KR102014464B1 - Tetragenococcus halophilus WiKim0082, which has betaine-producing ability, and a composition containing the same - Google Patents

Abstract

The present invention relates to a novel Tetragenococcus halophilus WiKim0082 and a composition comprising the same. Tetragenococcus halophilus WiKim0082 according to the present invention is a probiotic having excellent betaine-producing ability, and can be variously used for a purpose of human or animal intestinal treatment, skin moisturizing, or prevention or treatment of dry eye. Furthermore, Tetragenococcus halophilus WiKim0082 can be usefully used as a starter for fermentation and seasoning for flavor enhancement.

Description

Tetrapenococcus halophilus WiKim0082, which has betaine-producing ability, and a composition containing the same}

The present invention relates to a novel tetragenococcus halophilus WiKim0082 and a composition comprising the same.

Tetra gen halophilic microorganisms such as Rhodococcus genus (Tetragenococcus sp.) Is used in a variety of high salt adaptive mechanisms, such as K ⁺ absorption mechanism (K ⁺ uptake system) in order to survive in a high salt environment. For example, strains of the genus Tetragenococcus are known to have osmoprotectants or osmoprotectants such as betaine, choline, proline, ectoine, and glutamate as the salt concentration increases. It uses a mechanism to produce osmotic pressure-controlling substances called compatible solutes or to accept them externally to adapt to high salt environments. Functionality of these substances is known through various studies, among which betaine is known for promoting liver recovery (Patent Document 1), skin moisturizing (Patent Document 2), corneal epithelial cell protective activity (Patent Document 3), etc. It is used in various fields such as food, food and medicine.

On the other hand, it is not yet found that the tetragenococcus halophilus strain among the tetragenococcus genus has a production ability against betaine.

US Patent 5,428,063 Korean Patent Publication No. 2016-0116800 Korean Laid-Open Patent No. 2000-0062425

The present invention is to provide a lactic acid bacteria of the genus tetragenococcus excellent in the ability to produce betaine.

Accordingly, the present inventors have tried to find a probiotic strain having excellent betaine production ability from the traditional fermented food, Tetragenococcus halophilus WiKim0082 ( Tetragenococcus) halophilus WiKim0082) was isolated and identified to complete the present invention.

Tetragenococcus halophilus WiKim0082 ( Tetragenococcus halophilus WiKim0082) according to the present invention is a strain of tetragenococcus halophilus derived from kimchi containing anchovy. Although tetragenococcus halophilus WiKim0082 in the present invention was isolated and identified in kimchi, its access route is not limited thereto.

According to an embodiment of the present invention, 16S rRNA sequencing for identification and classification of microorganisms, which are excellent as separated probiotics and exhibits excellent antimicrobial effect against harmful pathogens, was found to have a nucleic acid sequence of SEQ ID NO: 1. . In addition, the analysis shows that the microorganism is Tetragenococcus It showed the highest molecular phylogenetic relationships and flexible halophilus IAM1676 ^T (FIG. 1).

Therefore, the microorganism of the present invention having the 16S rRNA sequence of SEQ ID NO: 1 was used as the method of tetragenococcus halophilus WiKim0082 halophilus WiKim0082) and was deposited on April 02, 2018 at the National Institute of Agricultural Science, Agricultural Research and Development Institute (Accession Number KACC92226P).

Tetragenococcus halophilus WiKim0082 of the present invention is a gram-positive bacterium and is a facultative anaerobe capable of growing in both aerobic and anaerobic conditions. .

Tetragenococcus halophilus WiKim0082 of the present invention is a probiotic and has a general intestinal effect and an immune enhancing effect of lactic acid bacteria. It is well known that lactic acid bacteria of the genus Lactobacillus have a formal effect immune enhancing effect.

In the present invention, 'probiotics' is understood as a living microorganism that improves the host's intestinal microbial environment in the gastrointestinal tract of animals including humans and has a beneficial effect on the health of the host. Probiotics are living microorganisms with probiotic activity that can have a beneficial effect on the intestinal flora of the host when fed in the form of dried cells or fermented products to humans or animals in the form of single or complex strains.

