Anti-fatigue composition

Abstract

To provide an anti-fatigue composition that eases physical and mental fatigue.SOLUTION: The present inventors have discovered a method for producing an anti-fatigue composition through the combined use of GABA and lactic acid bacteria for easing mental fatigue, and citric acid for easing physical fatigue, thereby completing the present invention.SELECTED DRAWING: Figure 1

Description

Technical Field

[0001] The present invention relates to a composition for anti-fatigue and the like.

Background Art

[0002] The Japanese Society of Fatigue Science defines "fatigue as a state of reduced physical activity ability accompanied by a unique discomfort and a desire for rest caused by excessive physical and mental activities or diseases." When classified by symptoms, it is classified into those caused by physical fatigue and those caused by mental fatigue. Approximately 40% of Japanese people suffer from chronic fatigue, which has become a social problem. Therefore, food components that reduce fatigue have attracted attention. As components that relieve physical fatigue, organic acids such as citric acid and amino acids such as branched-chain amino acids have been reported. As components that relieve mental fatigue, γ-aminobutyric acid (GABA), lactic acid bacteria, etc. have been reported.

[0003] Currently, as a method for producing GABA, which is a component that alleviates mental fatigue, a method of producing GABA by culturing the lactic acid bacterium Levilactobacillus brevis in various media has been reported. For example, a method for preparing a fermented product containing GABA by culturing Levilactobacillus brevis in a GYP medium supplemented with sodium glutamate (Non-Patent Document 1), a method for preparing a fermented product containing GABA by culturing Levilactobacillus brevis NBRC12005 in an MRS medium supplemented with sodium glutamate (Patent Document 1), a method for producing a fermented product containing GABA by culturing Levilactobacillus brevis isolated from Ajinomoto without using a culture medium supplemented with glutamate (Patent Document 2), a method for producing a fermented product containing GABA by adding glutamate to a tomato or celery extract and fermenting it with Levilactobacillus brevis (Patent Document 3), a method for producing a fermented product containing GABA by adding glutamate or its salt to papaya fruit juice and fermenting it with Levilactobacillus brevis (Patent Document 4), and a fermented product containing GABA obtained by statically culturing Levilactobacillus brevis having an anti-allergic effect in a liquid medium (Patent Document 5) have been reported. However, in these reports, the production amount of GBAB is as low as 0.1 mg to 9 g / L, and only the productivity of GABA has been examined. There has been no examination of a composition that combines lactic acid bacteria having effects such as reducing mental fatigue associated with stress in addition to GABA and citric acid that alleviates physical fatigue.

[0004] In addition, a method for producing GABA has been reported in which Lactobacillus brevis, a lactic acid bacterium, is cultured and the obtained cells are reacted with glutamic acid using resting cells (Non-Patent Document 2). However, when pyridoxal phosphate, which is not approved for use in foods in this report, is not used, the production amount of GBAB is 36 g / L. Also, in this method, only the productivity of GABA has been examined, and no attention has been paid to the method for producing a composition that also has lactic acid bacteria having effects such as reducing mental fatigue associated with stress in addition to GABA and citric acid that alleviates physical fatigue. As described above, a composition having components that alleviate physical and mental fatigue is still in the undeveloped stage.

Prior Art Documents

Patent Documents

[0005]

Patent Document 1

Patent Document 2

Patent Document 3

Patent Document 4

Patent Document 5

Non-Patent Documents

[0006]

Non-Patent Document 1

Non-Patent Document 2

Summary of the Invention

Problems to be Solved by the Invention

[0007] An object of the present invention is to provide a composition for anti-fatigue that alleviates physical fatigue and mental fatigue.

Means for Solving the Problems

[0008] The inventors have found a method for producing a composition for anti-fatigue by having both GABA and lactic acid bacteria that alleviate mental fatigue, and citric acid that alleviates physical fatigue, and completed the present invention.

