KR101739560B1 - Novel Chitinophaga sp. strain having improvement in skin conditions and cosmetic preparation by using same - Google Patents

Abstract

Disclosed are a novel strain having excellent antiseptic functions and excellent antioxidant, whitening and skin wrinkle alleviating efficacy using microorganism separated from soil as a natural cosmetic material, and a cosmetic preparation by using the same. More specifically, provided are a Chitinophagasp sp. NHI-3 (Accession No: KACC92102P) strain separated from soil and a cosmetic preparation by using the same.

Description

[0001] The present invention relates to a chitinopa genus strain having a skin improving effect and a cosmetic preparation using the same. strain taking into consideration skin conditions and cosmetic preparation by using same}

[0001] The present invention relates to a kittenofa genus strain and a cosmetic preparation using the same, and more particularly to a kitenopa genus strain having skin improving effect and a cosmetic preparation using the same.

As interest in skin beauty has increased, various functional cosmetic compositions based on natural materials replacing existing chemical materials have been disclosed.

In the case of preservative functional materials, natural preservatives have been developed due to the harmfulness of existing chemical preservatives, but there are limitations such as production cost, correlation of formulations, side effects due to risks and toxicity.

In the case of whitening functional cosmetics, nine kinds of mulberry extracts are reported as major ingredients, and in Japan, active research on 12 kinds of plant-derived ingredients such as cherry extracts is underway.

The wrinkle-improving functional materials such as mevalonic acid and paeoniflorin have been developed and used. In addition, photolyase (enzyme of healing DNA damage extracted from phytoplankton) ) Has been developed and used as a wrinkle reducing agent.

On the other hand, the functional cosmetics developed so far have mostly visual and temporary improvement effects, but they do not give the continuous and practical effect required by the consumers, and many products use plant-derived natural products as functional cosmetic ingredients, There is a falling problem.

[Prior Patent Literature]

- Open Patent No. 2012-0114540 (released on October 17, 2012)

- Open Patent No. 2012-0107754 (April 4, 2012.)

Accordingly, the present invention is to provide a novel strain having excellent antioxidative, whitening and wrinkle-reducing effects as well as a preservative function by utilizing a microorganism isolated from soil as a natural cosmetic material, and a cosmetic preparation using the same.

In order to solve the above problems, the present invention provides a strain of Chitinophaga sp. NHI-3 (accession number: KACC92102P) isolated from soil.

The present invention also provides a strain of Chitinophaga sp. NHI-3 (accession number: KACC92102P), which comprises 16S rRNA having the nucleotide sequence of SEQ ID NO: 1.

Also provided is a strain of Chitinophaga sp. NHI-3 (accession number: KACC92102P) characterized in that the strain has an antiseptic effect.

Also provided is a strain of Chitinophaga sp. NHI-3 (accession number: KACC92102P) characterized in that the strain further has antioxidant, wrinkle and whitening efficacy.

In order to solve the above-mentioned problems, the present invention provides a cosmetic preparation containing the strain or a culture solution thereof as an active ingredient.

According to the present invention, a cosmetic preparation excellent in antioxidant, whitening and wrinkle-improving effects as well as antiseptic function is provided by using a strain of new strain Chitinophaga sp NHI-3 (accession number: KACC92102P) isolated from soil can do.

1 shows the nucleotide sequence of 16S rRNA gene of NHI-3 ^T strain according to the present invention,
FIG. 2 shows the results of a 16S rRNA gene sequencing analysis of the NHI-3 ^T strain according to the present invention,
3 is a graph showing the results of the antioxidant efficacy analysis in the examples of the present invention,
4 is a graph showing the results of wrinkle-reducing efficacy analysis in the examples of the present invention,
5 is a graph showing the results of the whitening efficacy analysis in the examples of the present invention,
Fig. 6 is a comparative photograph showing the preservative efficacy in the embodiment of the present invention. Fig.

