KR101760766B1 - Novel Lactobacillus curvatus GFC-AJ6, Lactic Acid Bacteria Fermented Product Comprising Polyamines Using Lactobacillus curvatus GFC-AJ6 and Producing Method Thereof - Google Patents

Abstract

The present invention relates to a novel Lactobacillus carbatus GFC-AJ6 strain, a fermentation product of a polyamine-containing lactic acid bacterium using the strain, and a process for producing the same.
A polyamine produced through bioconversion from arginine as a substrate using the novel Lactobacillus curvatus GFC-AJ6 strain (Accession No: KCCM11932P) provided from the present invention Since fermented products of lactic acid bacteria containing Putrescine, Spermidine and Spermine are produced, skin whitening or wrinkle-reducing effects are excellent. Therefore, the fermented product of polyamine-containing lactic acid bacteria as an active ingredient When applied to a cosmetic base, has an excellent effect as a cosmetic composition.

Description

[0001] The present invention relates to a novel Lactobacillus carbatus GFC-AJ6 strain, a fermentation product of a polyamine-containing lactic acid bacterium using the strain, and a method for producing the same. }

The present invention relates to a novel Lactobacillus curvatus GFC-AJ6 strain, a fermentation product of a polyamine-containing lactic acid bacterium using the strain, and a process for producing the same.

Polyamine is a generic term for substances with two or more amino groups synthesized from amino acids in the body. It is synthesized vigorously in the growth phase in all cells, plants and human cells, and exists in animals, plants and microorganisms. (Liu, Ran, et al., Talanta. 2011).

In addition, polyamines in foods and beverages are produced by the enzymatic action of raw materials and the decanoic acid action of amino acids in microorganisms. Among these amino acids, putrescine, spermidine and spermine, which are among the polyamines, are indispensable components of all living cells (Romain et al., Pediatric Res 1992; Bardoocz et al., J. Nutr. Biochem., 1993). These polyamines are essential components for maintaining the function of the intestines and the metabolism of the immune system.

As described above, representative examples of the polyamines are three kinds of fu- tresin, spermidine and spermine. Futresine is derived from arginine and ornithine to become a precursor of spermidine, and spermine is derived from spermidine (Coleman, C. S, et al., Biochemical Journal, 2004). In addition, polyamines exist as cations at the pH of the body and can react electrically with anionic species such as DNA, RNA, acids, phospholipids, and proteins. Therefore, it is known as an essential factor for cell growth and proliferation and membrane stability by stabilizing and regulating nucleic acid (Pegg, A. E, IUBMB life. 2009).

Production of polyamines has been reported to be produced by various pathways in prokaryotes and eukaryotes, and has been found in many meat and fermented foods (Iyer, Ramaswamy K. et al., Molecular genetics and metabolism 2002).

Further, the microorganism to produce a polyamine is Bacillus (Bacillus), Lactobacillus bacteria (Lactobacillus), Phedi O Rhodococcus (Pediococcus), Streptococcus (Streptococcus) , Which are microorganisms present in various fermented foods.

Recently, polyamines have attracted attention as a cosmetic material that imparts beneficial effects. However, the technology for using polyamines produced by microorganisms is disclosed in Korean Patent Registration No. 10-1485970, " Novel Lactobacillus sp. AML 2-1 strain, A method for producing polyamines and gamma aminobutyric acid using the same, and use of polyamines and gamma aminobutyric acid ", Korean Patent Registration No. 10-1373224" A novel Lactobacillus sp. AML 15 strain producing polyamines and a culture solution thereof as an active ingredient &Quot;, and research on the functional properties and physiological activities of polyamines in skin cells has been lacking.

Accordingly, the present inventors selected Lactobacillus curvatus GFC-AJ6 (Accession No .: KCCM11932P), which is excellent in the ability to produce polyamines separated from Korean traditional fermented food turnip kimchi, Synthesis pathway When producing fermented lactic acid bacteria containing Putrescine, Spermidine and Spermine, which are polyamines produced by bioconversion from the early precursor arginine, Skin whitening or wrinkle-improving effect can be imparted, and the present invention has been completed.