[0009] The present invention relates to the following aspects [1] to [8]. [1] A composition for anti-fatigue containing lactic acid bacteria having GABA-producing ability, and the lactic acid bacteria cell reaction product containing GABA and citric acid. [2] The composition for anti-fatigue according to [1], further containing fruit juice in the lactic acid bacteria cell reaction product. [3] The composition for anti-fatigue according to [1], wherein lactic acid bacteria cultured in a medium containing glutamic acid and / or glutamate are subjected to a cell reaction with a reaction solution containing glutamic acid and / or glutamate, and citric acid and citrate. [4] The composition for anti-fatigue according to [3], further containing fruit juice in the reaction solution. [5] The composition for anti-fatigue according to any one of [1] to [4], wherein the lactic acid bacteria are Levilactobacillus brevis. [6] A method for producing GABA, wherein lactic acid bacteria having GABA-producing ability cultured in a medium containing glutamic acid and / or glutamate are subjected to a cell reaction with a reaction solution containing glutamic acid and / or glutamate, and citric acid and citrate. [7] The method for producing GABA according to [6], wherein the lactic acid bacteria are Levilactobacillus brevis. [8] Levilactobacillus brevis #C-4 having GABA-producing ability.

Effects of the Invention

[0010] In the prior art, there has been no development of an anti-fatigue composition having components that alleviate physical and mental fatigue. However, in the present invention, it is possible to provide an anti-fatigue composition containing a high content of GABA, lactic acid bacteria, and citric acid, for both physical and mental aspects, as well as a method for producing the same.

Brief Description of the Drawings

[0011]

Figure 1

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Modes for Carrying Out the Invention

[0012] The GABA described in the present invention can be produced by reacting lactic acid bacteria cultured in a medium containing glutamate and / or glutamate salt with a reaction solution containing glutamate and / or glutamate salt, as well as citric acid and citrate. Furthermore, lactic acid bacteria having GABA production ability, and a lactic acid bacteria cell reaction product containing GABA and citric acid can be used as an anti-fatigue composition.

[0013] The lactic acid bacteria used in the present invention are not particularly limited as long as they have the ability to produce GABA, but Lactobacillus brevis is preferred. For example, Lactobacillus brevis NBRC12005, Lactobacillus brevis NBRC3345, etc. can be mentioned. In addition, mutant strains derived from the present lactic acid bacteria and having the ability to produce GABA as described above can also be equally used.

[0014] In addition, Lactobacillus brevis #C-4 strain isolated by the present inventors from ume flesh can also be used. The mycological properties of Lactobacillus brevis #C-4 strain isolated by the present inventors from ume flesh are shown below. Incidentally, Lactobacillus brevis #C-4 strain has been deposited with the Patent Microorganisms Depositary, National Institute of Technology and Evaluation (NPMD), an international depositary authority, under the accession number: NITE BP-04015 and is available.

[0015] Lactobacillus brevis #C-4 strain was cultured in MRS agar medium at 30 °C for 48 hours using an anaerobic pouch system. For the grown strains, morphological observation was carried out using an optical microscope, and catalase reaction, oxidase reaction, acid / gas generation from glucose, and oxidation / fermentation of glucose (O / F test) were tested based on the method of Barrow & Feltham et al. The results are shown in Table 1.

[0016]

Table 1

[0017] Next, the fermentation ability with respect to 49 types of carbon sources was confirmed in API 50CHL medium. The results are shown in Table 2. Incidentally, for the fermentation ability, positive is indicated by + and negative is indicated by -.

[0018]

Table 2

[0019] Furthermore, as a result of performing a MALDI microorganism identification test (Biotyper) by MALDI-TOF MS method, the score of Lactobacillus brevis #C-4 strain with Lactobacillus brevis was 2.18. Furthermore, colony PCR was performed using a single colony of Lactobacillus brevis #C-4 strain to amplify approximately 700 bp of the 16s rRNA sequence. Each amplified fragment was cloned into the cloning vector pTA2, and after determining the sequence, bacterial DNA base sequence analysis and molecular phylogenetic analysis (16S rDNA) were performed to identify the strain by BLAST search. As a result, the homology rate of Lactobacillus brevis #C-4 strain with Lactobacillus brevis was 99.8 - 99.9%.

[0020] From the above results of bacterial morphological observation, physiological and biochemical property tests, MALDI microorganism identification test, and bacterial DNA base sequence analysis and molecular phylogenetic analysis, Lactobacillus brevis #C-4 strain is a Gram-positive bacillus that grows under anaerobic conditions, has no motility, does not form spores, has negative catalase reaction and oxidase reaction, ferments glucose to produce gas, and was identified as Lactobacillus brevis based on its fermentation ability for carbon sources.