In the following examples, it was confirmed that the tetragenococcus halophilus WiKim0082 of the present invention shows excellent betaine generating ability. Accordingly, the tetragenococcus halophilus WiKim0082 according to the present invention may be used as a probiotic for various purposes such as the production of betaine as well as the use of human or animal suits, immune enhancing, and the like.

One embodiment of the invention provides a composition comprising Tetragenococcus halophilus WiKim0082, its culture, its lysate, its fermentation or its extract.

Tetragenococcus halophilus WiKim0082 included in the composition according to the present invention may exist as live or dead microorganisms and may also exist in a dried or lyophilized form. Forms and formulation methods of lactic acid bacteria suitable for inclusion in various compositions are well known to those skilled in the art. For example, Tetragenococcus halophilus WiKim0082 is a culture obtained by culturing in a known liquid medium or solid medium, or a fermentation product obtained by culturing the strain and additional components together, or using the strain as an organic solvent. The extract may be formulated in the form of a lysate (or lysate), such as a lysate (or lysate), which is dissolved or crushed or homogenized.

In one embodiment, the composition may be a composition for oral administration comprising the tetragenococcus halophilus WiKim0082 strain present as live or dead bacteria.

In another embodiment, the composition may be a transdermal absorption composition comprising a culture, lysate, fermentation or extract of the tetragenococcus halophilus WiKim0082 strain.

The present invention provides a tetragenococcus halophilus WiKim0082, a culture thereof, a lysate thereof, a fermentation thereof or an extract thereof. The formal formulation according to the present invention can be used for the prevention, treatment, and improvement of gastrointestinal diseases in animals including humans. Preferably, the animals include livestock such as cattle, horses, and pigs. The 'gastrointestinal disease' includes both gastrointestinal harmful bacterial infection and inflammatory bowel disease, for example, infectious diarrhea caused by pathogenic microorganisms (E. coli, Salmonella, Clostridium, etc.), gastrointestinal inflammation, inflammatory bowel disease, neuro enteritis syndrome , Small intestine microbial hypergrowth, enterotropic diarrhea, and the like.

The formal formulation according to the invention is preferably administered orally. The dosage may vary depending on the type of gastrointestinal disease, the severity of the disease, age, sex, race, therapeutic or prophylactic purpose, etc., but in general, 10 to 100 billion animals may be administered per adult.

The present invention provides a composition for enhancing immunity comprising Lactobacillus Corbatus WiKim0082, its culture, its lysate, its fermentation or its extract. It is well known that lactic acid bacteria of the tetragenococcus have a formal effect and an immune enhancing effect.

The present invention also provides a composition for producing betaine comprising tetragenococcus halophilus WiKim0082, its culture, its lysate, its fermentation or its extract.

In the following examples, betaine production ability of the tetragenococcus halophilus WiKim0082 strain was confirmed. Betaine represented by Chemical Formula 1 has trimethylglycine and glycine betaine under different names, and has a quaternary ammonium structure having amphoteric properties in a neutral aqueous solution.

[Formula 1]

Tetragenococcus halophilus WiKim0082 according to the present invention may be included in cosmetics, medicine, nutraceuticals, food, feed, feed additives or lactobacillus starters due to this advantageous action.

Accordingly, the present invention provides a cosmetic composition for moisturizing skin comprising a culture of Tetra Genococcus halophilus WiKim0082, a lysate thereof, a fermented product thereof or an extract thereof as an active ingredient. The present invention also provides a cosmetic composition for moisturizing skin comprising betaine isolated from the tetragenococcus halophilus WiKim0082 as an active ingredient.

The cosmetic composition according to the present invention may further include a well-known moisturizing component in addition to the active ingredient, and may further include an additive commonly used in the art within a range that does not impair the effect of the composition. Such additives include thickeners, viscosity modifiers, pH adjusters, functional ingredients, extracts, antioxidants, stabilizers, fragrances, preservatives, preservatives, vitamins, blockers, dyes, pigments or any other ingredients commonly used in lotion. It may include adjuvants and carriers commonly used in the cosmetic field.