Korean Patent Publication No. 10-1485970 Korean Patent Publication No. 10-1373224

It is an object of the present invention to provide a Lactobacillus curvatus GFC-AJ6 strain (accession number: KCCM11932P) having polyamine production ability.

Another object of the present invention is to provide a method for producing polyamine-containing polyamines, which comprises culturing a strain of Lactobacillus curvatus GFC-AJ6 (accession number: KCCM11932P) in a medium to which arginine is added And a method for producing a fermented product of lactic acid bacteria.

It is still another object of the present invention to provide a cosmetic composition for skin whitening or wrinkle improvement containing the polyamine-containing fermented lactic acid bacteria as an active ingredient.

In order to achieve the above object, the present invention provides a strain of Lactobacillus curvatus GFC-AJ6 (Accession No .: KCCM11932P) having polyamine production ability.

In the present invention, the strain is characterized by being derived from a turnip kimchi.

In the present invention, the polyamines may be at least one selected from the group consisting of putrescine, spermidine and spermine.

The present invention also relates to a method for producing a polyamine-containing lactic acid bacterium, which comprises culturing a strain of Lactobacillus curvatus GFC-AJ6 (accession number: KCCM11932P) in a medium to which arginine is added Thereby producing a fermented product.

In the present invention, the medium is an MRS medium containing 0.5 to 1.0% by weight of arginine.

In the present invention, the culturing is performed at 30 to 37 DEG C for 3 to 5 days.

The present invention also provides a fermented product of a polyamine-containing lactic acid bacterium produced through the above-described production method.

The present invention also provides a cosmetic composition for skin whitening comprising the fermented product of lactic acid bacteria containing polyamines as an active ingredient.

The present invention also provides a cosmetic composition for improving wrinkles containing the polyamine-containing fermented lactic acid bacteria as an active ingredient.

The novel Lactobacillus curvatus -GFC-AJ6 provided from the present invention Spermidine and Spermine by biotransformation through addition of arginine substrate which is the initial precursor of polyamine synthesis pathway to the GFC-AJ6 strain (Accession No .: KCCM11932P) Can be produced. When the composition is applied to a cosmetic base as an effective ingredient, the effect of improving skin whitening or wrinkle is excellent, and therefore, it is excellent as a cosmetic composition.

FIG. 1 is a TLC analysis result of confirming ornithine production ability by culturing lactic acid bacteria isolated from Ganghwa-do turnip kimchi in an MRS medium containing arginine.
FIG. 2 is a photograph showing the results of TLC analysis of polyamine components (Putrescine, Spermidine and Spermine) detected from the fermented product of polyamine-containing lactic acid bacteria of Example 1. FIG.
Fig. 3 shows the results of HPLC quantitative analysis of polyamines from the polyamine-containing lactobacillus fermentation product of Example 1. Fig.
FIG. 4 shows a phylogenetic analysis of the Lactobacillus curvatus GFC - AJ6 strain of Example 1. FIG.
5 is a graph showing the cytotoxicity evaluation of the fermented product of the polyamine-containing lactic acid bacteria of Example 1.
6 is a graph showing the results of DPPH (2,2-Diphenyl-1-picrylhydrazyl) scavenging activity of a polyamine-containing lactic acid fermentation product of Example 1. Fig.
7 is a graph showing the results of intracellular and extracellular melanin activities of a fermented product of a polyamine-containing lactic acid bacterium of Example 1. Fig.
8 is a graph showing the results of promoting Procollagen type I production and increasing expression of COL1A1 gene in the fermented product of polyamine-containing lactic acid bacteria of Example 1.

The present inventors have recently developed a novel strain having the ability to produce polyamines, which is attracting attention as an effective ingredient for cosmetics.

