KR101106668B1 - The preparing method for natural GABA using serial fermentation of plant originated lactic acid bacteria with GABA-producing ability and protease-producing ability - Google Patents

Abstract

The present invention relates to a method for producing a gama (gamma-aminobutyric acid, GABA, gamma aminobutyric acid) -containing fermentation, and a food or beverage comprising the same.

As described above, the present invention is characterized in that natural GABA-containing fermented products are prepared using two-stage fermentation of vegetable lactic acid bacteria having protease generating ability and Gaba generating ability.

According to the present invention, fermented products having a naturally increased Gaba content are prepared using lactic acid bacteria having protease activity and vegetable lactic acid bacteria having glutamate decarboxylase activity, using various materials including glutamic acid-containing proteins as main ingredients. can do.

GABA, glutamic acid, protease lactic acid bacteria, vegetable lactic acid bacteria

Description

The preparing method for natural GABA using serial fermentation of plant originated lactic acid bacteria with GABA-producing ability and protease-producing ability}

The present invention relates to a method for producing a gama (gamma-aminobutyric acid, GABA, gamma aminobutyric acid) -containing fermentation, and a food or beverage comprising the same. More specifically, the present invention is a natural fermentation source containing a large amount of glutamic acid, such as tomato and rice bran as the main fermentation source, and then converted into glutamic acid using a lactic acid bacterium having protease activity, By sequentially culturing the lactic acid bacteria convert the converted glutamic acid to gamma-aminobutyric acid (GABA), and relates to a method for producing tomato fermented products or rice bran fermented products containing natural Gaba without the addition of glutamic acid from the outside.

The fermented product thus produced contains various bioactive substances and organic acids by fermenting lactic acid bacteria as well as Gaba.

Gaba is a non-protein constituent amino acid, which is not only a neurotransmitter in the brain, but also a variety of functions such as brain function, mental stability, blood pressure lowering, diuresis, liver function improvement, obesity prevention, alcohol metabolism and deodorization. Has physiological function In particular, it is registered as a medicine and is used for the treatment of headache, tinnitus and decreased motivation caused by stroke or cerebral artery sequelae.

GABA is known to be widely distributed in various vegetables, fruits, rice and soybeans, but its low content does not show physiological effects. Recently, in Japan, the GABA-rich green tea, rice germ, germinated brown rice, etc. are sold to the food industry and consumers. Interest is rising.

According to a recent study, a study in which tea leaves were produced by anaerobic treatment of tea leaves with N ₂ gas for 6 to 8 hours to drink high-GABA-containing teas in patients with high blood pressure, not only lowered blood pressure but also did not accumulate in the body. It has been proven that it can be used as a natural health drink with no side effects. In addition, the results of physiological function and application experiments of rice germ and germinated brown rice product with high GABA content showed that it increases brain blood flow to increase oxygen supply and activates brain cell metabolism to prevent autonomic ataxia, stroke and dementia. It is effective in promoting and insomnia, blood pressure lowering effect is excellent in circulatory disease improvement effect, liver function and alcohol metabolism promoting function has also been found to have a hangover removal function. As such, GABA is attracting attention as a material having excellent physiological activity in recent years, and thus, it is a functional material that can secure infinite marketability depending on its application.

In the present invention, a technology for producing GABA having the above-described superiority as a natural substance by two-stage fermentation of materials such as tomatoes using lactic acid bacteria.

Lactic acid bacteria (LAB) is an industrially important microorganism that is widely applied worldwide for the production of various fermented food products, including fermented dairy products such as cheese, fermented milk and lactic acid bacteria beverages. In particular, most of the fermented milk marketed in Korea can be said to be a representative food prepared to easily consume live lactic acid bacteria.

Lactic acid bacteria ingested through fermented milk promote the growth of intestinal useful bacteria, while improving the balance of the intestinal flora so that beneficial bacteria take the lead by inhibiting the growth of harmful bacteria, resulting in ① prevention of intestinal infections and food poisoning, ② intestines It has been found to have various physiological functions, including anti-corruption, ③ prevention of constipation by promoting bowel movement, ④ increased immunity, ⑤ inhibition of carcinogenesis, and ⑥ production of vitamin B group. Therefore, in the West, such as Europe, yogurt, lactic acid bacteria beverages and probiotic products in the form of powders, granules and tablets form the largest market as functional foods.

