 ppm, less than 19 ppm, less than 17 ppm, less than 16 ppm, less than 15 ppm, less than 14 ppm, less than 13 ppm, less than 12 ppm, less than 11 ppm, or less than 10 ppm, but is not limited thereto. Within the above range, the expression of hyaluronic acid synthase in skin cells can be effectively increased without causing cytotoxicity, skin irritation, etc., thereby protecting the skin from dryness caused by blue light, ultraviolet rays, etc.

[0028] In one exemplary embodiment, the amount of phenyllactic acid administered may be 0.1 to 100 mg / kg / day. If the amount of phenyllactic acid administered is less than 0.1 mg / kg / day, the effect of moisturizing the skin or improving the skin's moisture status, or the effect of promoting hyaluronic acid production may be minimal. If the amount of phenyllactic acid administered is greater than 100 mg / kg / day, cytotoxicity may be exhibited, or the effectiveness of moisturizing the skin or improving the skin's moisture status, or the effect of promoting hyaluronic acid production may be reduced. More specifically, the daily administration amount of phenyllactic acid may be 0.1 mg / kg / day or more, 0.2 mg / kg / day or more, 0.3 mg / kg / day or more, 0.4 mg / kg / day or more, 0.5 mg / kg / day or more, 0.6 mg / kg / day or more, 0.7 mg / kg / day or more, 0.8 mg / kg / day or more, 0.9 mg / kg / day or more, 1.0 mg / kg / day or more, 2.0 mg / kg / day or more, 3.0 mg / kg / day or more, 4.0 mg / kg / day or more, 5.0 mg / kg / day or more, 6.0 mg / kg / day or more, 7.0 mg / kg / day or more, 8.0 mg / kg / day or more, 9.0 mg / kg / day or more, The dosage may be 10.0 mg / kg / day or more, or 100 mg / kg / day or less, 95 mg / kg / day or less, 90 mg / kg / day or less, 85 mg / kg / day or less, 80 mg / kg / day or less, 75 mg / kg / day or less, 70 mg / kg / day or less, 65 mg / kg / day or less, 60 mg / kg / day or less, 55 mg / kg / day or less, 50 mg / kg / day or less, 45 mg / kg / day or less, 40 mg / kg / day or less, 35 mg / kg / day or less, 30 mg / kg / day or less, 25 mg / kg / day or less, 20 mg / kg / day or less, 15 mg / kg / day or less, or 10 mg / kg / day or less, but is not limited thereto. The administration may be divided into one or more administrations per day. For example, administration may be performed 2 to 24 times a day, 1 to 2 times a day, 1 to 6 times a week, 1 to 10 times a week, 1 to 15 times a week, 1 to 3 times a week, 1 to 3 times a week, or 1 to 12 times a year, but is not limited thereto.

[0029] In an exemplary embodiment, the effective amount of at least one of the strains having the ability to produce phenyllactic acid; their lysates; their cultures; their fermentation products; and extracts of the strains, lysates, cultures or fermentation products when administered may be an amount such that the daily administration of phenyllactic acid reaches 0.1 to 100 mg / kg / day.

[0030] In an exemplary embodiment, when the active ingredient is a fermentation product of a strain having the ability to produce phenyllactic acid, the amount of the active ingredient applied may be 0.25 to 250 g / kg / day. More specifically, the amount of the active ingredient applied may be 0.25 g / kg / day or more, 0.5 g / kg / day or more, 0.75 g / kg / day or more, 1.0 g / kg / day or more, 1.25 g / kg / day or more, 1.5 g / kg / day or more, 1.75 g / kg / day or more, 2.0 g / kg / day or more, 2.25 g / kg / day or more, 2.5 g / kg / day or more, 2.75 g / kg / day or more, 3.0 g / kg / day or more, 3.25 g / kg / day or more, 3.5 g / kg / day or more, 3.75 g / kg / day or more, 4.0 g / kg / day or more, / day or more, 4.25g / kg / day or more, 4.5g / kg / day or more, 4.75g / kg / day or more, or 250g / kg / day or less, 200g / kg / day or less, 150g / kg / day or less, 100g / kg / day or less, 90g / kg / day or less, 80g / kg / day or less, 70g / kg / day or less, 60g / kg / day or less, 50g / kg / day or less, 40g / kg / day or less, 30g / kg / day or less, 20g / kg / day or less, or 10g / kg / day or less, but are not limited to these.

[0031] In the present application, "skin" includes mucous membranes. An example of a mucous membrane is the eye.