Therefore, in order to develop a novel strain having the ability to produce polyamines, in the present invention, Lactobacillus curvatus GFC - GFC-AJ6 (Accession No .: KCCM11932P) was isolated from turnip kimchi, A lactic acid fermentation product containing Putrescine, Spermidine, and Spermine, which are polyamines produced through bioconversion from arginine, an early precursor of the polyamine synthesis pathway, was prepared using the strain . The inventors of the present invention have found that the antioxidation, skin whitening and wrinkle-reducing effects of the polyamine-containing lactic acid bacteria fermented product can be exerted as an excellent cosmetic composition.

Accordingly, the present invention relates to a Lactobacillus curvatus GFC-AJ6 strain (Accession Number: KCCM11932P) of GRAS (Generally Recognized As Safe) grade having polyamine production ability derived from turnip kimchi .

At this time, the polyamines may be at least one selected from the group consisting of Putrescine, Spermidine and Spermine.

The present invention also relates to a method for producing a polyamine-containing lactic acid bacterium, which comprises culturing a strain of Lactobacillus curvatus GFC-AJ6 (accession number: KCCM11932P) in a medium to which arginine is added Thereby producing a fermented product. In other words, the biotransformation from the Arginine substrate, which is the initial precursor of the polyamine synthesis pathway, by using Lactobacillus curvatus GFC-AJ6 strain (Accession No. KCCM11932P) isolated from the turnip kimchi, To produce Putrescine, Spermidine, and Spermine.

The medium may be an MRS medium containing 0.5 to 1% by weight of arginine. If arginine is added in an amount of less than 0.5% by weight, the content may be insignificant, and polyamine may not be produced in the fermented lactic acid bacteria. If more than 1% by weight of the fermentation product is added, So it is not desirable.

Also, the culturing is performed at 30 to 37 DEG C for 3 to 5 days. If the culture temperature and time are out of the above range, the culturing conditions of the fermenting bacteria are not suitable, and polyamine production and bacterial growth do not occur.

As described above, putrescine, which is a polyamine produced by bioconversion from arginine using a novel Lactobacillus curvatus GFC-AJ6 strain (Accession No .: KCCM11932P) When fermented lactic acid bacteria containing Spermidine and Spermine are produced, skin whitening or wrinkle reducing effect is excellent.

Accordingly, the present invention can provide a skin whitening cosmetic composition containing the polyamine-containing fermented lactic acid bacteria as an active ingredient.

Further, the present invention can provide a cosmetic composition for improving wrinkles containing the polyamine-containing fermented lactic acid bacteria as an active ingredient.

The cosmetic composition for skin whitening or wrinkle improvement according to the present invention can be used as a cosmetic composition for skin whitening or wrinkle improvement which can be used as an external ointment for skin, a cream, a softening longevity, a nutritional lotion, pack, essence, hair tonic, shampoo, rinse, hair conditioner, hair treatment, Skin lotion, milk lotion, nutrition lotion, massage cream, nutritional cream, eye cream, moisturizing cream, hand cream, foundation, nutrition essence, sunscreen, soap, cleansing foam, cleansing lotion, cleansing cream, skin toner, astringent, lotion, milky lotion, But are not limited to, formulations selected from the group consisting of body lotions and body cleansers. The composition of each of these formulations may contain various kinds of bases and additives necessary for formulation of the formulation, and the kinds and amounts of these ingredients can be easily selected by those skilled in the art.

When the formulation of the present invention is a paste, cream or gel, animal fibers, plant fibers, wax, paraffin, starch, tracant, cellulose derivatives, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide are used as carrier components .

When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component. In the case of a spray, in particular, / Propane or dimethyl ether.

In the case of the solution or emulsion of the present invention, a solvent, a solvent or an emulsifier is used as a carrier component, and examples thereof include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, , 3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol or fatty acid esters of sorbitan.

When the formulation of the present invention is a suspension, a carrier such as water, a liquid diluent such as ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, Cellulose, aluminum metahydroxide, bentonite, agar or tracant, etc. may be used.

When the formulation of the present invention is an interfacial active agent-containing cleansing, the carrier component is selected from aliphatic alcohol sulfates, aliphatic alcohol ether sulfates, sulfosuccinic acid monoesters, isethionates, imidazolinium derivatives, methyltaurate, sarcosinates, fatty acid amides Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, linolenic derivatives or ethoxylated glycerol fatty acid esters.