Recently, apart from the lactic acid bacteria isolated from milk and cheese, the advantages of vegetable lactic acid bacteria have been highlighted. Plant Lactobacillus is called POLAB (Plant Originated Lactic Acid Bacteria) and is treated differently from general lactic acid bacteria and refers to lactic acid bacteria isolated from fermented foods of plant raw materials such as pickles and kimchi that have been ingested in the Orient. Vegetable lactic acid bacteria are more than 10 times more varieties than general animal lactic acid bacteria and have excellent adaptability to the external environment. That is, it is a strain that can grow in an environment with high salt concentration and low pH, such as pickles, or in an environment with many antibacterial substances such as garlic, such as kimchi. Kimchi in Korea has high salt content, low pH, and high content of natural antibacterial substances. Therefore, various kinds of bacteria isolated from kimchi are representative vegetable lactic acid bacteria, and thus, mature kimchi is a representative report of vegetable lactic acid bacteria (POLAB).

In the case of ingesting vegetable lactic acid bacteria having such characteristics, unlike animal lactic acid bacteria, resistance to gastric acid is high, the survival rate in the stomach is high, and the intestine survives in many cases. In addition, because they grow in a harsh environment, the productivity of various bioactive substances is often excellent. Representative materials include antimicrobial peptides, immune enhancing substances and GABA.

Lactic acid bacteria are generally not microorganisms used to produce enzymes, and most enzymes are produced using Bacillus or mold. In particular, strains showing strong protease activity are known as Bacillus sp. Isolated from Doenjang and Cheonggukjang. However, there is a problem in the production of fermented foods using protease produced during fermentation of Bacillus strains. First of all, the fragrance component produced by the breakdown of protein by protease is very unique and difficult to apply to food. Of course, in the case of Doenjang or Cheonggukjang, it is not a big problem as a product ingested by using the inherent smell, but recently, research is being conducted to apply strains that generate less smell for generalization of Cheonggukjang. However, in the case of Cheonggukjang, which has a low odor, most of the processes reduce the action of the produced protease by reducing the fermentation time. Therefore, fermented products using Bacillus have not yet completely solve the problem that occurs in the fragrance.

In addition, most of the products related to the intake of live bacteria of domestic consumers focus on lactic acid bacteria. This is because the first probiotic product to drink in Korea has settled in the market based on yogurt. In fact, the live bacteria that are consumed through doenjang, cheonggukjang, and even kimchi are not widely recognized.

Therefore, the development of a functional product using lactic acid bacteria that produce protease will be recognized as a value domestically and globally. The present inventors have already secured the plant lactic acid bacteria that convert glutamic acid to Gabba through the previous research process. In addition, as a result of searching for domestic salted fish, lactic acid bacteria showing protease activity could be obtained.

There are many patents related to GABA, but most of them are methods for increasing the GABA content of tea by drying methods (Registration Nos. 10-0235110 and 10-0367055) and patents relating to various efficacy of GABA (Application Nos. 10-2001-0009589 and 10- 2000-0086507), and patents related to the production of Gabba use a single lactic acid bacterium, such as controlling germination conditions to increase the Gabb content of brown rice (Application Nos. 10-2003-0020882 and 10-2001-0037685). It is limited to the method of converting externally added glutamic acid to Gabba (application number 10-2002-0027264). Therefore, it can be said that there are many differences in terms of configuration and function from those intended to be pursued in the present invention, which naturally produce Gabba using lactic acid bacteria having two functions.

Although somewhat implied by the above, it is an object of the present invention to provide a method for producing a fermentation product comprising Gabba using a plant lactic acid bacterium having protease producing ability and Gaba producing ability.

Another object of the present invention is to provide a method for producing a fermentation product comprising natural gaba without addition of glutamic acid from the outside.