In addition, the formulation of the cosmetic composition according to the present invention may be in the form of a general emulsion formulation and solubilized formulation. For example, lotions such as flexible lotions or nourishing lotions, emulsions such as facial lotions, body lotions, creams such as nourishing creams, moisture creams, eye creams, essences, cosmetic ointments, sprays, gels, packs, sunscreens, makeup bases, liquids Formulations such as foundations such as type, solid type or spray type, powders, cleansing creams, cleansing lotions, makeup removers such as cleansing oils, cleansing foams, soaps, body washes and the like.

In case the formulation of the cosmetic composition of the present invention is a paste, cream or gel, the carrier component is animal oil, vegetable oil, wax, paraffin, starch, tracant, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide. This can be used.

When the formulation of the cosmetic composition of the present invention is a solution or emulsion, solubilizers or emulsions are used as carrier components, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, Dipropylene glycol, butylene glycol, 1,3-butyl glycol oil, glycerol aliphatic ester, PEG-8, polyethylene glycol or sorbitan and the like can be used.

When the formulation of the cosmetic composition of the present invention is a spray, it may further include a propellant such as nitrogen, carbon dioxide gas, chlorofluorohydrocarbon, propane, butane or dimethyl ether.

The cosmetic formulation is a relatively high concentration of Tetra Genococcus halophilus WiKim0082, its culture, its use in the case of wash-off type cosmetics such as makeup remover, detergent, etc., in which the active ingredient stays on the skin within a short period of time. It may include seafood, fermentation thereof or extract thereof. On the other hand, in the case of a leave-on type cosmetic such as a lotion, an emulsion, a cream, an essence, etc., in which the active ingredient stays on the skin for a long time, the concentration of tetragenococcus halophilus is lower than that of the wash-off type cosmetic. WiKim0082, its culture, its lysate, its fermentation or its extract may be included.

The cosmetic composition of the present invention may be used according to a conventional method of use, and the number of times of use may vary depending on the skin condition or taste of the user.

The present invention also provides an eye drop composition comprising a culture of Tetragenococcus halophilus WiKim0082, a lysate thereof, a fermentation product thereof or an extract thereof as an active ingredient.

The eye drop composition may be used for the prevention, amelioration or treatment of dry eye. The term “dry eye” refers to a condition in which the amount of tears is reduced, deficient, or excessively evaporated to cause eye discomfort. This dry eye can cause damage to the ocular surface of the interpalpebral. Symptoms of dry eye may include eye stiffness, foreign body tingling, stinging, or related conjunctival epithelial disorders due to lack of tears in patients with suppressed tear production.

In the present invention, the eye drop composition may further comprise a pharmaceutically acceptable carrier. The pharmaceutically acceptable carrier is commonly used in the manufacture of pharmaceuticals, lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose , Polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, mineral oil, and the like, but is not limited thereto. The composition may also include pharmaceutically acceptable buffers, preservatives (including preservative aids), osmotic modulators, surfactants, solubilizers, stabilizers, emollients, pH adjusters, emollients and / or lubricants. .

In the present invention, the eye drop composition may be provided by filling in a sterile container, it may be provided including an instruction regarding its use. The instructions may be physically attached to a container filled with the eye drop composition or a second container in which the container is packaged, or may be packaged together inside a second container.

The present invention also provides a food composition for producing betaine comprising tetragenococcus halophilus WiKim0082, cultures thereof, lysates thereof, fermented products thereof or extracts thereof as an active ingredient.

When the composition of the present invention is utilized as a food composition, the food composition may include the form of health food or seasonings, beverages, bars and the like. In addition, the food composition containing the strain as an active ingredient may comprise a beverage, such as fermented milk. Accordingly, the present invention provides a fermentation lactobacillus starter consisting of tetragenococcus halophilus WiKim0082, its culture, its lysate, its fermentation product or its extract.

In one embodiment, the food composition may be a seasoning. Tetragenococcus halophilus WiKim0082 strain has excellent betaine production ability involved in umami, and can be used as a seasoning for umami.