Hereinafter, the present invention will be described in detail with reference to examples. Prior to this, terms and words used in the present specification and claims should not be construed as limited to ordinary or dictionary terms, and the inventor should appropriately interpret the concepts of the terms appropriately The present invention should be construed in accordance with the meaning and concept consistent with the technical idea of the present invention. Accordingly, it is to be understood that the constituent features of the embodiments described herein are merely the most preferred embodiments of the present invention, and are not intended to represent all of the inventive concepts of the present invention, so that various equivalents, And the like.

The inventors of the present invention discovered that a new strain of Chitinophaga sp. Isolated from soil can be used as various functional cosmetic preparations because of its excellent antioxidative, whitening and wrinkle-reducing effects as well as antiseptic function, leading to the present invention .

As a result of searching for microorganisms isolated from the soil, it was confirmed that the microorganisms of the new species had all of the above effects. Based on the nucleotide sequence determination and analysis of the 16S rRNA gene and the DNA-DNA homology test, Was identified as a new microorganism belonging to the genus Chitinophaga sp. And named as " Chitinophaga sp NHI-3 ^T " (hereinafter also referred to as "NHI-3").

According to a preferred embodiment of the present invention, the chitinophaga sp. NHI-3 ^T may include 16S rRNA having the nucleotide sequence shown in SEQ ID NO: 1.

Hereinafter, the present invention will be described in detail with reference to examples.

Example  1: NHI-3 ^T  Isolation and Identification of Strain

(1) Isolation of NHI-3 ^T strain

Soil samples were taken from the forest in summer (Kyonggi University, Korea) and used to remove debris with a 2-mm mesh sieve. 6-polycarbonate transwell plates were used for culturing with various media (Pham & Kim, 2014). 3 g of fresh soil and 3 ml of TSB (Tryptic Soy Broth, 3 ml) / soil suspension (prepared by suspending 1 g of fresh soil in 10 ml of distilled water, 100 μl) ) Inoculum was added. Thereafter, the sample was incubated in a shaking incubator at 28 ° C for 2 weeks. The serial dilutions were plated on the same agar under the same conditions, and the pure colonies were separated after incubation.

(2) Identification of NHI-3 ^T strain

In order to identify the isolated strains, morphological, chemical and genetic characteristics were investigated in the following manner.

Gram stain was determined by Murray et al. (Murray, RGE, Doetsch, RN & Robinow, CF (1994). Determinative and cytological light microscopy, In Methods for General and Molecular Bacteriology , pp. 21-41, Edited by Gerhardt, P., Murray, RGE, Wood, WA & Krieg, NR Washington, DC: American Society for Microbiology. To investigate spore-forming ability, strains were cultured in Schaeffer's medium (0.1% KCl, 0.01% MgCl ₂ , 1.0 mM Ca (NO ₃ ) ₂ , 0.01 mM MnCl ₂ , 0.001 mM FeSO ₄ , and 8 g of nutrient broth per L distilled water) 5 days were incubated at (Kempf, MJ, Chen, F. , Kern, R. & Venkateswaran, K. (2005). Recurrent isolation of hydrogen peroxide-resistant spores of Bacillus pumilus from a spacecraft assembly facility. Astrobiology 5, 391-405.). Cell morphology analysis was carried out 2 days after the culture using an optical microscope (BX50 microscope, Olympus, Japan) at a magnification of 400 ×. Gliding motility was investigated using a hanging drop method. After autoclaving, the pH was adjusted with individual buffers to avoid any change in temperature. The pH was adjusted between 4-11 with the appropriate biological buffer (Breznak, JA & Costilow, RN (1994). Physicochemical factors in growth. In methods for general and molecular bacteriology, pp. 137-154. Edited by P. Gerhardt, R. Murray, WA Wood & NR Krieg . Washington, DC:. American Society for Microbiology), pH 4.0-5.0 , the citrate / sodium phosphate dibasic (citrate / Na ₂ HPO ₄₎ buffer, pH 6.0-7.0, the phosphate (phosphate) buffer, pH 8.0-9.0, the tris ( Sodium bicarbonate / sodium hydroxide (NaHCO ₃ / NaOH) buffer at pH 10.0-11.0. Samples were also grown for 5 days at different growth temperatures of 4, 10, 15, 20, 30, 37, 40, 45, 50 and 55 ° C and at different concentrations of sodium chloride (NaCl) at 0% and 10%. Growth under anaerobic conditions was investigated by culturing the NHI-3 ^T strain on the TSB agar plate in the Oxoid AnaeroGen system under the same conditions as above. Catalase, oxidase, caseinase and amylase activities were determined by 3% (v / v) hydrogen peroxide (H ₂ O ₂ ), 1% (v / v) supplemented with TSB agar ) Bubble formation in tetramethyl-phenylenediamine, 1% (w / v) skimmed milk and 1% (w / v) starch (Smibert & Krieg, 1994 ). Flexirubin pigment was observed by flooding a 20% (w / v) potassium hydroxide (KOH) solution. 0.5% Tween 80 hydrolysis, 0.5% L-tyrosine digestion (Barrow, GI & Feltham, RK A (editors) (1993). Cowan and Steel's manual for the identification of medical bacteria, 3rd edn Cambridge: Cambridge University Press.), 0.4% carboxymethylcellulose (CM-cellulose) (Rautela, GS & Cowling, EB, 1966. Simple cultural test for relative cellulolytic activity of fungi J. Appl. Microbiol 14, 892-898) and Ke Lin School (aesculin) (Swan, A. ( 1954). The use of a bile-aesculin medium and of Maxted's technique of Lancefield grouping in the identification of enterococci (group D streptococci). J. Clin Pathol 7, 160-163.) The test was performed on the TSB agar for 5 days. Flexy Rubin-type pigments were observed in 20% (w / v) potassium hydroxide floated on a colony grown on plates (Reichenbach, H. (1992). The order Cytophagales. In The Prokaryotes. A Handbook on the Edwards by Balows, A., Truper, HG, Dworkin, M., Harder, W., Schleifer, KH New York: Springer. ). API ZYM strips were used to analyze the activity of constitutive enzymes, and API 20 NE and API 50 CH systems (bioMerieux, Marcy I'Etoile, France) were used for carbon sources.

