阅读说明：本技术 包含新型鼠李糖乳杆菌lm1011作为有效成分的皮肤美白用化妆品组合物 (Cosmetic composition for skin whitening comprising novel lactobacillus rhamnosus LM1011 as active ingredient ) 是由 全珉圭 李智燕 洪度先 于 2020-04-24 设计创作，主要内容包括：本申请涉及一种包含新型鼠李糖乳杆菌LM1011作为有效成分的皮肤美白用化妆品组合物,更详细地,提供一种通过调整酪氨酸酶活性来降低黑色素生成的包含新型鼠李糖乳杆菌LM1011作为有效成分的皮肤美白用化妆品组合物。(The present invention relates to a skin whitening cosmetic composition comprising a novel lactobacillus rhamnosus LM1011 as an active ingredient, and more particularly, to a skin whitening cosmetic composition comprising a novel lactobacillus rhamnosus LM1011 as an active ingredient, which reduces melanin production by regulating tyrosinase activity.)

1. A cosmetic composition for skin whitening, characterized by comprising a novel Lactobacillus rhamnosus LM1011 as an active ingredient.

2. The cosmetic composition for skin whitening according to claim 1, wherein the cosmetic composition for skin whitening is contained at a concentration of 1 x 10¹¹cells/mL to 5X 10¹¹cells/mL of the lactobacillus rhamnosus LM 1011.

3. The cosmetic composition for skin whitening according to claim 1, wherein the lactobacillus rhamnosus LM1011 is capable of reducing melanin production.

4. The cosmetic composition for skin whitening according to claim 3, wherein the Lactobacillus rhamnosus LM1011 reduces melanin production by modulating tyrosinase activity.

5. The cosmetic composition for skin whitening according to claim 1, wherein the cosmetic composition for skin whitening is in a dosage form selected from the group consisting of: skin lotions, emulsions, essences, creams, masks, foundations, and sun screens.

Technical Field

The present invention relates to a skin whitening cosmetic composition containing a novel lactobacillus rhamnosus LM1011 as an active ingredient, and more particularly, to a skin whitening cosmetic composition containing a novel lactobacillus rhamnosus LM1011 as an active ingredient, which reduces melanin production by regulating tyrosinase activity.

Background

Human skin color is largely dependent on the amount of pigment known as melanin (melanin) present in the skin. Melanin is biosynthesized by melanocytes (melanocytes) present in the basal layer of the skin, and moves from the basal layer of the epidermis to the horny layer by the keratinization process of keratinocytes (keratinocytes) under the action of cytoplasmic processes.

In melanogenic cells, tyrosine (tyrosine), which is an amino acid, is converted into DOPA (DOPA), dopaquinone (DOPA quinone) by an enzyme called tyrosinase (tyrosine), and then melanin is produced through a plurality of oxidation processes. This melanin pigment absorbs ultraviolet rays and protects various cell tissues present in the epidermis from the ultraviolet rays, and if it is synthesized excessively, it not only causes spots and freckles but also darkens the whole skin. Therefore, if excessive melanin synthesis can be inhibited, not only the skin color can be lightened, but also the skin pigmentation symptoms of spots and freckles can be alleviated.

On the other hand, Lactobacillus rhamnosus (Lactobacillus rhamnosus) is a lactic acid bacterium, and is known to have various effects such as intestinal regulation (intestinal regulation), antibacterial action, prevention of allergy, reduction of body fat, and the like. In addition, a great deal of research has been conducted in recent years on sterilized lactic acid bacteria, and it has been found that Lipoteichoic acid (Lipoteichoic acid), which is one of cell wall components generated in the process of killing live bacteria, adheres to the barrier to inhibit the adhesion of harmful bacteria, and that antibacterial components in fermentation products such as lactic acid (lactic acid) and bacteriocin (bacteriocin) directly act to inhibit harmful bacteria, due to the action of the sterilized lactic acid bacteria. In addition, it has been found that immunoglobulin a, which is an important substance for protection against infection by external bacteria and the like, is produced by stimulating macrophages by absorption of a sterilized lactic acid bacterium in the small intestine, as compared with the living bacteria of a normal lactic acid bacterium. It has also been found that the sterilized lactic acid bacteria are used as food for live bacteria, so that the sterilized lactic acid bacteria can play an excellent role of Prebiotics (probiotics) in intestinal regulation (intestinal regulation) of lactic acid bacteria.