Still another object of the present invention is to provide a food or beverage comprising a Gaba-containing fermented product prepared according to the above production method.

The present inventors made diligent research efforts to build a natural gaba production system using plant lactic acid bacteria having protease generating ability and gaba generating ability. As a result, this invention was completed by confirming the manufacturing method of the fermentation product which naturally contains a Gaba component using the raw material containing glutamic acid as a protein component as a main component.

The present invention for achieving the above object is basically characterized in that the production of natural GABA (GABA) -containing fermentation using two-stage fermentation of vegetable lactic acid bacteria having protease and ability to produce Gaba.

Specifically, the present invention is characterized by including the following steps.

(a) preparing a medium comprising a raw material containing glutamic acid;

(b) inoculating the medium with lactic acid bacteria having protease-producing ability;

(c) culturing the lactic acid bacteria to produce a fermentation product containing glutamic acid;

(d) inoculating the fermented product with the vegetable lactic acid bacteria having the ability to produce Gabba; And

(e) culturing the lactic acid bacteria to produce a fermentation product containing gabba.

At this time, the raw material containing glutamic acid in the production step (a) of the medium is characterized in that the raw material containing a protein containing glutamic acid as a component. More specifically, the material containing a protein comprising glutamic acid is characterized in that the tomato or rice bran.

In addition, the lactic acid bacteria inoculated in step (b) is characterized in that the strain of the genus Lactobacillus . This strain is preferably a Lactobacillus para casei (Lactobacillus paracasei), Lactobacillus casei (Lactobacillus casei), Lactobacillus helveticus carcass (Lactobacillus helveticus), Lactobacillus Homo Hi Ochi (Lactobacillus homohiochii), Lactobacillus pento suspension is any of (Lactobacillus pentosus), Lactobacillus Planta room (Lactobacillus plantarum), Lactobacillus momentum spread (Lactobacillus fermentum) or Lactobacillus brevis (Lactobacillus brevis).

In addition, the plant lactic acid bacteria inoculated in step (d) belong to any one of the genus Lactobacillus and Bifidobacterium It is characterized in that the strain. This strain is preferably a Lactobacillus four K (Lactobacillus sakei), Lactobacillus para casei (Lactobacillus paracasei), Lactobacillus beukeu Nourishing (Lactobacillus buchneri), Lactobacillus brevis (Lactobacillus brevis) or Lactobacillus Coconut Cocos (Lactococcus lactis) It is characterized in that any one of.

The present invention is also characterized in that the food or beverage containing natural gabar prepared according to the above manufacturing method.

The inventors of the present invention have tried various methods to develop fermented products naturally containing Gabba. As a result, it was confirmed that natural gaba can be efficiently obtained when fermenting a raw material comprising glutamic acid using a lactic acid bacterium having a protease generating ability and a gaba generating ability.

The features and advantages of the present invention are summarized as follows.

(1) The present invention is a fermentation product of which the content of Gaba is naturally increased by using lactic acid bacteria having protease activity and vegetable lactic acid bacteria having glutamate decarboxylase activity using various materials including glutamic acid-containing protein as main ingredients. It provides a method of manufacturing.

(2) The fermented product obtained by the method of the present invention can be said to be a fermentation product of a new concept that contains not only the advantages of the lactic acid bacteria fermentation, but also the bioactive substance called gabara, by containing the organic acid and gava produced by the vegetable lactic acid bacteria.

Prior to describing the embodiments, the method of the present invention will be described in detail with each step.

Step (a): Preparation of Medium

According to the method of the present invention, a medium capable of growing a protease-producing lactic acid bacteria is prepared using a component containing a protein including glutamic acid as a component.

Any material used to prepare a medium in the present invention can be used as long as the material containing glutamic acid. Raw materials such as tomatoes and other vegetables and rice bran can be used. If there is a lot of water, it can be used. Raw materials such as rice bran can be extracted with water.