[0032] In the present application, the term "administering" refers to providing the composition according to the present invention to an administration subject by any appropriate method, and includes administration, coating, absorption and ingestion, etc. In this case, the administration subject refers to all animals to which the composition can be administered, such as humans, monkeys, dogs, goats, pigs or mice, etc.

[0033] In one exemplary embodiment, the composition can be administered to a subject in need of increasing expression of hyaluronan synthase in skin cells or eyes.

[0034] In an exemplary embodiment, the composition may be a transdermal composition or an oral composition.

[0035] In an exemplary embodiment, the composition may be at least one selected from a cosmetic composition for moisturizing or improving skin moisture, a quasi-drug composition, a general food composition, a health food composition, and a non-therapeutic oral composition.

[0036] In an exemplary embodiment, the composition may include 0.001 to 99.9% by weight of the active ingredient, for example, 0.01 to 20.0% by weight or 0.1 to 10.0% by weight of the active ingredient, but is not limited thereto.

[0037] In an exemplary embodiment, the cosmetic composition can be prepared in the form of a general emulsified formulation and a soluble formulation. The emulsified formulation includes nourishing lotion, cream, essence, etc., and the soluble formulation includes softening lotion, etc. Suitable formulations include, but are not limited to, for example, solutions, gels, solid or pasty anhydrous products, emulsions obtained by dispersing an oil phase in an aqueous phase, suspensions, microemulsions, microcapsules, tiny particle balls, ionic (liposomes) and non-ionic vesicle dispersants, creams, toners, lotions, powders, ointments, sprays or concealer sticks. In addition, it can include the form of foam or the form of an aerosol composition further comprising a compressed propellant.

[0038] In an exemplary embodiment, the cosmetic composition may further contain conventionally used adjuvants, such as fatty substances, organic solvents, solubilizers, concentrates, gelling agents, emollients, antioxidants, suspending agents, stabilizers, foaming agents, fragrances, surfactants, water, ionic or nonionic emulsifiers, fillers, metal ion chelators, chelating agents, preservatives, vitamins, retardants, humectants, necessary oils, dyes, pigments, hydrophilic or lipophilic active agents, lipid vesicles, or any other ingredients conventionally used in cosmetic compositions.

[0039] In an exemplary embodiment, when the composition is used as an additive to a general food or health food, it can be added directly or used together with other foods or food ingredients and can be used appropriately according to conventional methods. The amount of the active ingredient can be appropriately determined according to various purposes such as prevention, health care or treatment. The dosage form of the food can be powder, granules, pills, tablets, capsules, or any other form of general food or beverage.

[0040] In an exemplary embodiment, the types of general food or health food are not particularly limited. Examples of foods to which the composition can be added include meat, biscuits, noodles, chewing gum, dairy products including ice cream, various soups, drinking water, tea, beverages, alcoholic beverages and vitamin complexes, etc., and can include all foods in the conventional sense.

[0041] In an exemplary embodiment, the beverage in the general food or health food may contain various flavorings or natural carbohydrates as additional ingredients, just like ordinary beverages. The above-mentioned natural carbohydrates may be monosaccharides such as glucose and fructose; disaccharides such as maltose and sucrose; polysaccharides such as dextrin and cyclodextrin; and sugar alcohols such as xylitol, sorbitol, and erythritol. As sweeteners, natural sweeteners such as thaumatin and stevia extract can be used, or synthetic sweeteners such as saccharin and aspartame can be used. The proportion of the natural carbohydrates in each 100 mL of the beverage of the present invention may be about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g, but is not limited thereto.

[0042] In an exemplary embodiment, in addition to the above, general food or health food according to the present invention may include various nutrients, vitamins, electrolytes, flavor enhancers, colorants, pectin and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH regulators, stabilizers, preservatives, glycerol, alcohol, carbonating agents for carbonated beverages, etc. In addition, food according to the present invention may also contain the pulp used in the preparation of natural fruit juice, fruit juice beverages and vegetable beverages. These ingredients can be used alone or in combination. The ratio of these additives is not particularly limited, but under normal circumstances, based on 100 weight portions of health food according to the present invention, the content of these additives is selected from 0.01 weight portion to 0.1 weight portion.