Disclosure of Invention

Problems to be solved by the invention

Provided is a skin whitening cosmetic composition that reduces melanin production by regulating tyrosinase activity and contains a novel Lactobacillus rhamnosus LM1011 as an active ingredient.

However, the problems to be solved by the present application are not limited to the above-mentioned problems, and other problems not mentioned can be clearly understood by those skilled in the art through the following description.

Means for solving the problems

A first aspect of the present application provides a cosmetic composition for skin whitening comprising Lactobacillus rhamnosus LM1011(Lactobacillus rhamnosus LM1011, depository: Korean type culture Collection, accession No: KCTC12751BP, depository date: 2019, 4/18 days).

Effects of the invention

A skin whitening cosmetic composition according to an embodiment of the present application, which contains Lactobacillus rhamnosus LM1011(Lactobacillus rhamnosus LM1011, depository: Korean type culture Collection, accession No: KCTC12751BP, depository date: 2019, 4/18 days), as an active ingredient, has the effect of whitening the skin by decreasing melanin production by regulating tyrosinase activity, and particularly has the whitening effect on hypermelanosis (hypermelaniss).

Drawings

FIG. 1 is a schematic diagram showing a mechanism of reducing melanin production by Lactobacillus rhamnosus LM1011(Lactobacillus rhamnosus LM1011, depository: Korean type culture Collection, accession No. KCTC12751BP, depository date: 2019, 4/18) in the examples of the present application.

Fig. 2 is a graph showing a melanin production reducing effect of a whitening cosmetic composition comprising lactobacillus rhamnosus LM1011 in an example of the present application in a B16-F10 melanoma cell line.

FIG. 3 is a graph illustrating the use of a whitening cosmetic composition comprising Lactobacillus rhamnosus LM1011 in a clinical trial of whitening effect against hypermelanosis (hypermelanosis) in an example of the present applicationGraph of the measured improvement rate of M-value (melanin index) according to the change of time.

Fig. 4 is a graph showing a change of a visual evaluation score of a whitening cosmetic composition comprising lactobacillus rhamnosus LM1011 according to time in a clinical trial of a whitening effect against hypermelanosis (hypermelaniss) in an example of the present application.

Fig. 5 is a graph showing the change of the improvement rate of the L-value (lightness value) measured using a colorimeter (Chromameter) according to time in a clinical trial of the whitening effect against hypermelanosis (hypermelaniss) in the embodiment of the present application.

Fig. 6 is a graph showing whether lactobacillus rhamnosus LM1011 has cytotoxicity in the examples of the present application.

FIG. 7 is a graph showing whether Lactobacillus rhamnosus HK-9 has cytotoxicity in the examples of the present application.

FIG. 8 is a graph showing the effect of Lactobacillus rhamnosus LM1011 and Lactobacillus rhamnosus HK-9 in inhibiting Tyrosinase (Tyrosinase) expression upon melanogenesis promoting hormone (α -MSH) treatment in an embodiment of the present application.

Detailed Description

Hereinafter, embodiments of the present application will be described in detail with reference to the accompanying drawings so as to be easily implemented by those skilled in the art described in the present application. However, the present application may be embodied in many different forms and is not limited to the embodiments described herein. In addition, in order to more clearly explain the present application in the drawings, portions that are not related to the description are omitted, and like reference numerals are given to like portions throughout the specification.

Throughout the specification, when a certain portion is referred to as being "connected" to another portion, the term includes not only a case of "directly connecting" but also a case of "indirectly connecting" with a substance such as a linker interposed therebetween.

Throughout the specification of the present application, when one member is "on" another member, not only a case where the member is in contact with the other member but also a case where the other member is present between the two members is included.

Throughout the specification, when a certain portion is referred to as "including" a certain structural element, unless explicitly stated to the contrary, it is to be understood that the other structural element is not excluded, and the other structural element may be included. The terms "about," "substantially," and the like, as used throughout the specification are used in the sense of their data or close to their data in the sense of suggesting inherent manufacturing and material tolerances in the meaning referred to, in order to prevent others from maliciously and inappropriately utilizing the disclosure herein which gives rise to accurate or absolute numerical values for ease of understanding. The term "step of performing" or "step of" used throughout the specification of the present application does not mean "step for".