In addition to the main raw material, for the growth of protease-producing lactic acid bacteria, carbon sources, nitrogen sources, trace elements, etc. may be additionally mixed. Available as a carbon source in the medium of the present invention is selected from the group consisting of glucose, sucrose, maltose, fructose, lactose, xylose, galactose, arabinose or a combination thereof, preferably sucrose, fructose Toxins, glucose, galactose, arabinose and lactose, more preferably sucrose, fructose and glucose, most preferably sucrose and glucose. The amount of carbon source used is preferably 1-20% by weight of the natural component, more preferably 1-10% by weight, most preferably 1-4% by weight.

Usable as a nitrogen source in the medium of the present invention is selected from the group consisting of yeast extract, soytone, peptone, bee extract, tryptone, cachetone and combinations thereof, preferably yeast extract, Peptone, tryptone and soyton, more preferably yeast extract and soytone, most preferably yeast extract. The amount of the nitrogen source is preferably 1-20% by weight of the natural component, more preferably 1-10% by weight, most preferably 1-4% by weight.

Usable as trace elements in the medium of the present invention are selected from the group consisting of magnesium sulfate, sodium acetate, manganese sulfate, ferric sulfate, calcium chloride, polysorbate, potassium phosphate and combinations thereof. The amount of the trace element used is preferably 0.001-1% by weight of the natural component, more preferably 0.01-1% by weight, most preferably 0.03-0.5% by weight.

The medium mixed with the main and subsidiary materials is subjected to a sterilization process. Sterilization is preferably carried out in the range 60-130 ° C., more preferably 80 ° C.-121 ° C., preferably for 5-40 minutes, more preferably 5-30 minutes.

Step (b): Lactobacillus Inoculation into Glutamic Acid-Containing Medium

According to a preferred embodiment of the present invention, the lactic acid bacteria which are prepared in the above step and sterilized are inoculated into the medium, and the lactic acid bacteria which can be used in the present invention are limited as long as they can produce glutamic acid by decomposing the protein in the medium using a protease. is not, preferably a Lactobacillus spp, and most preferably Lactobacillus para casei (Lactobacillus paracasei), Lactobacillus casei (Lactobacillus casei), Lactobacillus helveticus carcass (Lactobacillus helveticus), Lactobacillus Homo Hi Ochi (Lactobacillus homohiochii), a Lactobacillus pento suspension (Lactobacillus pentosus), Lactobacillus Plastic tarum (Lactobacillus plantarum), Lactobacillus momentum spread (Lactobacillus fermentum) and Lactobacillus brevis (Lactobacillus brevis).

As a medium for spawning the inoculating bacteria, MRS broth medium is suitable, and the desired bacterial activity can be obtained by incubating at 30 ° C. for 16-48 hours. The amount of lactic acid bacteria inoculated into the medium should be 10 ⁵ -10 ⁹ cfu / g or 10 ⁵ -10 ⁹ cfu / ml after the inoculation, but a lower number of bacteria takes a long time to produce protease. As a result, the incubation time for the production of the product is long, and inoculation of more bacteria is a burden on the production of the spawn.

Step (c): Production of Glutamic Acid-Containing Fermentation

The inoculated medium is evenly mixed and the fermentation proceeds at 20-40 ° C., but fermentation does not easily occur at a temperature below that, and the lactic acid bacteria that are weak in heat cannot be grown to produce a desired fermentation product. The time for culturing the lactic acid bacteria is preferably 8-90 hours, more preferably 12-72 hours, and most preferably 16-48 hours.

Step (d): inoculating the vegetable lactic acid bacteria into the glutamic acid-containing fermentation product

According to a preferred embodiment of the present invention, the lactic acid bacteria are inoculated into the fermentation product containing glutamic acid. The vegetable lactic acid bacteria that can be used in the present invention are glutamic acid contained in the fermentation product using glutamate decarboxylase. It has the ability to convert glutamate to gabba. The lactic acid bacteria used in the present invention are not limited as long as they express glutamate decarboxylase, and are preferably of the genus Lactobacillus and Bifidobacterium . Strain, and most preferably Lactobacillus four K (Lactobacillus sakei), Lactobacillus para casei (Lactobacillus paracasei), Lactobacillus beukeu Nourishing (Lactobacillus buchneri), Lactobacillus brevis (Lactobacillus brevis) and Lactobacillus Coconut Cocos (Lactococcus lactis) to be.