The food composition of the present invention may be prepared using a food-compatible and physiologically acceptable adjuvant in addition to the active ingredient, and the adjuvant may include excipients, disintegrants, sweeteners, binders, coatings, swelling agents, lubricants, lubricants. Agents, flavors and the like can be used.

The food composition may be preferably formulated into a pharmaceutical composition including one or more food acceptable carriers in addition to the active ingredient described above for administration.

For example, for formulation in the form of tablets or capsules, the active ingredient may be combined with an oral, nontoxic, pharmaceutically acceptable inert carrier such as ethanol, glycerol, water and the like. In addition, if desired or necessary, suitable binders, lubricants, disintegrants and coloring agents may also be included in the mixture. Suitable binders include, but are not limited to, natural and synthetic gums such as starch, gelatin, glucose or beta-lactose, natural and synthetic gums such as corn sweeteners, acacia, trackercance or sodium oleate, sodium stearate, magnesium stearate, sodium benzo Acetates, sodium acetate, sodium chloride, and the like. Disintegrants include, but are not limited to, starch, methylcellulose, agar, bentonite, xanthan gum and the like. Acceptable pharmaceutical carriers in compositions formulated as liquid solutions are sterile and physiologically compatible, including saline, sterile water, Ringer's solution, buffered saline, albumin injection solutions, dextrose solution, maltodextrin solution, glycerol, ethanol and One or more of these components may be mixed and used, and other conventional additives such as antioxidants, buffers and bacteriostatic agents may be added as necessary. Diluents, dispersants, surfactants, binders and lubricants may also be added in addition to formulate into injectable formulations, pills, capsules, granules or tablets such as aqueous solutions, suspensions, emulsions and the like.

The food composition according to the present invention can be added to various foods. Examples of the food to which the composition of the present invention can be added include beverages, vitamin complexes, and health supplements.

The food composition of the present invention may include ingredients conventionally added in the manufacture of food, and include, for example, proteins, carbohydrates, fats, nutrients, seasonings and flavoring agents. Examples of the above carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose, oligosaccharides and the like; And sugars such as conventional sugars such as polysaccharides such as dextrin, cyclodextrin and the like and xylitol, sorbitol, erythritol. As the flavoring agent, natural flavoring agents (tauumatin, stevia extract (for example, rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be used. For example, when the food composition of the present invention is prepared with a drink and a beverage, citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid, fruit juice, and various plant extracts may be further included.

In addition, the present invention provides a feed additive or feed comprising tetragenococcus halophilus WiKim0082, its culture, its lysate, its fermentation or its extract as an active ingredient.

When used as a feed additive, the composition may be 20 to 90% high concentrate or prepared in powder or granule form. The feed additives include organic acids such as citric acid, fumaric acid, adipic acid, lactic acid and malic acid, phosphates such as sodium phosphate, potassium phosphate, acid pyrophosphate and polyphosphate (polyphosphate), polyphenols, catechins, alpha-tocopherols and rosemary Extracts, vitamin C, green tea extracts, licorice extracts, chitosan, tannic acid, phytic acid, such as any one or more may be further included. When used as a feed, the composition may be formulated in a conventional feed form and may include common feed ingredients.

The feed additives and feeds include cereals such as milled or crushed wheat, oats, barley, corn and rice; Vegetable protein feed, such as feed consisting mainly of rapeseed, soybean, and sunflower; Animal protein feeds such as blood meal, meat meal, bone meal and fish meal; Sugar and dairy products, for example, may further include a dry ingredient consisting of various powdered milk and whey powder, and may further include a nutritional supplement, digestive and absorption enhancers, growth promoters and the like.

The feed additive may be administered alone or in combination with other feed additives in an edible carrier. In addition, the feed additive may be easily administered to the animal as a top dressing or mixed directly in the animal feed or in an oral formulation separate from the feed. When the feed additive is administered separately from the animal feed, it can be prepared in immediate release or sustained release formulation in combination with a pharmaceutically acceptable edible carrier, as is well known in the art. Such edible carriers may be solid or liquid, for example corn starch, lactose, sucrose, soy flakes, peanut oil, olive oil, sesame oil and propylene glycol. If a solid carrier is used, the feed additive may be a tablet, capsule, powder, troche or sugar-containing tablet or top dressing in undispersible form. If a liquid carrier is used, the feed additive may be a gelatin soft capsule, or a formulation of syrup or suspension, emulsion, or solution.