In the cosmetic composition for skin whitening or wrinkle improvement of the present invention, the cosmetic composition for skin whitening or wrinkle improvement may contain 0.01 to 99% of the polyamic acid fermented lactic acid bacteria based on the total weight of the composition, May be contained in an amount of 7.0 to 15%, but is not limited thereto, including an effective amount capable of providing a desired skin whitening or wrinkle-reducing effect.

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not to be construed as limiting the scope of the present invention. It will be self-evident.

Example 1 Isolation and Identification of Polyamine Production Strain

1. Ornithine-producing strain selection

In order to isolate the lactic acid bacteria having ornithine-producing ability from the turnip kimchi samples, firstly, the Kwangju kimchi samples were diluted to an appropriate concentration and then the MRS medium (Proteose peptone No. 3 10 g, Beef extract 10 g, Yeast extract 5.0 g , Dextrose 20g, Polysorbate 80g, Ammonium citrate 2g, Sodium acetate 5g, Magnesium sulfate 0.1g, Manganese sulfate 0.05g, Dipotassium phosphate 2g / l, Difco, USA) And separated into candidate groups in lactic acid bacteria. In order to confirm the ornithine production, the isolated strains were cultured in MRS media supplemented with arsenic substrate at a low concentration of 1.0, 3.0, and 5.0 wt%, and the supernatant was recovered and TLC was used to confirm ornithine production ability. The ornithine production was carried out on Silica gel plate F254 at intervals of 0.5 cm and developed with a solvent of Butanol: Acetic acid:: Water = 5: 2: 2 (v / v / v) Spot and GFC - AJ1 ~ 16 were isolated from ornithine - producing strains. As a result, as shown in Fig. 1, a spot produced by ornithine after incubation of arginine before culturing was confirmed, and it was finally confirmed that ornithine was produced at 1.0 wt%.

2. Preparation of fermented lactic acid bacteria containing polyamines

The arginine-containing MRS liquid medium was prepared by adding arginine substrate to the MRS liquid medium to prepare an arginine-containing MRS liquid medium. After sterilization, the activated candidate strain was inoculated and incubated for 3 days at 37 ° C for activation to produce a polyamine-containing lactic acid fermentation product Respectively. After culturing, the fermented lactic acid bacteria were centrifuged at 14,000 rpm for 15 minutes, and the supernatant was used as an analytical sample.

3. Identification and quantitative analysis of polyamines in fermented lactic acid bacteria

Since the polyamines do not exhibit the extinction, qualitative and quantitative analysis was carried out by dansylation and benzoylation derivatization as a method for identifying the polyamines in the fermented lactic acid bacteria.

5% Perchloric acid was added to 1 ml of the polyamine standard substance and the fermentation product, and the mixture was shaken and centrifuged at 14,000 rpm for 5 minutes. Then, 100 μl of a saturated sodium carbonate solution was added to 50 μl of supernatant. 10 μl of Dansyl chloride solution (10 μg / ml) was added and mixed well. The mixture was allowed to stand in a dark state at 60 ° C for 1 hour. Then, to remove Dansyl chloride, 50 μl of Proline (100 mg / ml) was added to the reaction mixture, and the mixture was allowed to stand at 60 ° C. for 30 minutes. After that, Acetone was removed by a speed vac, 400 μl of Toluene was added thereto and the mixture was centrifuged , The toluene layer was recovered and the solvent was evaporated with a speed vac. The solution was dissolved in 1 ml of methanol and subjected to TLC analysis. Silica gel plate F254 was dispensed at intervals of 0.5 cm and developed with a chloroform: triethylamine = 5: 1 (v / v) solvent to confirm and compare standard spot at 365 nm wavelength to confirm polyamine formation on the TLC . (Fig. 2)