Moore et al. (Moore, E., Arnscheidt. A., Kruger, A., Strompl, C. & Mau, M. (2004). Simulated protocols for the preparation of genomic DNA from bacterial cultures. Molecular Microbiology Ecology Manual , Second Edition DNA extraction was carried out using phenol / chloroform: iso-amylalcohol (1: 1 v / v) in accordance with the method described in Example 1, The rate was settled and removed. DNA purification was then confirmed by electrophoresis. DNA-DNA hybridization was carried out at 2-hour and 30-minute intervals for each measurement using the NHI-3 ^T strain according to the method of Mehlen et al. (Mehlen et al. , 2004) and C. jiangningensis JN53 ^T (Wang et al., 2014) , C. quingshengii JN246 T (Cheng et al., 2015), C. polysaccharea MRP-15 T (Han et al., 2013) and C. niastensis JS16-4 ^T (Weon et al ., 2009). Photobiotin acetate was used as a probe for DNA labeling of NHI-3 ^T strains, and salmon sperm was used as a negative control. In addition, each reference strain was pre-reservedly hybridized as a probe for the NHI-3 ^T strain. The degree of DNA-DNA binding was determined by fluorescence analysis in microplate wells using a counter (1420 Multilabel Counter, Perkin Elmer) and expressed as a percentage. The G + C content was determined using HPLC (Tamaoka, J. & Komagata K. (1984) Determination of DNA base composition by reversed-phase high-performance liquid chromatography, FEMS Microbiol. Lett 25, 125-128; Mesbah, M., Premachandran, U. & Whitman, WB (1989) Precise measurement of the G + C content of deoxyribonucleic acid by high-performance liquid chromatography, Int. J. Syst. Bacteriol 39, 159-167.).