In addition, the feed additives and feed may contain auxiliaries, such as preservatives, stabilizers, wetting agents or emulsifiers, solution accelerators and the like. The feed additive may be used in addition to the feed of the animal by acupuncture, spraying or mixing.

The feed or feed additive of the present invention is applicable to many animal diets, including mammals, poultry and fish.

As the mammal, pigs, cows, sheep, goats, experimental rodents, and experimental rodents, as well as pets (eg, dogs, cats) can be used, and the poultry such as chickens, turkeys, ducks, geese, pheasants, And can also be used for quail, etc., may be used in trout, etc. as the fish, but is not limited thereto.

Feed or feed additives of the present invention can be applied to animal diets can be used for animal growth, immune boosting and the like.

The amount of tetragenococcus halophilus WiKim0082 strain included in the composition according to the present invention may be about 10 ⁶ to 10 ¹² cfu / g based on one time, such as 10 ⁷ to 10 ¹¹ cfu / g, 10 ⁸ to 10 Can be ¹⁰ cfu / g. In the case of administering the strain, it is preferable to administer it in a viable state, and it may be administered before ingestion or attenuated. In addition, in the case of using the culture supernatant, etc., the sterilization process through heat treatment may be additionally performed. The amount of strain and daily intake required to have minimal efficacy may vary depending on the body or health of the ingestor, but may generally be about 10 ⁶ to 10 ¹² cfu / g, such as 10 ⁷ to 10 ¹¹ cfu / g , 10 ⁸ to 10 ¹⁰ cfu / g.

The present invention also provides a method for producing betaine comprising culturing the tetragenococcus halophilus WiKim0082 in a medium containing a salt.

In the betaine production method, the medium for the cultivation of tetragenococcus halophilus WiKim0082 may be 0.1M or more salts (NaCl). In one embodiment, the medium may be one containing the salt at a concentration of 0.1M to 5M, such as 0.2M to 4M, 0.5M to 3M. The step of culturing the tetragenococcus halophilus WiKim0082 may be applied to all methods used in the culturing method of basophilic microorganisms as it is. For example, the step may be a method of culturing the tetragenococcus halophilus WiKim0082 strain in a medium containing at least 0.1M for 12 to 48 hours at a temperature of 25 to 35 ℃, but is not limited thereto.

In the present invention, the betaine production method may further comprise the step of extracting the cultured tetragenococcus halophilus WiKim0082 with an organic solvent. The organic solvent may be used without limitation, conventional extraction solvents such as lower alcohol having 1 to 6 carbon atoms, hexane, acetone, ethyl acetate, chloroform and diethyl ether.

In one embodiment, the organic solvent may be ethanol.

The technique for separating and / or purifying betaine from tetragenococcus halophilus WiKim0082 extract extracted by the organic solvent as described above may be applied to all methods known in the art.

Advantages and features of the present invention and methods for achieving them will be apparent with reference to the embodiments described below in detail. However, the present invention is not limited to the embodiments disclosed below, but may be implemented in various different forms, only the embodiments are to make the disclosure of the present invention complete, and the general knowledge in the technical field to which the present invention belongs. It is provided to fully convey the scope of the invention to those skilled in the art, and the present invention is defined only by the scope of the claims.

Since the tetragenococcus halophilus WiKim0082 according to the present invention exhibits excellent production ability against betaine, it is variously used as a probiotic for the use of human or animal suits, immunity strengthening, prevention and treatment of gastrointestinal diseases and dry eye disease, and skin moisturizing. Can be utilized. Furthermore, it can be usefully used as a starter for fermentation or seasoning for enhancing flavor.