[0043] In an exemplary embodiment, when the composition is used as a quasi-pharmaceutical additive, the composition can be added directly, or it can be used together with other quasi-pharmaceuticals or quasi-pharmaceutical ingredients, and can be used appropriately according to conventional methods. The mixing amount of the active ingredient can be appropriately determined according to the purpose of use. The ingredients contained in the quasi-pharmaceutical composition, in addition to the active ingredient, may also include ingredients conventionally used in quasi-pharmaceutical compositions, such as abrasives, wetting agents, adhesives, foaming agents, sweeteners, preservatives, medicinal ingredients, flavorings, pigments, solvents, whitening agents, solubilizers or pH regulators.

[0044] In another aspect, the present invention provides a method for preparing phenyllactic acid, comprising the following steps: culturing a Lactobacillus plantarum strain.

[0045] In an exemplary embodiment, the strain may be Lactobacillus plantarum APsulloc 331261 (Deposit No.: KCCM11179P).

[0046] In an exemplary embodiment, the method may include culturing the Lactobacillus plantarum strain in a culture medium supplemented with phenylalanine and / or phenylpyruvate.

[0047] In an exemplary embodiment, the method may involve culturing the Lactobacillus plantarum APsulloc 331261 (Deposit No.: KCCM11179P) strain in a culture medium supplemented with phenylalanine.

[0048] In an exemplary embodiment, the content of stupid alanine in the culture medium may be 1 mM or more and 150 mM or less, more specifically, the content may be 1 mM or more, 5 mM or more, 10 mM or more, 15 mM or more, 20 mM or more, 25 mM or more, 30 mM or more, 35 mM or more, 40 mM or more, 45 mM or more, 50 mM or more, 55 mM or more, 60 mM or more, 65 mM or more, 70 mM or more, 75 mM or more, or 150 mM or less, 145 mM or less, 140 mM or less, 135 mM or less, 130 mM or less, 125 mM or less, 120 mM or less, 115 mM or less, 110 mM or less, 105 mM or less, 100 mM or less, or 95 mM or less, but is not limited thereto. When the culture medium for culturing the strain contains 1 mM or more phenylalanine, phenyllactic acid can be efficiently produced. However, when the phenylalanine content in the culture medium exceeds 150 mM, it may have a negative impact on the growth of the strain and may reduce the production efficiency of phenyllactic acid.

[0049] In an exemplary embodiment, the culture medium may be a culture medium supplemented with calcium carbonate and used to culture the strain. By adding calcium carbonate to adjust the pH value of the culture medium, the production of phenyllactic acid can be increased.

[0050] In an exemplary embodiment, the pH of the culture medium may be 7.0 or higher, 7.1 or higher, 7.2 or higher, 7.3 or higher, 7.5 or higher, 7.6 or higher, 7.7 or higher, 7.8 or higher, 7.9 or higher, or 8.0 or higher, but is not limited thereto.

[0051] In an exemplary embodiment, the method may include culturing the Lactobacillus plantarum strain in a culture medium supplemented with phenylalanine and / or phenylpyruvate; and calcium carbonate.

[0052] In an exemplary embodiment, the method may include culturing Lactobacillus plantarum APsulloc 331261 (Deposit No.: KCCM11179P) in a culture medium supplemented with phenylalanine and calcium carbonate.

[0053] In yet another aspect, the present invention provides a method for moisturizing the skin or improving the moisture status of the skin, comprising applying to a subject an effective amount of a composition comprising at least one selected from phenyllactic acid; a strain having the ability to produce the phenyllactic acid; a lysate thereof; a culture thereof; a fermentation product thereof; and an extract of the strain, lysate, culture or fermentation product.

[0054] In yet another aspect, the present invention provides a method for promoting hyaluronic acid production, comprising administering to a subject an effective amount of a composition comprising at least one selected from the group consisting of phenyllactic acid; a strain having the ability to produce phenyllactic acid; a lysate thereof; a culture thereof; a fermentation product thereof; and an extract of the strain, lysate, culture or fermentation product.

[0055] In yet another aspect, the present invention provides a use of at least one of phenyllactic acid; a strain having the ability to produce phenyllactic acid; a lysate thereof; a culture thereof; a fermentation product thereof; and an extract of the strain, lysate, culture or fermentation product in preparing a composition for moisturizing the skin, a composition for improving the moisture status of the skin, or a composition for promoting the production of hyaluronic acid.

[0056] In yet another aspect, the present invention provides at least one of phenyllactic acid; a strain having the ability to produce phenyllactic acid; a lysate thereof; a culture thereof; a fermentation product thereof; and an extract of the strain, lysate, culture or fermentation product for moisturizing the skin, improving the moisture status of the skin, or promoting the production of hyaluronic acid.