Throughout the specification of the present application, the term "combination(s)" included in the expression of markush system means a mixture or combination of one or more selected from the group consisting of the structural elements described in the expression of markush system, in other words, means one or more selected from the group consisting of the structural elements described above.

Throughout the specification of the present application, the expression "a and/or B" means "a or B, or a and B".

Hereinafter, embodiments and examples of the present application will be described in detail with reference to the accompanying drawings. However, the present application is not limited to these embodiments and examples and drawings.

The first embodiment of the present application provides a skin whitening cosmetic composition comprising Lactobacillus rhamnosus LM1011(Lactobacillus rhamnosus LM1011, depository: korean typical culture collection, accession No. KCTC12751BP, depository date: 2019, 4/18 days).

In one embodiment of the present application, the skin whitening cosmetic composition may include the lactobacillus rhamnosus LM1011 strain, dead cells thereof, broken cells, culture products, concentrates, or extracts thereof as an active ingredient, but is not limited thereto. For example, the above-mentioned cosmetic composition for skin whitening may include a heat-treated lactobacillus rhamnosus LM1011 strain as an effective ingredient, but is not limited thereto.

In an embodiment of the present application, the lactobacillus rhamnosus LM1011 strain described above may be heat-treated to utilize the dead cells of lactic acid bacteria themselves, and when the lactobacillus rhamnosus LM1011 strain described above is applied to a cosmetic product, product instability due to abnormal fermentation may be reduced, but is not limited thereto.

In one embodiment of the present application, when the skin whitening cosmetic composition includes lactobacillus rhamnosus LM1011, the content thereof is not particularly limited, and for example, the lactobacillus rhamnosus LM1011 may be included at a concentration of about 1 × 10¹¹cells/mL to about 5X 10¹¹cells/mL, but not limited thereto.

In one embodiment of the present application, when the skin whitening cosmetic composition includes the lactobacillus rhamnosus LM1011 at a concentration of less than about 1 × 10¹¹cells/mL or greater than about 5X 10¹¹The skin whitening effect may be reduced at a cell/mL, and thus the concentration of Lactobacillus rhamnosus LM1011 may be about 1X 10¹¹cells/mL to about 5X 10¹¹cell/mL, about 1X 10¹¹cells/mL to about 4X 10¹¹cell/mL, about 1X 10¹¹cells/mL to about 3X 10¹¹cell/mL, about 1X 10¹¹cells/mL to about 2X 10¹¹cell/mL, about 2X 10¹¹cells/mL to about 5X 10¹¹cells/mL orAbout 3X 10¹¹cells/mL to about 5X 10¹¹cells/mL, but not limited thereto.

In one embodiment of the present application, the lactobacillus rhamnosus LM1011 can reduce the production of melanin, but is not limited thereto. For example, the lactobacillus rhamnosus LM1011 described above can reduce the production of melanin in the skin, which is increased due to exposure to ultraviolet rays, but is not limited thereto.

In one embodiment of the present application, the lactobacillus rhamnosus LM1011 can reduce melanin production by regulating tyrosinase activity, but is not limited thereto. For example, the lactobacillus rhamnosus LM1011 changes L-tyrosine into L-dopa, and modulates tyrosinase activity of an enzyme for producing melanin by changing the L-dopa into dopaquinone, and particularly, reduces the production of melanin in the skin by inhibiting the expression level and/or activity of tyrosinase, but is not limited thereto.

In one embodiment of the present application, the formulation of the above-mentioned skin whitening cosmetic composition is not particularly limited, and for example, may be selected from any one of lotions, emulsions, essences, creams, masks, foundations, and barrier creams, but is not limited thereto.

In an embodiment of the present application, the skin whitening cosmetic composition may be added with a material arbitrarily selected according to a formulation form or a use purpose of the cosmetic, but is not limited thereto. For example, purified water, oil, surfactant, humectant, higher alcohol, thickener, chelating agent, pigment, fatty acid, antioxidant, preservative, wax, pH adjuster, perfume, etc., may be added, but not limited thereto.

In one conventional example of the present application, when the above-mentioned skin whitening cosmetic composition is in the form of a paste, cream or gel, the carrier (carrier) component may include: animal oil, vegetable oil, wax, paraffin, starch, amine yellow gum, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, zinc oxide, or the like, but is not limited thereto.