[0021] The medium for culturing the lactic acid bacteria used in the present invention may be any medium as long as the lactic acid bacteria can grow and achieve the desired GABA production ability or the number of lactic acid bacteria, and any of synthetic media and natural media can be used as long as they contain a carbon source, a nitrogen source, inorganic salts, and other micronutrients required by bacteria.

[0022] Examples of the carbon source include glucose, galactose, fructose, L - arabinose, ribose, etc., and one or more of them can be used. The addition amount of the carbon source is, for example, 0.1 - 30% by weight of the medium, preferably 1.0 - 10% by weight. As the nitrogen source, yeast extract and peptone are preferred, but natural nitrogen sources such as corn steep liquor, soybean protein, milk casein, organic nitrogen sources such as urea, and inorganic nitrogen sources such as ammonium sulfate, sodium nitrate, ammonium nitrate, etc. can also be used in combination. The addition amount of the nitrogen source is, for example, 0.01 - 50% by weight of the medium, preferably 0.1 - 30% by weight, more preferably 0.5 - 10% by weight.

[0023] In addition to these carbon and nitrogen sources, in order to enhance the GABA - producing ability and increase the number of lactic acid bacteria, inorganic salts, oleic acid, etc. can be added. Examples of the inorganic salts include manganese sulfate, manganese yeast, etc., and the addition amount of manganese is preferably 0.001 - 1.0% by weight of the medium. As the oleic acid source, oils and fats containing oleic acid, such as olive oil, safflower oil, sunflower oil, rapeseed oil, etc., and surfactants containing an oleic acid side chain, such as polysorbate 80, polyglyceryl oleate, etc., can be mentioned. The addition amount thereof is, for example, 0.01 - 2.0% by weight of the medium, preferably 0.05 - 1.0% by weight.

[0024] The glutamic acid or its salt used in the present invention is free glutamic acid or a glutamate salt. Examples of glutamate salts include sodium glutamate, potassium glutamate, calcium glutamate, magnesium glutamate, etc. In addition, food materials such as seasonings and protein materials containing free glutamic acid or its salt can also be used. For example, natural seasonings such as kelp extract prepared by extraction from natural raw materials and its powder, vegetable proteins such as soybeans and wheat, and amino acid and peptide mixtures obtained by hydrolyzing animal proteins such as pigs and chickens with acids or enzymes can be used. In the present invention, the higher the concentration of glutamic acid and / or its salt added during cultivation, the higher the expression of glutamate decarboxylase related to the production of GABA in the lactic acid bacteria cell body. Therefore, the addition amount of glutamic acid or its salt, preferably the addition amount of sodium glutamate, is, for example, 0.1 to 50% by weight of the medium, preferably 5.0 to 40% by weight, more preferably 10 to 30% by weight.

[0025] As the culture conditions, it is usually carried out in a liquid medium of the above medium components by shaking culture, static culture, etc. The culture temperature is preferably 15 to 45 °C, more preferably 20 to 35 °C. The initial pH of the medium is preferably 4.0 to 8.0, more preferably 5.0 to 7.0. The culture time is preferably 6 to 72 hours, more preferably 12 to 60 hours.

[0026] By culturing lactic acid bacteria under the above conditions, lactic acid bacteria with high expression of glutamate decarboxylase related to the production of GABA can be obtained. The cells may be collected by centrifugation or used as a culture solution containing the cells.

[0027] In the present invention, a lactic acid bacterium highly expressing glutamate decarboxylase related to the production of GABA is allowed to act on glutamate or glutamate salt, or a substance containing the same, and a resting cell reaction for converting glutamate into GABA is carried out, thereby obtaining a lactic acid bacterium cell reaction product containing GABA, citric acid, and lactic acid bacteria. The reaction solution used in the reaction contains glutamate and / or glutamate salt, and citric acid and citrate. The amount of bacterial cells used in the reaction is preferably 0.1 to 50% by weight, more preferably 0.5 to 40% by weight, and even more preferably 1 to 30% by weight in terms of wet bacterial cells when the reaction solution is 100% by weight.

[0028] The concentration of glutamate or its salt in the reaction solution of the resting cell reaction is preferably 0.1 to 40% by weight, more preferably 1.0 to 30.0% by weight, and even more preferably 5.0 to 25% by weight in terms of the total of glutamate and its salt in the reaction solution.