Benzoylation was made alkaline by adding 3 ml of 6M NaOH to 1 ml of the polyamine standard and the supernatant. Benzoyl chloride (5 μl) was added, shaken, allowed to stand for 30 minutes, 1 ml of saturated NaCl and 1 ml of chloroform were added, and the mixture was centrifuged at 3,000 rpm for 10 minutes. The chloroform layer was recovered and the solvent was evaporated in a speed vac. The solution was dissolved in 1 ml of methanol and analyzed by HPLC. Analysis was carried out using a YL9100 HPLC system (Younglin, Korea). Ccolumns were analyzed using a Kinetex C18 (25 cm x 4.6 mm, 5 μm) and a UV detector at 245 nm. The mobile phase was analyzed by injecting 20 μl of the sample under the conditions of Water: Acetonitrile, 60:40 (v / v) at a flow rate of 1.0 mL / min. The polyamine content was calculated from the standard quantitative curve of the peak relative to the concentration of the reference material.

As a result, as shown in FIG. 3, the retention times of putrescine, spermidine and spermine were completely separated into 4.64 minutes, 7.21 minutes, and 11.46 minutes within 12 minutes, respectively. Putrescine, spermidine and spermine were not produced in the other candidate strains, but no spermidine and spermine were produced in the candidate strains. However, the putrescine, spermidine and spermine polyamines were not detected in the fermented lactic acid bacteria using the candidate strain GFC - AJ6 , And contained quantitative results of 23.14 占 퐂 / ml of putrescine, 4.17 占 퐂 / ml of spermidine and 2.13 占 퐂 / ml of spermine. Qualitative and quantitative analysis of three polyamines in the fermented material was completed and it was finally confirmed that the fermented material contained polyamine, which is an effective substance, in the fermented product.

4. Selection strain  16s rRNA  Sequencing

The identification of the polyamine-producing strain GFC-AJ6 was performed by 16S rRNA sequence analysis. (5 'AGA GTT TGA TCM TGG CTC AG 3') and SEQ ID NO: 2: 1492R (5 'TAC GGY TAC CTT GTT ACG ACTT 3') from the gDNA obtained by performing chromosomal DNA extraction PCR was carried out to analyze the nucleotide sequence. Based on the results obtained by performing 16s rRNA sequencing, the homology test was performed through the BLAST search provided by NCBI. As a result of the identification through the nucleotide sequence, the isolate belongs to the genus of Lactobacillus Lactobacillus curvatus was 99.71% similar to Lactobacillus curvatus . Based on the results, Lactobacillus curvatus GFC-AJ6 ( Lactobacillus curvatus GFC-AJ6), deposited at the Korean Microorganism Conservation Center (KCCM) on November 16, 2016, and assigned the deposit number KCCM11932P.

MEGA 6.0 (Megasoftware, USA) program was used to create a schematic diagram by Neighbor-joining method (FIG. 4).

≪ Example 2 > Cytotoxicity test: WST assay

B16-F10 melanoma cells (ATCC ^ⓡ CRL 6475 ™) is a 96 well plate (Corning 3590) in 5 * 10 ³ / well cultured and hFF (Human Foreskin Fibroblast, CCD- 986Sk) (ATCC ⓡ CRL-1947 ™) is 3 * 10 < ³ > / well. After 48 hours, 10% EZ-Cytox (Daiil, 99-EZ3000) was mixed with DMEM media, and the sample-treated media was dispensed at 100 μl per well. After incubation for 2 hours, absorbance was measured at 450 nm wavelength with a microplate reader (Biotek, Synergy HT). Cell viability was calculated as follows in comparison with the untreated group (untreated group).

Cell survival rate (%) = (absorbance of treated group / absorbance of untreated group) * 100

As a result, as shown in FIG. 5, toxicity to B16 / F10 and hFF cells did not show up to the treatment concentration of 4%, and it was confirmed that it could be used as a safe material for cosmetics.

<Example 3> Antioxidant efficacy test: DPPH radical scavenging activity

Free radical scavenging activity of free radical scavenging activity was assessed in order to accelerate aging and to induce immune response by attacking normal cells.