In an embodiment of the present application, when the skin whitening cosmetic composition is in the form of powder or spray, the carrier component may include: lactose, talc, silicon dioxide, aluminium hydroxide, calcium silicate or polyamide powder, especially in the case of spraying, may additionally comprise propellants, such as chlorofluorocarbons, propane/butane or dimethyl ether, without being limited thereto.

In an embodiment of the present application, when the above-mentioned cosmetic composition for skin whitening is in the form of a solution or emulsion, the carrier component may include: solvents, solubilizers or emulsifiers, for example: water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1, 3-butyl glycol oil, glycerol fatty ester, polyethylene glycol, or fatty acid ester of sorbitan, but is not limited thereto.

In an embodiment of the present application, when the above-mentioned cosmetic composition for skin whitening is in the form of a suspension, the carrier component may include: liquid diluents, for example: water, ethanol or propylene glycol; suspensions, for example: ethoxylated isostearyl alcohols, polyoxyethylene sorbitol esters, and polyoxyethylene sorbitan esters; microcrystalline cellulose; aluminum methylhydroxide, bentonite, agar, or amine tragacanth, and the like, but are not limited thereto.

In an embodiment of the present application, when the skin whitening cosmetic composition is in the form of a surfactant-containing cleanser, the carrier component may include: aliphatic alcohol sulfates, aliphatic alcohol ether sulfates, sulfosuccinic acid monoesters, isethionates, imidazoline derivatives, methyltaurates, sarcosinates, fatty acid amide ether sulfates, alkylamidobetaines, fatty alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives, ethoxylated glycerol fatty acid esters, or the like, but are not limited thereto.

A second embodiment of the present application provides a method for improving skin comprising the step of applying/fitting a skin whitening cosmetic composition comprising Lactobacillus rhamnosus LM1011(Lactobacillus rhamnosus LM1011, depository: korean typical culture depository, accession No. KCTC12751BP, depository date: 2019, 4/18 days). Note that the same contents as those of the first embodiment are also applicable to the second embodiment.

Best mode for carrying out the invention

Hereinafter, the present application will be described in more detail by examples thereof, but the following examples are merely illustrative to facilitate understanding of the present application, and the contents of the present application are not limited to the following examples.

1. Production of dead cells of lactic acid bacteria and cell test

1-1 preparation of cosmetic composition for skin whitening comprising Lactobacillus rhamnosus LM1011

The dead cells of Lactobacillus rhamnosus LM1011 according to this example were prepared by a batch culture method.

The cultivation of the above lactic acid bacteria was carried out in a 100L fermenter (Cobiotech) at a stirring speed of 30rpm at a temperature of 35 ℃ to 37 ℃ and a pH of 5 to 6.5 in a total amount of 60L of the culture solution. After the culture, the culture broth was centrifuged at 13000rpm for 30 minutes to collect the cells. The recovered cells were diluted 2-fold by weight with physiological saline (0.9% NaCl), and then subjected to tyndall at 110 ℃ for 10 minutes in an autoclave, and freeze-dried to finally obtain heat-treated dead cells of lactic acid bacteria (Lactobacillus rhamnosus LM 1011).

1-2 melanin production-reducing effect (in vitro)

The effect of reducing melanin production was confirmed for the heat-treated lactobacillus rhamnosus LM1011 obtained in the above example. The B16-F10 melanoma cell line was used as a cell line and was plated at 1X 10 in a 6-well cell culture plate (6-well cell culture plate)⁴Concentration of cells/well (cells/well). After 24 hours of culture, the concentrations of heat-treated Lactobacillus rhamnosus LM1011 in the above cell lines were adjusted to 0 cells/mL (negative control group) and 1X 10 cells/mL, respectively⁴cell/mL, 1X 10⁵cell/mL, 1X 10⁶cell/mL, 1X 10⁷cell/mL and 1X 10⁸cells/mL, and cultured for 48 hours. EX-CYTOX (EZ-3000, Daeil lab service) may be used and read by a Multi-plate reader (Multi-plate reader)The amount of protein expressed in the cells was measured by measuring absorbance at 450nm, and then whether or not the melanin production inhibitory effect was exhibited was determined by comparing the melanin synthesis inhibitory rate with respect to the negative control group. In addition to the above-mentioned heat-treated lactobacillus rhamnosus LM1011 strain experimental group, a compound β -arbutin showing whitening function was also compared as a 200ug/mL positive control group.