As a medium for spawning the inoculating bacteria, MRS broth medium is suitable, and the desired bacterial activity can be obtained by incubating at 30 ° C. for 16-48 hours.

The inoculum of vegetable lactic acid bacteria inoculated into glutamic acid-containing fermentation should be 10 ⁷ -10 ⁹ cfu / g or 10 ⁷ -10 ⁹ cfu / ml after inoculation. Due to interference by the protease-producing lactic acid bacteria, the second stage of fermentation may not proceed.

Step (e): Production Step of Gaba-Containing Fermentation

The fermented product after the two-stage inoculation is mixed evenly and proceeds to fermentation at 20-40 ℃, and fermentation does not easily occur at temperatures below that, and at a higher temperature, lactic acid bacteria that are weak to heat do not grow to produce a desired fermentation product. none. The time for culturing the lactic acid bacteria is preferably 12-120 hours, more preferably 24-100 hours, and most preferably 48-90 hours.

According to a preferred embodiment of the invention, the Gabba conversion of 50-100% of the glutamic acid produced in the medium, more preferably 70-100%, even more preferably 80-100%, most preferably 90-100% Represents Gaba conversion of.

Gaba-containing fermented product prepared by the present invention includes the organic acid produced by lactic acid bacteria and has the most influence on flavor and acidity of the product and shows various physiological activities.

Hereinafter, the present invention will be described with examples. The following examples are only for illustrating the present invention in more detail, and it will be apparent to those skilled in the art that the scope of the present invention is not limited by these examples according to the gist of the present invention. will be.

For example, the process of preparing tomato fermented tomato fermented products using tomato is as follows. Various tomatoes → crushing → sterilization → (additional ingredients) → protease-producing lactic acid bacteria inoculation → mixing and incubation → further inoculation of vegetable lactic acid bacteria → mixing and culture.

Example 1 Preparation of Media Using Tomato

Using the tomato purchased from the Agricultural Cooperatives three times in cold water to remove dirt and foreign matter on the surface. After 1 kg of washed tomatoes were pulverized using a grinder for 5 minutes, 2% sucrose of tomato weight was added as a carbon source, followed by sterilization at 80 ° C. for 30 minutes to complete the tomato medium. Tomato has a number of advantages as a raw material for the production of natural gaba because a large amount of glutamic acid is present in a free form, or as a component of protein.

Example 2: Preparation of Medium Using Rice Bran

1 kg of rice bran purchased from Nonghyup direct mill (RPC) was washed in 3 L of cold water and then immersed in 10 L of water and extracted at 60 ° C. for 12 hours while stirring at 30 rpm. The extracted rice bran extract was centrifuged to remove solids, and then 3% sucrose and 2% yeast extract were added and sterilized at 121 ° C. for 15 minutes to prepare a medium capable of producing Gabba.

Example 3 Production of Glutamic Acid-Containing Tomato Fermentation

Two tomato medium prepared in the same manner as in Example 1 in a Erlenmeyer flask each containing 1 kg of Lactobacillus casei incubated in MRS broth (deMan Rogosa Sharpe broth) for 18 hours to a final bacterial concentration of 5 × 10 ⁸ cfu / After inoculation to g, incubation for 72 hours was carried out to examine the pH change, protease activity, cell number and glutamic acid content in the culture.

Protease activity measurement was used by modifying the Kunitz method. After lactic acid bacteria having protease activity were cultured in MRS medium, the supernatant was taken and used as a coenzyme solution, and 0.5% casein solution was used as a protein substrate. The coenzyme solution and casein solution were mixed and reacted at 37 ° C. for 1 hour. After completion of the reaction by adding 15% trichloroacetic acid, the supernatant was taken and analyzed by measuring absorbance at 280 nm.