Figure 1 shows the results of systematic analysis of tetragenococcus halophilus WiKim0082 using the 16S rRNA sequence.
Figure 2 shows the results of analyzing the ¹ H-NMR peak of the extract of tetragenococcus halophilus WiKim0082 cells according to the salt (NaCl) concentration.
Figure 3 is a graph confirming the degree of betaine production of tetragenococcus halophilus WiKim0082 according to the salt (NaCl) concentration.

Hereinafter, the present invention will be described in detail through examples. The following examples are merely illustrative of the present invention, and the scope of the present invention is not limited to the following examples.

EXAMPLE

Example 1 Isolation and Identification of Tetragenococcus halophilus Strains

The stock solution of kimchi extract added with anchovy chopped to MRS medium containing 20% NaCl was collected in a loop and cultured in MRS broth. DNA extraction was extracted using the QIAamp DNA Mini Kit (QIAgen, Germany). The extracted DNA was identified by using a 1% agarose gel, PCR was performed using genomic DNA extracted as a template using the primers in Table 1 below to amplify the 16S rRNA gene. Thirty cycles were performed with minutes, annealing 45 ° C. 1 minute, extension 72 ° C. 1 minute 30 seconds. The obtained PCR product was commissioned by Macrogen (Seoul, Korea) to analyze the sequence. Bacterial identification was performed by searching the Ezbiocloud ( https://www.ezbiocloud.net/identify ) site for bacteria with the highest sequencing of the 16S rRNA gene.

Primer (SEQ ID NO) Sequence (5 '-> 3') SEQ ID NO: 2
(Primer_27F) AGAGTTTGATCCTGGCTCAG SEQ ID NO: 3
(Primer_1492R) GGTTACCTTGTTACGACTT

According to the embodiment of the present invention, the strain isolated from the 16S rRNA sequence analysis for the identification of the microorganism, it was found to have a nucleic acid sequence of SEQ ID NO: 1. In addition, as a result of the analysis, the strain was Tetragenococcus It showed the highest molecular phylogenetic relationships and flexible halophilus IAM1676 ^T (FIG. 1).

Thus, the microorganism of the present invention was named Tetragenococcus halophilus WiKim0082 ( Tetragenococcus halophilus WiKim0082), and was deposited on April 02, 2018 at the National Institute of Agricultural Science (KACC) (Accession Number KACC92226P).

Example 2 Confirmation of Betaine Formation of Tetragenococcus halophilus WiKim0082

In order to establish the optimal betaine production conditions of tetragenococcus halophilus WiKim0082 isolated through Example 1, MCl liquid medium was added with different concentrations of NaCl (0.25M, 0.5M, 1M, 2M and 3M). After the addition, the cells were cultured until the OD ₆₀₀ value was 0.8 to adjust the amount of cells under each condition equally to 0.1 g. The cultured strain was recovered and extracted in 80% ethanol and subjected to ¹ H-NMR analysis.

As a result, as shown in Figures 2 and 3, it was confirmed that the production of betaine significantly increased when the concentration of NaCl in the medium is 0.5M or more. In particular, tetragenococcus halophilus WiKim0082 cultured in MRS medium containing 2M NaCl produced about 22mM of betaine, showing the best betaine producing ability among experimental groups.

National Institute of Agricultural Science, Genetic Resources Center KACC92226P 20180402