The extracted genomic DNA was used for partial sequencing of the 16S rRNA gene. The 16S rRNA was subsequently amplified using a universal bacterial primer set 27F and 1492R, Pham, VHT & Kim, J. (2014). Bacillus thaonhiensis sp. Nov. , A new species, isolated from the forest soil of Kyonggi university using a modified culture method, Curr. Microbiol 68, 88-95. (PRISM BigDye Terminator v3.1 cycle sequencing kit, Applied Biosystems, Foster City, CA, USA) was used for purification of the PCR product, and a multiscreen-filter plate (Millipore Corp., Bedford, MA, USA) Sequencing was performed using primer 518F (5 '(CCA GCA GCC GCG GTA ATA CG) 3') and primer 800R (5 '(TAC CAG GGT ATC TAA TCC) 3' . The above procedure was carried out at 95 ° C for 5 minutes, followed by cooling for 5 minutes and analyzed using a DNA analyzer (ABI Prism 3730XL DNA analyzer, Applied Biosystems, Foster City, CA, USA). Finally, the entire sequence of the nearly completed 16S rRNA gene was assembled using the program (SeqMan software, DNASTAR Inc., Madison, WI, USA). To generate the phylogenetic tree, the relevant FASTA sequence was obtained from the GenBank database. MEGA6: Molecular Evolutionary Genetics Analysis Version (MEGA6.06) was used for sequencing and phylogenetic reconstruction (Tamura, K., Stecher, G., Peterson, D., Filipski, A. and Kumar S. Molol Biol Evol 30: 2725-2729.). The optimal model was confirmed based on the minimum Bayesian information criterion value (Nei & Kumar, 2000). A Tamura 2-parameter model with uniformity was used as an optimal means for neighbors-joining (NJ) analysis. The reliability of the phylogenetic tree was estimated to be 1,500 repetitions of bootstrap values (Felsenstein, J. (1985).) Confidence limit on phylogenies: an approach using the bootstrap Evolution 39, 783-791. Fig. 2 shows the phylogeny of the 16S rRNA gene sequence of the NHI-3 ^T strain.

The antibiotic resistance was evaluated as an antibiotic-containing disc applied to the TSB agar plated with 100 μl of the bacterial culture for 5 days. The antibiotic discs contained 10 μg of ampicillin, 30 μg of chloramphenicol, 10 μg of gentamicin, 30 μg of kanamycin, 30 μg of nalidixic acid, 30 μg of noviocin 5 μg of rifampicin, 10 μg of penicillin G, 10 μg of streptomycin, 30 μg of tetracycline, 23.75 μg of sulfamethoxazole / 1.25 μg of trimethoprim, And 30 μg of neomycin (Lin et al ., 2009). After 2 days of incubation, the growth inhibition area around each disk was measured and evaluated as susceptible when the characteristics of the strains were more than 11 mm, partial resistance when the strains were 8 to 10 mm and resistant when the strains were less than 8 mm (Nokhal, TH & Schlegel, HG (1983) Taxonomic study of Paracoccus denitrijicans . Int J Syst Bacteriol 33, 26-37.).

Fatty acid profiles were obtained according to the Microbial Identification System (MIDI, Inc., Newark, DE, USA) (Sasser, M. (1990) Identification of bacteria by gas chromatography of fatty acids. 101. Newark, DE: MIDI Inc.). Cells cultured on TSBA (Trypic Soy Broth Agar) for 2 days at 28 ° C in exponential growth phase for fatty acid extraction were identified in the third section of agar plate. Saponification, methylation, extraction and identification were performed to complete the process.

After growth in TSB agar, lyophilized cells were used for isoprenoid quinone and polar lipid extraction. The cells were heated to 60-80 ° C in 2 ml of a methanol: water (100: 10, v / v) mixture containing 0.3% NaCl and the petunum ether was added to dissolve and evaporate the quinone Acetone (Minnikin, DE, O'Donnell, AG, Goodfellow, M., Alderson, G., Athoy, M., Schaal, A. & Parlett, JH bacterial isoprenoid quinones and polar lipids. J. Microbiol. Methods 2, 233-241.). The purified quinone component was analyzed by HPLC (Hairaishi et al ., 1996). Polar lipids were extracted using a mixture of chloroform: methanol (2: 1) and analyzed by two-dimensional thin-layer chromatography (TLC) using other solvents (Komataga & Suzuki, 1987; Minnikin et al ., 1984). Spots with a range of colors and positions for each type of polar lipid were identified with the appropriate reagents as follows. Ethanolic molybdophosphoric acid (20 wt% solution in ethanol; Sigma-Aldrich, Germany) was used for the total lipid profile, ninhydrin for causative lipid containing amino groups, The Zinzadze reagent was used for the fat containing glycerol fat and the Dragendorff reagent was used for the choline, which is represented by α-naphtholsulfuric acid.