As shown in FIG. 2, the heat-treated Lactobacillus rhamnosus LM1011 was at 1X 10 compared to the negative control group⁵cells/mL to 1X 10⁸The cells/mL showed potent melanin production-reducing effect, particularly at 1X 10⁸The cell/mL concentration showed a melanin production-reducing effect of about 20% higher than that of beta-arbutin.

2. Production of cosmetics containing dead cells and suitability test for human body

2-1. cosmetic preparation

The dead lactobacillus rhamnosus LM1011 strain prepared in the above examples is diluted to the weight ratio of 2 to 10 times, and the cell concentration is adjusted to 1 × 10¹¹cells/mL.

Control and experimental group essences (serum) were prepared by a conventional method according to the compounding amounts shown in table 1 below.

[ TABLE 1 ]

Composition (I)	Compounding amount (%)	Control group	Experimental group
				Dead bacteria	3	X	○
Polyethylene glycol	8	○	○
				Ethyl hexyl glycerol	0.1	○	○
Butanediol	0.25	○	○
				Purified water	Residual amount of	○	○

2-2 whitening effect on hypermelanosis (hypermelaniss) (in vivo vivo))

The whitening effect of the skin whitening cosmetic composition comprising the heat-treated lactobacillus rhamnosus LM1011 as an active ingredient obtained in the above examples was confirmed by clinical experiments. The experiment was performed on 23 hypermelanotic patients who met the selection criteria and did not incorporate the exemption criteria, and a method of self-application twice daily was applied. By making use ofImprovement of M-value (pigmentation)Index), evaluation of the subject, evaluation of skin irritation by a dermatologist, and visual evaluation by a dermatologist, etc. confirm the whitening effect.

As shown in FIG. 3, the difference between the M-value before the test and the M-value after the test was obtained at each of the control part and the test part, thereby obtaining the pigmentation improvement rate of the cosmetic composition for skin whitening. The results represent the improvement over time by expressing it as a percentage of the measured value before the test. The degree of reduction of pigmentation after the skin whitening cosmetic composition of the present invention was applied on the skin increased with the increase of the application time, compared to the placebo group, which was the control group.

As shown in fig. 4, the visual evaluation score of the skin whitening cosmetic composition was obtained from the difference between the control part and the test part before and after the test. The skin whitening cosmetic composition of the present application increases the degree of pigmentation reduction with the administration time, compared to the placebo group, which is the control group.

The above results indicate that the skin whitening cosmetic composition comprising the heat-treated lactobacillus rhamnosus LM1011 of the present application as an effective ingredient has melanin reduction and accompanying skin whitening effects when applied to the skin.

2-3 whitening effect (in vivo)

The whitening effect of the skin whitening cosmetic composition comprising the heat-treated lactobacillus rhamnosus LM1011 as an active ingredient obtained in the above examples was confirmed by clinical experiments. The experiment was performed on 23 hypermelanotic patients who met the selection criteria and did not incorporate the exemption criteria, and a method of self-application twice daily was applied. The whitening effect was expressed as the improvement rate of the skin brightness index by measuring the brightness value (L-value) of the face area using a colorimeter (Chromameter).

As shown in fig. 5, the whitening effect of the cosmetic composition for skin whitening was obtained by obtaining the difference between the L value before the test and the L value after the test at the control site and the test site, respectively. The results represent the improvement over time by expressing it as a percentage of the measured value before the test. The cosmetic composition for skin whitening of the present application significantly improved the improvement rate of skin brightness index after use, compared to the placebo group as a control group. The above results indicate that the skin whitening cosmetic composition comprising the heat-treated lactobacillus rhamnosus LM1011 of the present application as an effective ingredient has a whitening effect when applied to the skin.

3. Evaluation of tyrosinase (tyrosinase) inhibitory Activity of Lactobacillus rhamnosus LM1011

3-1. confirmation of cytotoxicity

In order to compare and confirm the inhibitory activity of Lactobacillus rhamnosus HK-9 (hereinafter, HK-9) and tyrosinase (tyrosinase), which is a key enzyme regulating the melanin production reaction of Lactobacillus rhamnosus LM1011, the following experiment was performed.