[0057] In yet another aspect, the present invention provides non-therapeutic uses of at least one of phenyllactic acid; a strain having the ability to produce said phenyllactic acid; a lysate thereof; a culture thereof; a fermentation product thereof; and an extract of said strain, lysate, culture or fermentation product in moisturizing the skin, improving the moisture status of the skin, or promoting the production of hyaluronic acid.

[0058] Hereinafter, the constitution and effects of the present invention will be described in more detail with reference to examples. However, the purpose of providing these examples is only to help understand the present invention, and the category and scope of the present invention are not limited thereto.

[0059] [Experimental Example 1] Isolation of phenyllactic acid from various strains

[0060] This experimental example uses strains of various genera and isolates phenyllactic acid from them. Strains of various genera, including Lactobacillus acidophilus KCTC3164, Lactobacillus casei KCTC3109, Lactobacillus plantarum KCTC3108, Lactobacillus sakei KCTC3603, and Lactobacillus plantarum APsulloc331261 (KCCM11179P), which are strains of the genus Lactobacillus, were used to isolate phenyllactic acid. The above strains were inoculated into MRS broth and cultured at 35°C for 3 days. The culture was centrifuged at 1,800×g for 20 minutes at 4°C to separate the culture medium and the bacteria. The separated culture medium was filtered with a 0.2μm filter to completely remove the bacteria. The phenyllactic acid production of the culture medium prepared above was compared using HPLC analysis.

[0061] The results are shown in Table 1 and Figure 1 As shown, the phenyllactic acid production capacity of Lactobacillus plantarum strains is superior to that of other strains. In particular, the phenyllactic acid production of Lactobacillus plantarum APsulloc 331261 (KCCM11179P) strain is as high as approximately 100 ppm, while the phenyllactic acid production of Lactobacillus plantarum KCTC3108 strain is approximately 70.0 ppm. In contrast, the phenyllactic acid production of Lactobacillus acidophilus KCTC3164, Lactobacillus casei KCTC3109, and Lactobacillus salsa KCTC3603 strains is not detected, 22.8 ppm, and not detected, respectively. This shows that among strains of various genera, Lactobacillus plantarum, and particularly Lactobacillus plantarum APsulloc 331261 (KCCM11179P), has excellent phenyllactic acid production capacity. Therefore, in the following experimental examples, the Lactobacillus plantarum APsulloc 331261 (KCCM11179P) strain was used.

[0062]

Table 1

[0063] Comparison of phenyllactic acid production ability of strains from various genera

[0064]

[0065]

[0066]

[0067] [Experimental Example 2] Confirming the increase in phenyllactic acid production by Lactobacillus strains under different culture medium compositions and the addition of intermediate metabolites

[0068] The Lactobacillus plantarum APsulloc 331261 (KCCM11179P) strain used in the above experimental example was inoculated into a culture medium containing phenylalanine and phenylpyruvate added to MRS broth to confirm the production of phenyllactic acid. Specifically, 20, 40, or 80 mM of phenylalanine or phenylpyruvate was added to MRS culture medium, or calcium carbonate was added at a concentration of 1% above 40 mM of phenylalanine or phenylpyruvate, and the Lactobacillus strain was inoculated in the same manner. The culture was statically cultured at 35°C for 3 days, and then the production of phenyllactic acid was confirmed. In the same manner as in the above experimental example, the culture was centrifuged at 1,800 × g at 4°C for 20 minutes to separate the culture medium and the bacteria. The separated culture medium was filtered with a 0.2 μm filter to completely remove the bacteria. The culture medium prepared above was compared in terms of the amount of phenyllactic acid produced using HPLC analysis.

[0069] The results are as follows Figure 2 As shown, phenyllactic acid production increased with the addition of phenylalanine, reaching a maximum of 363.5 ppm when the pH was adjusted with calcium carbonate. Furthermore, Lactobacillus plantarum APsulloc331261 (KCCM11179P) failed to grow and produce phenyllactic acid when phenylpyruvic acid was added.

[0070] [Experimental Example 3] Isolation of Phenyllactic Acid from Lactobacillus Culture Medium

[0071] The Lactobacillus plantarum APsulloc 331261 (KCCM11179P) strain used in the above-mentioned experimental example was inoculated into a culture medium containing 40 mM phenylalanine and 1% calcium carbonate in MRS broth. The culture broth, which had been cultured for three days, was centrifuged to remove the bacterial cells, resulting in a 1-liter culture. An equal amount of ethyl acetate was added to the prepared culture broth to react, and the aqueous and ethyl acetate layers of the culture broth were allowed to separate. The separated ethyl acetate layer was heated and distilled, and an appropriate amount of water (approximately 100 mL) was added to the remaining concentrated solution to prepare a lactobacillus culture extract containing phenyllactic acid. This lactobacillus culture extract was confirmed to contain approximately 3000 ppm of phenyllactic acid.