The NHI-3 ^T strain grows very broadly at 15 to 40 ° C, preferably at 28 to 30 ° C, using TSB (optimum pH 7 to 8, maximum 5.5% sodium chloride concentration). Colony size is 1 ~ 2mm in diameter after incubation for 5 days and is orange, annular, mucinous and convex. The cell size is approximately 1.8 to 2.7 mu m in length and 0.7 to 0.84 mu m in width. Catalase, oxidase activity and Flexirubin-type pigment are positive. Other morphological characteristics are shown in Table 1 below.

The NHI-3 ^T strain shows strong resistance to novobiocin, ampicillin and kanamycin and is resistant to rifampicin, chloramphenicol, tetracycline, sulfamethoxazole, ) / Trimethoprim, nalidixic acid, gentamicin, penicillin G and streptomycin.

As with all other microorganisms known as Chitinophaga , MK-7 was found to be a major quinone (Kampfer, P., Young, CC, Sridhar, KR, Arun, AB, Lai, (2006), Transfer of [ Flexibacter ] sancti , [ Flexibacter] filiformis , [ Flexibacter ] japonensis , and [ Cytophaga ] arvensicola to the genus Chitinophaga and description of Chinitophaga skermanii sp . Microbiol 56, 2223-2228.). The polar lipid component of the NHI-3 ^T strain was identified as phosphatidylethanolamine (PE), unphosphorous aminophospholipid (APL) and uncharged polar lipid as reported in other Chitinophaga species (Ul, S., Lee, H., Whang, S. (2014) Chitinophaga polysaccharide sp. nov., an Exopolysaccharide-producing Bacterium Isolated from the Rhizoplane of Dioscorea japonica . J. Syst. Evol., Microbiol 64, 55-59.). The major components identified were C _{15: 0} and iso C _{16: 0} w5c. The fatty acid composition of this strain is similar to that of the strains associated with the genus Chitinophaga but has some distinct quantitative differences from other reference strains. Especially, C _{15: 0} showed higher contents than other strains. The minor components (≤10%) were summed feature 4 (C _{17: 1} anteiso B / iso I), C _{16: 1} w11c and C _{16: 0} .

The G + C molar content of the genomic DNA of the NHI-3 ^T strain is 48.2%, similar to other related species (see Table 1). The NHI-3 ^T strain showed a DNA-DNA relatedness of 55% ( C. jiangningensis JN53 ^T ), 49% ( C. quingshengii JN246 ^T ) 45% ( C. polysaccharea ) and 40% ( C. niastensis JS16-4 ^T ). This similar figure is between 25% and 70%, indicating that this species is the same species as the reference strain and is a different species. The strains with the highest 16S rRNA similarity to NHI-3 ^T strain are as follows. C. jiangningensis JN53 ^T (97.01%), C. quingshengii JN246 ^T (96.42%), C. polysaccharea MRP-15 ^T (96.12%) and C. niastensis JS16-4 ^T (96.06%). From the phylogenetic tree, it is clear that the NHI-3 ^T strain belongs to the Chitinophaga strain (see FIG. 2).

From the results of morphological, chemical and genotypic polyphase analysis, it can be known that NHI-3 ^T is a new species in Chitinophaga , named as Chitinophaga sp. NHI-3 ^T , As of November 04, KACC92102P was granted accession number from National Academy of Agricultural Science.