First, fermented lactic acid bacteria containing polyamines were diluted by concentration and distilled water was used as a control. 50 μl of each diluted sample, 200 μl of ethanol, and 250 μl of 0.1 mM DPPH were mixed and reacted at 4 ° C for 30 min in the absence of light, and then absorbance was measured at 520 nm. Ascorbic acid was used as a positive control.

Free radical scavenging (%) = 100- (absorbance of test group addition / control absorbance X 100)

As a result, as shown in FIG. 6, the IC ₅₀ value was found to be 1.46%, and the antioxidative activity of the fermented product of the polyamine-containing lactobacillus was increased depending on the concentration. The antioxidative activity of Ascorbic acid (3 ㎍ / ㎖) which was used as a control group was confirmed to be about 64% at the maximum concentration of 4%.

< Example  7> Whitening efficacy Verification test  : Intracellular · Outer Melanin Analysis

B16-F10 melanoma cells were cultured in ⁶ wells at 1.2 * 10 ⁵ / well. After 24 hours of incubation, α-MSH (Sigma M4135) was treated with 50 nM to induce melanin secretion, and the polyamine-containing lactobacillus fermentate was treated with cells at different concentrations. At this time, phenol-free DMEM media (10% FBS, penicillin (100 unit / ml), streptomycin (100 μg / ml)) was used for melanin analysis. After 48-60 hours, the cells and the culture were transferred to a Conical tube and centrifuged at 3,000 RPM for 10 minutes at room temperature. For the extracellular melanin assay, the cell culture was dispensed into a 96-well plate (corning 3595) at a concentration of 100 μl per well in a total of 4 wells, and the absorbance was measured at 450 nm using a microplate reader.

Cells were collected for analysis of intracellular melanin, washed with 1 ml of DPBS, added with 100 μl of 1N NaOH (10% DMSO), and reacted at 60 ° C for 60 minutes. After completion of the reaction, centrifugation was carried out at 17,000 rpm, and the supernatant was dispensed into 96-well plates in the same manner as extracellular melanin. Absorbance was measured at 450 nm wavelength using a microplate reader. Negative control was not added to α-MSH in both extracellular and intracellular melanin assays. Α-MSH and distilled water were used for control, and β-arbutin was used for positive control.

Intracellular and extracellular melanin contents (%) = (absorbance of treated group / absorbance of control group) * 100

As a result, as shown in FIG. 7, the intracellular melanin contents IC ₅₀ value was 1.29% and the extracellular melanin contents IC ₅₀ value was 1.68%, and the intracellular and extracellular melanin secretion of the polyamin containing lactobacillus- And thus, a whitening effect with excellent melanin formation inhibitory effect was confirmed. At the maximum concentration of 4%, about 73% of the intracellular and extracellular about 71% of the melanin contents inhibition rate were similar to or higher than that of Arbutin (200 ug / mL) used as the control group.

<Example 8> Wrinkle-improving efficacy test: Promotion of procollagen type Ⅰ production and expression of COL1A1 gene

Procollagen type Ⅰ production promoting effect was confirmed by ELISA and COL1A1 gene using real-time PCR. In the ELISA, hFF cells were cultured in 24-wells at 1.2 × 10 ⁴ cells for 24 hours, then treated with serum-free DMEM for 6 hours and treated with serum-free DMEM for 48 hours. After the culture was recovered, Procollagen ELISA (TAKARA MK101) was performed and treated with TGF-β 5 ng / ml as a positive control.

In real-time PCR, hFF cells were cultured in 1.2 × 10 ⁴ cells in 24 wells and treated for 24 hours with serum-free DMEM for 24 hours. Serum-free DMEM was treated with polyamine-containing lactic acid fermentation for 48 hours. RNA was extracted using Nucelospin RNA kit (MN740955) and RNA was quantified using Take3 (Bioteck). cDNA was synthesized using cDNA synthesis kit (Genedepot R5600), and then Real-Time PCR (Biorad) was performed with Sybr green (BR170-8884AP), cDNA and primer. Primer and Real-Time PCR conditions are shown in Table 1 below. Analysis software used CFX manager (Biorad) and β-actin as house keeping gene.