<110> Korea Food Research Institute <120> Tetragenococcus halophilus WiKim0082, which has betaine-producing          ability, and a composition containing the same <130> P18R10C0554 <160> 3 <170> KoPatentIn 3.0 <210> 1 <211> 1547 <212> DNA <213> Unknown <220> <223> Tetragenococcus halophilus WiKim0082 <400> 1 agagtttgat cctggctcag gacgaacgct ggcggcgtgc ctaatacatg caagtcgaac 60 gctgcttaag aagaaacttc ggttttttct taagtggagt ggcggacggg tgagtaacac 120 gtggggaacc tatccatcag cgggggataa cacttggaaa caggtgctaa taccgcatac 180 ggcttttttt cacctgaaag aaagctcaaa ggcgctttac agcgtcactg atggctggtc 240 ccgcggtgca ttagccagtt ggtgaggtaa cggctcacca aagcaacgat gcatagccga 300 cctgagaggg tgatcggcca cactgggact gagacacggc ccagactcct acgggaggca 360 gcagtaggga atcttcggca atggacgcaa gtctgaccga gcaacgccgc gtgagtgaag 420 aaggttttcg gatcgtaaag ctctgttgtc agcaaagaac aggagaaaga ggaaatgctt 480 tttccatgac ggtagctgac cagaaagcca cggctaacta cgtgccagca gccgcggtaa 540 tacgtaggtg gcaagcgttg tccggattta ttgggcgtaa agcgagcgca ggcggtgatt 600 taagtctgat gtgaaagccc ccagctcaac tggggagggt cattggaaac tggatcactt 660 gagtgcagaa gaggagagtg gaattccatg tgtagcggtg aaatgcgtag atatatggag 720 gaacaccagt ggcgaaggcg gctctctggt ctgtaactga cgctgaggct cgaaagcgtg 780 ggtagcaaac aggattagat accctggtag tccacgccgt aaacgatgag tgctaagtgt 840 tggagggttt ccgcccttca gtgctgcagt taacgcatta agcactccgc ctggggagta 900 cgaccgcaag gttgaaactc aaaggaattg acgggggccc gcacaagcgg tggagcatgt 960 ggtttaattc gaagcaacgc gaagaacctt accaggtctt gacatccttt gaccgcccta 1020 gagatagggt ttccccttcg ggggcaaagt gacaggtggt gcatggttgt cgtcagctcg 1080 tgtcgtgaga tgttgggtta agtcccgtaa cgagcgcaac ccttattgtt agttgccagc 1140 attgagttgg gcactctagc aagactgccg gtgacaaacc ggaggaaggc ggggatgacg 1200 tcaaatcatc atgcccctta tgacctgggc tacacacgtg ctacaatggg aagtacaacg 1260 agcaagccaa gccgcaaggc ctagcgaatc tctgaaagct tctctcagtt cggattgcag 1320 gctgcaactc gcctgcatga agccggaatc gctagtaatc gcggatcagc atgccgcggt 1380 gaatccgttc ccgggccttg tacacaccgc ccgtcacacc acgagagttt gtaacaccca 1440 aagtcggtgc ggcaaccctt aggggagcca gccgcctaag gtgggacgaa tgattggggt 1500 gaagtcgtaa caaggtagcc gtatcggaag gtgcggctgg atcacct 1547 <210> 2 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Primer_27F <400> 2 agagtttgat cctggctcag 20 <210> 3 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> Primer_1492R <400> 3 ggttaccttg ttacgactt 19

Claims (9)

Tetragenococcus halophilus of accession number KACC92226P WiKim0082}. Cosmetic composition for moisturizing skin comprising a culture of Tetragenococcus halophilus WiKim0082 (Accession No. KACC92226P), a lysate thereof, a fermentation product thereof or an extract thereof as an active ingredient. Eye drops for the prevention and treatment of dry eye syndrome comprising a culture of Tetragenococcus halophilus WiKim0082 (Accession No. KACC92226P), a lysate thereof, a fermentation product thereof or an extract thereof as an active ingredient. Tetragenococcus halophilus wikim0082 ( tetragenococcus halophilus WiKim0082; Accession No. KACC92226P), its culture, its lysate, its fermentation or its extract as an active ingredient food composition for producing betaine. The method of claim 4, wherein
The food is a health functional food composition. The method of claim 4, wherein
The food is a seasoning, drink, bar or fermented milk. Tetragenococcus halophilus wikim0082 ( tetragenococcus halophilus WiKim0082; Accession number KACC92226P), its culture, its lysate, its fermentation or its extract as an active ingredient for the production of betaine or feed additives. Betaine production method comprising the step of culturing tetragenococcus halophilus WiKim0082 (Accession Number KACC92226P) in a medium containing a salt. The method of claim 8,
A method of producing betain further comprising the step of isolating and purifying betaine from the cultured tetragenococcus halophilus WiKim0082.

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