The content of glutamic acid was determined using an amino acid assay modified from RP-HPLC. First, the sample was centrifuged at 8000 rpm for 10 minutes, and then the supernatant was filtered through a membrane filter and diluted to an appropriate concentration. The prepared samples were subjected to RP-HPLC after derivatization with o- phthaldialdehyde (OPA). As an HPLC column, an XTerra column (Waters: RP18 5 mx 4.6 mm x 150 mm) was used.For mobile phase, 0.05 M sodium acetate (pH 7.2) was added as solvent A, and 0.1 M sodium acetate, acetonitrile (HPLC grade) and methanol. (HPLC grade) mixed at 46:44:10 (v / v) (pH 7.2) was used as solvent B. The concentration gradient of the mobile phase was 100% of solvent A. After 30 minutes, solvent B became 100%. After 40 minutes, solvent B was 100%. After 45 minutes, solvent A was 100 again. % And solvent A was adjusted to 100% until 60 minutes later. The flow rate of the mobile phase was fixed at 1 mL / min and clotamic acid was detected by a UV detector at 358 nm.

Table 1 Measurement of changes in cell number, pH and glutamic acid content over time

Fermentation time Cell number (cfu / ml) PH after fermentation Protease Activity
(mg / mg-protein) Glutamic acid content
(%, w / w) 12 hours 3.7 × 10 ⁹ 4.6 0.009 0.25 24 hours 3.6 × 10 ⁹ 4.5 0.013 0.33 36 hours 3.3 × 10 ⁹ 4.5 0.013 0.41 48 hours 2.5 × 10 ⁹ 4.5 0.005 0.42 72 hours 2.3 × 10 ⁹ 4.5 0.001 0.41

Protease activity is expressed as trypsin activity.

Trypsin 1mg / mL: 1800 BAEE U / mg (BAEE: N-Benzoyl-L-Arginine-Ethyl-Ester)

As shown in Table 1, the growth of the inoculated lactic acid bacteria showed almost the highest value after 24 hours, and the change in pH hardly appeared after 24 hours. The pH tends to decrease initially as the glutamic acid contained in the tomato is released, but does not show a large change when the pH is lowered by the organic acid. The content of glutamic acid continues to increase until 36 hours after fermentation, but does not increase after that. The activity of protease increases up to 36 hours of fermentation to produce glutamic acid in tomato medium, after which the protease activity decreases by autolysis and the glutamic acid in the raw material. I think this is because most of them are free.

Example 4 Production of Gaba-Containing Tomato Fermentation

After inoculating the tomato fermentation product subjected to 36 hours fermentation in the same manner as in Example 3, Lactobacillus sakei (KFCC-11321) cultured in MRS broth for 18 hours so that the final bacterial concentration is 3 × 10 ⁸ cfu / g After 72 hours of incubation, the pH change and the Gaba content of the fermentation were examined (Table 2).

The content of Gaba was the same as the analysis of glutamic acid using an amino acid method modified from the RP-HPLC method.

<Table 2> GABA Production of Tomato Fermentation after Lactic Acid Bacteria Supplementation

Additional fermentation time (hrs) pH GABA content
(%, w / v) 0 4.6 0.00 8 4.6 0.01 16 4.33 0.05 24 4.26 0.14 36 4.50 0.26 48 4.90 0.38 60 4.95 0.40 72 4.95 0.40

As shown in Table 2, it was found that glutamic acid naturally produced in tomatoes by protease-producing lactic acid bacteria was converted to Gabba by additionally inoculated Gaba conversion strains. Glutamic acid naturally contained in tomatoes can be seen that the production of Gaba increases logarithmically after 16 hours of fermentation, and the pH increases as the production of Gaba occurs.

Example 5 Production of Glutamic Acid-Containing Rice Bran Ferment

Lactobacillus casei incubated in MRS broth (deMan Rogosa Sharpe broth) for 18 hours containing 1 kg of two rice bran medium prepared in the same manner as in Example 2 in a Erlenmeyer flask, the final bacterial concentration was 5 × 10 ⁸ cfu / After inoculation to g, incubation for 72 hours was carried out to examine the pH change, protease activity, cell number and glutamic acid content in the culture.