[0072] [Experimental Example 4] Confirmation of the Promoting Effect of Phenyllactic Acid on Hyaluronic Acid Production

[0073] Human keratinocytes (HaCaT) were placed in a 35 mm culture dish and cultured in a 37°C and 5% CO2 incubator. After 24 hours, DMEM (containing gentamicin / amphotericin B, without FBS, Lonza) culture medium was treated with 50 μM (8.3 ppm) of PLA and the culture extract of Experimental Example 3. 0.5%, 0.25%, and 0.125% of the culture extract of Lactobacillus plantarum APsulloc 331261 (KCCM11179P) strain (the culture extract contains about 3000 ppm of PLA) were added and pretreated for 24 hours. After 24 hours of pretreatment, 1.25 ml of HBSS (Lonza) was added and the cells were irradiated with 410 nm blue light at 120 J / cm 2 The cells were irradiated with a dose of 50 μM PLA and 0.5%, 0.25%, and 0.125% PLA-containing lactobacillus culture extract were added to DMEM (containing gentamicin / amphotericin B, without FBS, Lonza) culture medium. The cells were cultured in a 37°C and 5% CO2 incubator for 24 hours. The cells were then collected, RNA was isolated, and cDNA was synthesized by RT-PCR (reverse transcriptional polymerase chain reaction). Taqman real-time quantitative PCR was performed using the synthesized cDNA to measure the gene expression of HAS2 (Hyaluronan synthase 2).

[0074] The results are as follows Figure 3 As shown, 50 μM PLA and 0.5%, 0.25%, and 0.125% PLA-containing lactobacillus culture extracts all showed significant improvement in the decrease in HAS2 gene expression induced by blue light.

[0075] In addition, the effects of the culture solutions of the Lactobacillus strains prepared as shown in Table 1 on promoting hyaluronic acid production were compared. Lactobacillus plantarum APsulloc 331261 (KCCM11179P), Lactobacillus plantarum KCTC3108, Lactobacillus acidophilus KCTC3164, and Lactobacillus sachetsii KTCT3603 were inoculated into MRS broth and cultured at 35°C for 3 days. The culture solution was added at 0.5%, and the gene expression level of HAS2 (hyaluronan synthase 2) was measured in the same manner as above.

[0076] The results are as follows Figure 4As shown, the culture fluid of Lactobacillus plantarum strain showed significant improvement on the decrease in HAS2 gene expression caused by blue light compared with the culture fluid of other Lactobacillus strains, especially the culture fluid of Lactobacillus plantarum APsulloc 331261 (KCCM11179P) strain was able to effectively improve the decrease in HAS2 gene expression.

[0077] The present description is further illustrated by the following embodiments which do not limit the claims.

[0078] Embodiment 1: selected from phenyllactic acid; a strain having the ability to produce phenyllactic acid; its lysate; its culture; its fermentation product; and the use of at least one of the extracts of the strain, lysate, culture or fermentation product in the preparation of a composition for moisturizing the skin, a composition for improving the moisture status of the skin, or a composition for promoting the production of hyaluronic acid.

[0079] Embodiment 2: The use according to embodiment 1, wherein the strain is a Lactobacillus plantarum strain.

[0080] Embodiment 3: The use according to any one of embodiments 1 to 2, wherein the strain is Lactobacillus plantarum APsulloc 331261 (Deposit No.: KCCM11179P).

[0081] Embodiment 4: The use according to any one of embodiments 1 to 3, wherein the strain converts phenylalanine and / or phenylpyruvate into phenyllactic acid.

[0082] Embodiment 5: The use according to any one of Embodiments 1 to 4, wherein the composition comprises 0.1 ppm to 1,000 ppm both inclusive of phenyllactic acid.

[0083] Embodiment 6: The use according to any one of embodiments 1 to 5, wherein the amount of phenyllactic acid administered is 0.1 to 100 mg / kg / day.

[0084] Embodiment 7: The use according to any one of embodiments 1 to 6, wherein the effective amount of at least one selected from the strain having the ability to produce phenyllactic acid; its lysate; its culture; its fermentation product; and the extract of the strain, lysate, culture or fermentation product when administered is an amount such that the daily administration of phenyllactic acid reaches 0.1 mg / kg / day to 100 mg / kg / day.