Kitty worm Chitinophaga sp) NHI-3 ^T Taxonomic description of

The NHI-3 ^T strain is gram-negative, is strictly aerobic, and does not form spores. Optimal growth conditions were pH 7 ~ 8, sodium chloride (NaCl) concentration of 0 ~ 2.0% and 32 ~ 37 ℃ in TSB and NB medium. The allowable temperature is maximum 40 ℃ and minimum 15 ℃. This strain forms flexirubin-like pigments. No casein, starch, DNA, tyrosine or Tween 80 is hydrolyzed. Catalase and oxidase activity are positive. Alkaline phosphatase, leucine arylamidase, acid phosphatase, naphthol-AS-BI-phosphohydrolase, and N (N-acetyl-β-glucosaminidase). Trypsin is weakly produced, and esterase (C4), esterase lipase (C8), lipase (C14),? -Chymotrypsin,? -Galactosidase galactosidase, valine arylamidase, cysteine arylamidase,? -galactosidase,? -glucuronidase,? -glucuronidase,? -glucuronidase, -Glucosidase,? -Glucosidase,? -Mannnosidase, and? -Fucosidase in the absence of the enzyme. Nitrate is not reduced to nitrite or nitrogen gas. D-glucose (D-glucose) is weakly fermented. Urease and 4-nitrophenyl-β-D-galactopyranoside, but not L-arginine and L-tryptophan (L- triptophane. It hydrolyzes esculin ferric citrate but does not hydrolyze gelatin. D-glucose, D-mannose, N-acetyl-glucosamine and D-maltose, but L-arabinose (L -arabinose, D-mannitol, potassium gluconate, capric acid, adipic acid, and trisodium citrate. MK-7 is the major menaquinone. The major polar lipid components are phosphatidylethanolamine (PE), a large amount of unmodified aminophospholipid (APL) (1-5) and a small amount of unmodified polar lipid (UL). The major fatty acid composition is C15 _{: 0} , iso-C16 _{: 0} w5c. The DNA G + C content is 48.2%. The type strain NHI-3 ^T was isolated from the rooted-soil of the forest (Kyonggi University, Suwon, South Korea).

Example  2: NHI-3 ^T  Effectiveness of the strain

For efficacy test of a cosmetic use of a NHI-3 ^T strain NHI-3 ^T strain antioxidant, wrinkle as follows by using a (indicated by 'V6') Bacillus sp as comparative strains and (indicated by 'V38') , Whitening, and preservative efficacy.

(1) antioxidant efficacy

The DPPH radical scavenging activity of the 1-butanol extract of each strain was measured according to the following experimental conditions (see Equation 1 below) and compared with the standard vitamin C: Are shown in Table 3 and FIG.

[Experimental Conditions]

- 0.1 mM of DPPH (1,1-Diphenyl-2-picrylhydrazyl)

- Methanol

- A range of concentrations of samples and standard Ascorbic acid (10-45 μg / ml, with 0.5 interval)

(2) Wrinkle-improving efficacy

Elastase activity of the 1-butanol extract of each strain was measured under the following experimental conditions (see Equation 2 below) and compared with oleic acid, a standard substance, , And the results are shown in Table 5 and FIG.

[Experimental Conditions]

- 0.2 M Tris-HCl (pH 8.0) (Buffer)

- 0.5 mM N-Suc- (Ala) 3-nitroanilide (Substrate)

- Porcine pancreatic elastase (PPE) (3.5 U / ml of 0.2 M Tris-HCl) (Enzyme)

(3) Whitening efficacy

Tyrosinase activity was measured for the 1-butanol extract of each strain under the following experimental conditions (see Equation 3 below) and compared with arbutin, a standard substance, Are shown in Table 7 and FIG.

[Experimental Conditions]

Phosphate buffer (100 mM, pH 6.8)

- 1.5 ml Mushroom tyrosinase (3000 U / ml in 50 mM Phosphate buffer)

- 3 mM L-tyrosine (Substrate)

- Concentration of samples and standard Arbutin ranging from 10-45 / / ㎖, with 0.5 interval)

(4) Preservative efficacy

The 1-butanol extract of each strain was tested for inoculation and antibacterial activity under the following experimental conditions and the results are shown in Table 8, Cfu value over time).

[Experimental Conditions]

- Antibiotic compounds: 20 mg / ml

- Emulsion: 9 mL, autoclaved

- Pathogen cell: 10 ⁶ cells / ml

>>>> In total volume of 10 ml

Referring to the above experimental results, it can be confirmed that the NHI-3 ^T strain according to the present invention has an antioxidative effect to some extent, and has excellent wrinkle-improving, whitening and antiseptic properties.