[0029] The reaction solution for the resting cell reaction is prepared using citric acid and its salt. Examples of citrate include sodium citrate, potassium citrate, etc. The final concentration of citric acid in the reaction solution is preferably 0.05 to 2 M, more preferably 0.1 to 1 M. The pH of the reaction solution prepared with citric acid and its salt is preferably pH 3.0 to 7.0, more preferably pH 4.0 to 5.5. In addition, food materials containing free citric acid or its salt can also be used, and ume juice extracted from ume can be used, for example. For example, if it is ume juice containing 18% citric acid, 0.1 to 20% by weight of ume juice may be contained in the reaction solution, preferably 1.0 to 15% by weight, and a lactic acid bacterium cell reaction product having a mellow fragrance can be obtained by adding ume juice. In addition, in order to enhance the anti-fatigue effect of citric acid, citric acid and its salt can be added after the resting cell reaction.

[0030] The resting cell reaction conditions may be any conditions that do not inactivate the enzyme contained in the raw material. The reaction temperature is preferably 10 to 60°C, more preferably 25 to 50°C, and even more preferably 35 to 48°C. The reaction time is preferably 30 minutes to 60 hours, more preferably 6 hours to 40 hours.

[0031] The lactic acid bacterium having the ability to produce GABA of the present invention, and the lactic acid bacterium cell reaction solution containing GABA and citric acid contain GABA and lactic acid bacteria that relieve mental fatigue, and citric acid that relieves physical fatigue. Therefore, it is optimal as a composition for anti-fatigue. Food and drink containing the anti-fatigue composition can be easily prepared by adding an anti-fatigue composition in the form of powder, liquid, paste, etc. to various foods, and become food and drink having the efficacy of relieving mental fatigue or physical fatigue of GABA, lactic acid bacteria, and citric acid. For example, it can be used in beverages such as tea, coffee, black tea, soy milk, and umeshu, confectionery such as biscuits, candies, and chocolates, side dishes such as croquettes and hamburgers, dairy products such as yogurt and cheese, breads, noodles, etc. In addition, it can be used in health foods such as tablets, capsules, and granules, and can also be used in pharmaceuticals, quasi-drugs, feeds, etc.

[0032] Note that the object of the present invention relates to an anti-fatigue composition composed of a lactic acid bacterium having the ability to produce GABA, a lactic acid bacterium cell reaction product containing GABA and citric acid, which is obtained by reacting a lactic acid bacterium cultured in a medium containing glutamic acid and / or glutamate during culture with a reaction solution containing glutamic acid and / or glutamate, and citric acid and citrate, and a method for producing the same, and does not limit the usage method in any way.

Example

[0033] Hereinafter, the present invention will be specifically described by way of examples, but the technical scope of the present invention is not limited by the following examples. In the present invention, % is all by weight unless otherwise specified.

[0034] [Example 1] (Effect of sodium glutamate concentration added during culture on GABA production amount) 100 mL of Medium containing 2.5% glucose, 2.5% yeast extract, 0.05% manganese yeast, and 0.2% emulsifier was added with sodium glutamate at each concentration (5% (Example 1-1), 10% (Example 1-2), 12.5% (Example 1-3), 15% (Example 1-4), or 20% (Example 1-5)). 10% of the cell suspension of Lactobacillus brevis #C-4 strain isolated from ume flesh was added to this medium, and the culture was carried out for 40 hours under the condition of stirring at 28 °C and 100 rpm. After the culture was completed, wet cells were recovered by centrifugation (10,000 g, 15 minutes). Next, 0.2 g of wet cells, 0.16 g of L-glutamic acid, and 0.48 g of sodium L-glutamate were added to 4 mL of 0.6 M citric acid solution (pH 4.5) prepared with anhydrous citric acid and trisodium citrate, and the resting cell reaction was carried out by shaking at 45 °C for 16 hours. After the reaction, the precipitate was removed by centrifugation (10,000 g, 15 minutes), and the supernatant was recovered to obtain Products 1-1 to 1-5.

[0035] [Evaluation Test 1] 10 μL of 0.1 M boric acid buffer (pH 8.0) was added to each 10 μL of appropriately diluted Products 1-1 to 1-5, mixed with 20 μL of 50 mM 4-fluoro-7-nitrobenzofurazan-ethanol solution, and reacted at 60 °C for 1 minute. After the reaction, 460 μL of 5 mM hydrochloric acid solution was added and mixed, and the GABA content was measured by liquid chromatography. As a result, as shown in Figure 1, the higher the concentration of glutamic acid added during the culture, the higher the GABA content, and the GABA production was good when the sodium glutamate concentration was 12.5% or more.