First, in order to confirm the cytotoxicity of LM1011 and HK-9 and to determine the treatment concentration, B16F10 cells were treated at 5X 10⁴One/well was seeded in 96-well plates. After 24 hours of stabilization, 1X 10²To 1X 10⁸Lactic acid bacteria per ml were treated. After 24 hours, 95ul of DMEM (gibco) and 5ul of CCK-8 kit (Cell Counting kit-8) (CCK-8, Woongbe Meditech) were mixed and processed. After 2 hours, and measured by a Multi-plate reader (450nm), and converted to percentages based on the control group, and cytotoxicity was compared.

As a result, when the results shown in FIGS. 6 and 7 were observed, it was confirmed that there was no statistical difference in cytotoxicity according to the treatment concentrations of LM1011 and HK-9, and in the next experiment, the treatment concentrations of LM1011 and HK-9 were determined to be 1X 10⁸One per ml.

3-2 evaluation of inhibition of tyrosinase expression

In order to evaluate the inhibition effect of alpha-melanogenesis stimulating hormone (alpha-MSH) treatment on tyrosinase expression according to Lactobacillus rhamnosus LM1011 and Lactobacillus rhamnosus HK-9, the following experiments were performed.

First, the B16F10 cell line was cultured at 2X 10⁵One/holeInoculate in 6-well plates. After 24 hours of stabilization, the experiments were divided into an untreated group (Nor), an α -MSH-treated group, an α -MSH + HK-9-treated group, and an α -MSH + LM 1011-treated group. After 24 hours of treatment, the medium was removed, washed with PBS, and then treated with 500ul Trizol to destroy the cells. After sufficient cell disruption, the cells were treated with 100ul of chloroform (chloroform) and centrifuged (13000rpm, 15 minutes, 4 ℃) to separate cell debris (cell debris) and RNA. After taking 240ul of the supernatant, RNA was precipitated by treatment with an equal amount of Isopropanol (IPA) and centrifuged (13000rpm, 15 min, 4 ℃). The supernatant was removed and treated with 700ul of 75% ethanol (13000rpm, 5 min, 4 ℃) and centrifuged. After drying sufficiently, the RNA was dissolved in 30ul of ultrapure water (Ultra pure water).

Next, after the isolated RNA was quantified and 1. mu.g of the RNA was collected, cDNA synthesis was performed using an iScript cDNA synthesis kit (iScript cDNAsynthesis kit) (BIO-RAD). The PCR reaction was performed as shown in Table 2 below.

[ TABLE 2 ]

Reaction step	Reaction conditions
		Starting up	At 25 ℃ for 5 minutes
Reverse transcription	At 46 ℃ for 20 minutes
		Inactivation of RT	At 95 ℃ for 1 minute
Optional steps	Keeping at 4 deg.C

After completion of the cDNA synthesis, the expression level of Tyrosinase was evaluated by Real-time PCR (viii 7, Applied Biosystems) using a TaqMan probe (TaqMan probe) (Tyrosinase, Mm00495817_ m1, Applied Biosystems) using 260ng as a template (template). As a result, it was confirmed that the expression level of tyrosinase was significantly reduced in the LM 1011-treated group as compared with the HK-9-treated group under the stimulation of α -MSH (FIG. 8, p < 0.05).

From the above results, it was confirmed that the LM1011 of the present invention can more remarkably inhibit the expression of tyrosinase stimulated by a melanogenesis-promoting hormone than the same Lactobacillus rhamnosus strain HK-9 at a concentration without cytotoxicity, and it is considered that the LM1011 of the present invention has a remarkably superior inhibitory effect on the synthesis of melanin in the skin than the existing Lactobacillus rhamnosus strain. Therefore, the LM1011 strain of the present invention can be used as a composition for improving whitening, and is suitable for functional materials of cosmetics, foods, and the like.

The above description is for illustrative purposes of the present application, and it should be understood by those of ordinary skill in the art to which the present application pertains that the present application may be readily converted into other specific forms without changing the technical spirit or essential features of the present application. It is therefore to be understood that the above embodiments are illustrative in all respects and not restrictive. For example, each constituent element described as a single type may be implemented in a distributed manner, and similarly, constituent elements described as distributed may also be implemented in a combined form.

The scope of protection of the present application is indicated by the following claims in comparison with the detailed description made above, and all changes or modifications derived from the meaning and scope of the claims and equivalent concepts should be construed as being included in the scope of protection of the present application.

[ DENSATION NUMBER ]

The preservation organization: korean type culture Collection

The preservation number is as follows: KCTC12751BP

The preservation date is as follows: 18/4/2019

PCT/RO/134 Table