[0085] Embodiment 8: The use according to any one of embodiments 1 to 7, wherein when the active ingredient is a fermentation product of a strain capable of producing phenyllactic acid, the administration amount of the active ingredient is 0.25 g / kg / day to 250 g / kg / day.

[0086] Embodiment 9: The use according to any one of embodiments 1 to 8, wherein the composition is administered to a subject in need of increasing the expression of hyaluronan synthase in skin cells.

[0087] Embodiment 10: The use according to any one of embodiments 1 to 9, wherein the composition is a transdermal composition or an oral composition.

[0088] Embodiment 11: The use according to any one of Embodiments 1 to 10, wherein the composition is a cosmetic composition or a quasi-drug composition.

[0089] Embodiment 12: The use according to any one of Embodiments 1 to 10, wherein the composition is a general food composition or a health food composition for improving skin moisture status.

[0090] Embodiment 13: A method for preparing phenyllactic acid, comprising the following steps: culturing a Lactobacillus plantarum strain.

[0091] Embodiment 14: The method according to embodiment 13, wherein the strain is Lactobacillus plantarum APsulloc331261 (deposit number: KCCM11179P).

[0092] Embodiment 15: The method according to any one of embodiments 13 to 14, wherein the method comprises culturing the Lactobacillus plantarum strain in a culture medium supplemented with phenylalanine and / or phenylpyruvate; and / or calcium carbonate.

[0093] Embodiment 16: The method according to any one of Embodiments 13 to 15, wherein the method comprises culturing the Lactobacillus plantarum APsulloc 331261 (Deposit No.: KCCM11179P) strain in a culture medium supplemented with styrylamine and / or calcium carbonate.

[0094]

Collection Information

[0095] Name of depository institution: Korea Collection of Microorganisms (KCCM);

[0096] Accession number: KCCM11179P;

[0097] Date of deposit: March 28, 2011.

Claims

1. selected from phenyllactic acid; strains capable of producing said phenyllactic acid; and lysates thereof; its culture; its fermentation products; And use of at least one of the extracts of the strain, lysate, culture or fermentation product in preparing a composition for moisturizing the skin, a composition for improving the moisture status of the skin, or a composition for promoting the production of hyaluronic acid.

2. The use according to claim 1, wherein The strain is a Lactobacillus plantarum strain.

3. The use according to claim 1, wherein The strain is Lactobacillus plantarum APsulloc 331261, and its deposit number is KCCM11179P.

4. The use according to claim 1, wherein The strain converts phenylalanine and / or phenylpyruvate into phenyllactic acid.

5. The use according to claim 1, wherein The composition contains phenyllactic acid in an amount of 0.1 ppm or more and 1,000 ppm or less.

6. The use according to claim 1, wherein The daily administration amount of phenyllactic acid is 0.1 mg / kg / day to 100 mg / kg / day.

7. The use according to claim 1, wherein Selected from strains having the ability to produce phenyllactic acid; lysates thereof; its culture; its fermentation products; The effective amount of at least one of the extracts of the strain, lysate, culture or fermentation product when administered is an amount such that the daily administration of phenyllactic acid reaches 0.1 mg / kg / day to 100 mg / kg / day.

8. The use according to claim 1, wherein When the active ingredient is a fermentation product of a strain having the ability to produce phenyllactic acid, the administration amount of the active ingredient is 0.25 g / kg / day to 250 g / kg / day.

9. The use according to claim 1, wherein The composition is administered to a subject in need of increasing the expression of hyaluronan synthase in skin cells.

10. The use according to claim 1, wherein The composition is a transdermal composition or an oral composition.

11. The use according to claim 1, wherein The composition is a cosmetic composition or a quasi-pharmaceutical composition.

12. The use according to claim 1, wherein The composition is a general food composition or a health food composition for improving the moisture state of the skin.

13. A method for preparing phenyllactic acid, comprising the following steps: culturing a Lactobacillus plantarum strain.

14. The method according to claim 13, wherein The strain is Lactobacillus plantarum APsulloc 331261 strain, and its preservation number is KCCM11179P.

15. The method according to claim 13, wherein The Lactobacillus plantarum strain is cultured in a culture medium supplemented with phenylalanine and / or phenylpyruvate; and / or calcium carbonate.

16. The method according to claim 13, wherein: The Lactobacillus plantarum APsulloc 331261 strain with the deposit number KCCM11179P was cultured in a culture medium supplemented with phenylalanine and / or calcium carbonate.

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