[0036] [Example 2] (Effect of reaction pH on GABA production in resting cell reaction) To 2 mL of 0.6 M citric acid solution adjusted to various pH values (3.5 (Example 2-1), 4.0 (Example 2-2), 4.5 (Example 2-3), 5.0 (Example 2-4), or 5.5 (Example 2-5)) with anhydrous citric acid and trisodium citrate, 0.1 g of wet cells prepared by fermenting for 48 hours with the medium composition of Example 1-5, 0.08 g of L-glutamic acid, and 0.24 g of sodium L-glutamate were added, so that the initial pH in each reaction solution of Examples 2-1 to 2-5 became 4.5, 4.7, 4.9, 5.3, or 5.5. This reaction solution was shaken at 45 °C for 17 hours to perform a resting cell reaction. After the reaction, the precipitate was removed by centrifugation (10,000 g, 15 minutes), and the supernatant was recovered to obtain Products 2-1 to 2-5.

[0037] Using 10 μL each of appropriately diluted Products 2-1 to 2-5, the GABA content was measured by liquid chromatography using the method described in Evaluation Test 1. As a result, as shown in Figure 2, when the initial pH during the resting cell reaction was 4.9, the GABA production amount was 50 g / L, which was good.

[0038] [Example 3] (Effect of reaction temperature on GABA production amount in resting cell reaction) The resting cell reaction was carried out in the same manner as in Example 2-3, except that the temperature during the resting cell reaction of Example 2-3 was set to 35 °C (Example 3-1), 40 °C (Example 3-2), 45 °C (Example 3-3), or 50 °C (Example 3-4). After the reaction, the precipitate was removed by centrifugation (10,000 g, 15 minutes), and the supernatant was recovered to obtain Products 3-1 to 3-4.

[0039] Using 10 μL each of appropriately diluted Products 3-1 to 3-4, the GABA content was measured by liquid chromatography using the method described in Evaluation Test 1. As a result, as shown in Figure 3, when the temperature during the resting cell reaction was 45 °C, the GABA production amount was 50 g / L, which was good.

[0040] [Example 4] (Effect of reaction time on GABA production amount in resting cell reaction) The resting cell reaction in Example 2-3 was carried out in the same manner as in Example 2-3, except that the reaction time was set to 2 hours (Example 4-1), 6 hours (Example 4-2), 17 hours (Example 4-3), or 24 hours (Example 4-4). After the reaction, the precipitate was removed by centrifugation (10,000 g, 15 minutes), and the supernatant was recovered to obtain Products 4-1 to 4-4.

[0041] Using 10 μL each of appropriately diluted Examples 4-1 to 4-4, the GABA content was measured by liquid chromatography using the method described in Evaluation Test 1. As a result, as shown in Figure 4, the longer the reaction time during the resting cell reaction, the better the GABA production.

[0042] [Example 5] (Effect of the amount of bacterial cells on GABA production in the resting cell reaction) The resting cell reaction in Example 2-3 was carried out in the same manner as in Example 2-3, except that the wet cell mass during the resting cell reaction was set to 0.1 g (Example 5-1), 0.2 g (Example 5-2), or 0.3 g (Example 5-3). After the reaction, the precipitate was removed by centrifugation (10,000 g, 15 minutes), and the supernatant was recovered to obtain Products 5-1 to 5-3.

[0043] Using 10 μL each of appropriately diluted Products 5-1 to 5-3, the GABA content was measured by liquid chromatography using the method described in Evaluation Test 1. As a result, as shown in Figure 5, the more the amount of bacterial cells during the resting cell reaction, the better the GABA production. When 0.3 g of wet bacterial cells were used, the GABA production was 86 g / L.

[0044] [Example 6] (Effect of using ume juice on GABA production in the resting cell reaction) To 1.2 mL of 1 M citric acid solution (pH 4.5) adjusted with anhydrous citric acid and trisodium citrate, plum juice containing 18% citric acid was added so that the plum juice concentration was 0% (comparison), 1% (Example 6-1), 5% (Example 6-2), 10% (Example 6-3) or 15% (Example 6-4), and the total liquid volume was adjusted to 2 mL. To this, 0.1 g of the wet bacterial cells prepared in Example 2-3, 0.08 g of L-glutamic acid and 0.24 g of sodium L-glutamate were added, and the mixture was shaken at 45 °C for 17 hours to carry out a resting cell reaction. After the reaction, the precipitate was removed by centrifugation (10,000 g, 15 minutes), and the supernatant was recovered to obtain a comparative product and products 6-1 to 6-4.