While the preferred embodiments of the present invention have been disclosed for illustrative purposes, those skilled in the art will appreciate that various modifications, additions and substitutions are possible, Such modifications and changes are to be considered as falling within the scope of the following claims.

National Academy of Agricultural Sciences KACC92102P 20151104

<110> INNOGENE CO., LTD. <120> Novel Chitinophaga sp. strain having improvement in skin          conditions and cosmetic preparation by using same <130> NP15-11111 <160> 1 <170> Kopatentin 2.0 <210> 1 <211> 1371 <212> DNA <213> Chitinophaga sp NHI-3T <400> 1 atgcagtcga ggggcagcgc aggtagcaat accgggcggc gaccggcaaa cgggtgcgga 60 acacgtacgc aaccttcctt caagcgggga atagcccaga gaaatttgga ttaatacccc 120 ataatactat tgagaggcat cttttggtag ttaaagattt atcacttgaa gatgggcgtg 180 cgtctgatta ggtagttggt gaggtaacgg ctcaccaagc cgacgatcag taactggcgt 240 gagagcgcga ccagtcacac gggcactgag acacgggccc gactcctacg ggaggcagca 300 gtaaggaata ttggtcaatg gacgcaagtc tgaaccagcc atgccgcgtg gaggatgaag 360 gccctctggg ttgtaaactt cttttatctg ggacgaaaca cttcttttct aaggagcttg 420 acggtaccag aggaataagc accggctaac tccgtgccag cagccgcggt aatacggagg 480 gtgcaagcgt tatccggatt cactgggttt aaagggtgcg taggcggact tgtaagtccg 540 tggtgaaatc tccaggctta acctggaaac tgccatggat actataagtc ttgaatgttg 600 tggaggttag cggaatagtt catgtagcgg tgaaatgctt agatatgacc tagaacacca 660 attgcgaagg cagctggcta cacaataatt gacgctgagg cacgaaagcg tggggatcaa 720 acaggattag ataccctggt agtccacgcc ctaaacgatg attactcgac atttgcgata 780 tattgtaagt gtctgagcga aagcattaag taatccacct gggaagtacg accgcaaggt 840 tgaaactcaa aggaattgac gggggtccgc acaagcggtg gagcatgtgg tttaattcga 900 tgatacgcga ggaaccttac ctgggctaga atgcagattg accgtgggtg aaagctcatt 960 ttgtagcaat acacagtctg taaggtgctg catggctgtc gtcagctcgt gccgtgaggt 1020 gttgggttaa gtcccgcaac gagcgcaacc cctatcacta gttgccagca cttcgggtgg 1080 gaactctagt gaaactgccg tcgtaagacg cgaggaagga ggggatgatg tcaagtcatc 1140 atggccttta tgcccagggc tacacacgtg ctacaatggt agagacaaag ggctgctact 1200 tggtaacaag ctgctaatct caaaaactct atctcagttc ggattgaggt ctgcaactcg 1260 acctcatgaa gctggaatcg ctagtaatcg tatatcagca atgatacggt gaatacgttc 1320 ccggaccttg tacacaccgc ccgtcaagcc atgaaagccc ggggggacct g 1371

Claims (5)

Chitinophaga sp NHI-3 (accession number: KACC92102P) strain isolated from soil. The method according to claim 1,
Wherein said strain comprises 16S rRNA having the nucleotide sequence shown in SEQ ID NO: 1 (Accession No .: KACC92102P). The method according to claim 1,
Wherein said strain has an antimicrobial activity against at least one bacterium selected from the group consisting of Pseudomonas aeruginosa, Staphylococcus aureus, Escherichia coli, Bacillus subtilis and Candida albicans (accession number: Chitinophaga sp) NHI-3 KACC92102P) strain.
The method of claim 3,
Wherein said strain further has antioxidant, wrinkle-improving and whitening efficacy; and Chitinophaga sp NHI-3 (Accession No .: KACC92102P) strain. A cosmetic preparation containing the strain of any one of claims 1 to 4 or a culture thereof as an active ingredient.

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