Primer Real Time PCR condition Procollagen SEQ ID NO: 3: F5 '- GACCTCAAGATGTGCCACTC-3'
SEQ ID NO: 4: R 5 '- CCAGTCTCCATGTTGCAGAA-3' Pre-denaturation 95 ° C
Denaturation 94 ° C, 15 s
Annealing 58 ℃, 30s
Extension 72 ° C, 30 s
40 cycles β-actin SEQ ID NO: 5: F5'-CATGAAGTGTGACGTGGACA-3 '
SEQ ID NO: 6: R 5 '- CAGGGCAGTGATCTCCTTCT-3'

As a result, as shown in FIG. 8, the concentration of Procollagen type I was increased to 14% at a maximum concentration of 4% in the ELISA assay. Changes in collagen expression were confirmed by real-time PCR using the COL1A1 gene. The COL1A1 transcription level was increased in a concentration-dependent manner, and 28% COL1A1 expression was further promoted at a maximum concentration of 4% as compared with the TGF-β treatment group, indicating high anti-wrinkle activity.

While the present invention has been particularly shown and described with reference to specific embodiments thereof, those skilled in the art will appreciate that such specific embodiments are merely preferred embodiments and that the scope of the present invention is not limited thereto will be. Accordingly, the actual scope of the present invention will be defined by the appended claims and their equivalents.

Korea Microorganism Conservation Center (KCCM) KCCM11932P 20161116

<110> KANG, Hee-Cheol <120> Novel Lactobacillus curvatus GFC-AJ6, Lactic Acid Bacteria          Fermented Product Comprising Polyamines Using Lactobacillus          Curvatus GFC-AJ6 and Producing Method Thereof <130> P16222 <160> 6 <170> KoPatentin 3.0 <210> 1 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Universal primer 27F <400> 1 agagtttgat cmtggctcag 20 <210> 2 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> Universal primer 1492R <400> 2 tacggytacc ttgttacgac tt 22 <210> 3 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Procollagen primer F <400> 3 gacctcaaga tgtgccactc 20 <210> 4 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Procollagen primer R <400> 4 ccagtctcca tgttgcagaa 20 <210> 5 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Beta-actin primer F <400> 5 catgaagtgt gacgtggaca 20 <210> 6 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> beta actin primer F <400> 6 cagggcagtg atctccttct 20

Claims (9)

Lactobacillus curvatus GFC-AJ6 strain (accession number: KCCM11932P) having three kinds of polyamine producing ability composed of putrescine, spermidine and spermine, .
The Lactobacillus curvatus GFC-AJ6 strain (Accession No. KCCM11932P) according to claim 1, wherein the strain is derived from a turnip kimchi.
delete Lactobacillus curvatus GFC-AJ6 ( Lactobacillus curvatus &lt; RTI ID = 0.0 &gt; GFC-AJ6) &lt; / RTI &gt; having three polyamine production capacities consisting of Putrescine, Spermidine and Spermine in a medium to which arginine is added, AJ6) (Accession No .: KCCM11932P), which comprises culturing a fermented product of lactic acid bacteria containing Putrescine, Spermidine and Spermine.
5. The fermented product of Putrescine, Spermidine and Spermine as claimed in claim 4, wherein the culture medium is an MRS medium containing 0.5 to 1.0% by weight of arginine. Gt;
The method according to claim 4, wherein the culturing is carried out at 30 to 37 ° C for 3 to 5 days. The method for producing a fermented product of lactic acid bacteria containing Putrescine, Spermidine and Spermine.
A fermented product of lactic acid bacteria containing Putrescine, Spermidine and Spermine prepared by the method of any one of claims 4 to 6.
A cosmetic composition for skin whitening comprising the fermented product of lactic acid bacteria containing Putrescine, spermidine and spermine according to claim 7 as an active ingredient.
A cosmetic composition for improving wrinkles comprising the fermented product of lactic acid bacteria containing Putrescine, Spermidine and Spermine according to claim 7 as an active ingredient.

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