[0045] Using 10 μL each of the appropriately diluted comparative product and products 6-1 to 6-4, the GABA content was measured by liquid chromatography by the method described in Evaluation Test 1. As a result, as shown in Fig. 6, even when plum juice was added, the GABA production amount was good. In product 6-2, GABA was 40 g / L, the number of lactic acid bacteria was 10 billion, and citric acid was 123 g / L, and a composition containing GABA, lactic acid bacteria and plum juice was prepared. In addition, all of the obtained products were imparted with a rich aroma derived from plum juice as compared with the comparative product.

[0046] [Example 7] (GABA productivity when using various Lactobacillus brevis bacterial cells) A resting cell reaction was carried out in the same manner as in Example 1-5 except that Lactobacillus brevis #C-4 strain (Example 7-1) or Lactobacillus brevis NBRC12005 strain (Example 7-2) was used. After the reaction, the precipitate was removed by centrifugation (10,000 g, 15 minutes), and the supernatant was recovered to obtain products 7-1 and 7-2.

[0047] Using 10 μL each of the appropriately diluted supernatant solutions of Examples 7-1 and 7-2, the GABA content was measured by liquid chromatography by the method described in Evaluation Test 1. As a result, as shown in Fig. 7, the GABA production amount was good for both strains of Lactobacillus brevis.

[0048] From the above results, in the present invention, it was possible to simply and inexpensively produce an anti-fatigue composition having effects such as reducing mental fatigue, which has GABA, lactic acid bacteria, and citric acid that alleviates physical fatigue. As shown in Example 1, by adding a high concentration of sodium glutamate during cultivation, culturing, and using the obtained lactic acid bacteria for resting cell reaction, it was found that the productivity of GABA can be increased to about 50 g / L. Also, as shown in Examples 2 to 5, by optimizing the conditions of the resting cell reaction, it was found that the productivity of GABA can be further increased to about 90 g / L. Further, as shown in Example 6, it was found that by adding ume juice to the resting cell reaction, an aromatic anti-fatigue composition containing lactic acid bacteria, GABA, and citric acid can be produced. As shown in Example 7, it was found that GABA can be highly produced by the method of the present invention in various Lactobacillus brevis.

Industrial Applicability

[0049] In the production of GABA using conventional Lactobacillus brevis, the GABA production amount was as low as 0.01 to 9 g / L. In contrast, in the method according to the present invention, by reacting lactic acid bacteria cultured in a medium containing glutamate and / or glutamate salt during cultivation with a reaction solution containing glutamate and / or glutamate salt, and citric acid and citrate, a composition containing high concentrations of GABA, lactic acid bacteria, and citric acid was obtained. As a result, it became possible to provide an anti-fatigue composition containing GABA and lactic acid bacteria having effects such as reducing mental fatigue, and citric acid that alleviates physical fatigue.

Claims

1. An anti-fatigue composition containing lactic acid bacteria having GABA-producing ability, and the lactic acid bacteria cell reaction product containing GABA and citric acid.

2. The anti-fatigue composition according to claim 1, further containing fruit juice in the lactic acid bacteria cell reaction product.

3. The anti-fatigue composition according to claim 1, obtained by subjecting lactic acid bacteria cultured in a medium containing glutamic acid and / or glutamate to a cell reaction with a reaction solution containing glutamic acid and / or glutamate, and citric acid and citrate.

4. The anti-fatigue composition according to claim 3, further containing fruit juice in the reaction solution.

5. The anti-fatigue composition according to any one of claims 1 to 4, wherein the lactic acid bacteria are Levilactobacillus brevis.

6. A method for producing GABA, which comprises subjecting lactic acid bacteria having GABA-producing ability, cultured in a medium containing glutamic acid and / or glutamate, to a cell reaction with a reaction solution containing glutamic acid and / or glutamate, and citric acid and citrate.

7. The method for producing GABA according to claim 6, wherein the lactic acid bacteria are Levilactobacillus brevis.

8. Levilactobacillus brevis #C-4 having GABA-producing ability.

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