TABLE 3 Determination of changes in cell number, pH and glutamic acid content over time

Fermentation time Cell number (cfu / ml) PH after fermentation Protease Activity
(mg / mg-protein) Glutamic acid content
(%, w / w) 12 hours 3.5 × 10 ⁹ 4.5 0.005 0.30 24 hours 3.4 × 10 ⁹ 4.3 0.010 1.33 36 hours 3.5 × 10 ⁹ 4.3 0.014 1.85 48 hours 3.0 × 10 ⁹ 4.4 0.004 1.85 72 hours 2.5 × 10 ⁹ 4.3 0.001 1.86

Protease activity is expressed as trypsin activity.

Trypsin 1mg / mL: 1800 BAEE U / mg (BAEE: N-Benzoyl-L-Arginine-Ethyl-Ester)

As shown in Table 3, the growth of the inoculated lactic acid bacteria showed the highest value after 24 hours, and the change in pH hardly appeared after 24 hours. This result is the same as in Example 3, and does not show a large change in the pH lowered by the organic acid. The content of glutamic acid continued to rise up to 36 hours after fermentation and did not increase thereafter.

Example 6 Production of Gaba-containing Rice Bran Fermentation

After inoculating the rice bran fermentation product which had been fermented for 36 hours in the same manner as in Example 5, lactobacillus sakei (KFCC-11321) incubated in MRS broth for 18 hours so that the final bacterial concentration was 3 × 10 ⁸ cfu / g. After 72 hours of incubation, the pH change and the Gaba content of the fermentation were examined (Table 4).

<Table 4> GABA Production of Rice Bran Fermentation by Additional Inoculation

Fermentation time pH GABA content
(%, w / v) 0 4.42 0.00 8 4.40 0.01 16 4.20 0.06 24 4.19 0.34 36 4.20 0.71 48 4.35 1.00 60 4.38 1.01 72 4.40 1.00

As shown in Table 4, glutamic acid, which is naturally contained in rice bran, was converted to Gabba algebraically when the growth of inoculated Lactobacillus Sakei bacteria reached the end of logarithmic growth, and the Gabba content increased. It was confirmed that the pH in the culture solution increased.

From the above embodiment, using a two-stage fermentation method of vegetable lactic acid bacteria having protease generating ability and gaba generating ability as in the present invention, fermentation of naturally increased Gaba content with a variety of materials containing glutamic acid as a main raw material It is demonstrated that water can be produced effectively.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the same is by way of illustration and example only and is not to be construed as limiting the scope of the present invention. Therefore, the substantial scope of the present invention will be defined by the appended claims and equivalents thereof.

Claims (9)

A method for producing a natural GABA-containing fermented product using two-stage fermentation of vegetable lactic acid bacteria having protease-generating ability and gaba-forming ability, comprising the following steps. (a) preparing a medium containing tomato or rice bran as a raw material containing glutamic acid; (b) Lactobacillus para casei (Lactobacillus paracasei), Lactobacillus casei (Lactobacillus casei), Lactobacillus helveticus carcass (Lactobacillus helveticus), Lactobacillus Homo Hi Ochi (Lactobacillus homohiochii) as a lactic acid bacteria with a protease producing ability in the medium the step of immunization of any one of Lactobacillus pento suspension (Lactobacillus pentosus), Lactobacillus Planta room (Lactobacillus plantarum), Lactobacillus momentum spread (Lactobacillus fermentum) or Lactobacillus brevis (Lactobacillus brevis); (c) culturing the lactic acid bacteria to produce a fermentation product containing glutamic acid; (d) Lactobacillus four K (Lactobacillus sakei) a vegetable lactic acid bacteria with the GABA producing ability to the fermentation, Lactobacillus para casei (Lactobacillus paracasei), Lactobacillus beukeu Nourishing (Lactobacillus buchneri), Lactobacillus brevis (Lactobacillus brevis), or Inoculating any one of Lactococcus lactis ; And (e) culturing the lactic acid bacteria to produce a fermentation product containing gabba. delete delete delete delete delete delete delete delete