JP4628978B2 - Method for producing composition with high content of γ-aminobutyric acid - Google Patents

Description

  The present invention relates to a method for producing a high γ-aminobutyric acid-containing composition using plant juice or plant extract and a food or drink containing the same.

  γ-Aminobutyric acid (hereinafter abbreviated as GABA) is a non-protein constituent amino acid that exists widely in the living world but is known to act as a neurotransmitter in the brain. Yes. GABA as a food material has effective physiological effects for modern people with high health maintenance awareness, such as blood pressure lowering action, tranquilizing action, brain function improving action, climacteric symptom mitigating action, neutral fat increase suppressing action, etc. .

  GABA has been found to be contained in foods such as brown rice, tea, some vegetables, fruits and the like, but its content is low and it is difficult to take an effective amount sufficient to obtain the above-mentioned effects. Therefore, various methods for increasing the GABA content in foods have been studied.

  Methods for increasing the GABA content in foods include inoculating microorganisms having GABA producing ability in foods or increasing the GABA content using enzymes of foods, and extracted GABA produced by microorganisms. There were two kinds of methods of adding to food.

  As for the former method, a method for inoculating a microorganism having a GABA producing ability in food with a medium containing skim milk and tomato juice includes Lactobacillus helveticus and Lactobacillus casei (L. casei). A method of performing lactic acid fermentation by adding two types of lactic acid bacteria (see Patent Document 1), a method of inoculating milk with lactic acid bacteria having glutamate-releasing activity and lactic acid bacteria having glutamate decarboxylase activity (Patent Document 2) Reference), Lactobacillus plantarum (L. plantaram) is added to fish soy sauce and lactic acid fermentation is performed (see Patent Document 3), glutamic acid or glutamic acid-containing material and GABA production ability are added to food and beverage or seasoning food ingredients Method for performing lactic acid fermentation by adding lactic acid bacteria having See Patent Document 4) have been disclosed.

  In addition, as a method for increasing the GABA content using enzymes of foods, a technique for producing GABA-enriched cereals using the action of enzymes originally contained in rice germ, rice bran, wheat germ, etc. (for example, , Patent Documents 5 and 6), a technique for producing a GABA-enriched composition using the action of enzymes contained in vegetables such as tomatoes and pumpkins (see, for example, Patent Documents 7 to 9), tea leaves Techniques for producing tea leaves with a high GABA content by anaerobic treatment (for example, see Patent Document 10) have been reported.

  However, foods with an increased GABA content by these methods are still unsatisfactory although the content is higher than before the treatment. In particular, when adding and mixing to other foods and drinks, it was necessary to further increase the GABA content. In addition, these methods have problems that long-term fermentation is required and productivity is low, and depending on the food, lactic acid bacteria do not necessarily produce GABA in the food. In many of the methods described above, glutamic acid or / and a salt thereof or / and their contents are added, but there is a problem that glutamic acid remains due to poor production efficiency and loses the original flavor of food.

On the other hand, with respect to the latter method, as a method for producing GABA by a microorganism, a method of performing lactic acid fermentation with lactic acid bacteria in a fermentation medium containing glutamic acid and / or a salt thereof can be mentioned. Examples of lactic acid bacteria include Lactobacillus hilgardi K-3 (Patent Document 11), Lactobacillus brevis TY414 (L. brevis TY414) (Patent Document 12), and Lactobacillus sp. Y-3 (Lactobacillus sp. Y-3) (patent document 13) is disclosed. This latter method gives the bad image of adding chemical additives in order to add GABA to the food later. Moreover, the flavor substance derived from a fermentation medium may transfer to a foodstuff and may impair the original flavor of a foodstuff.
Japanese Patent No. 3426157 Japanese Patent No. 3172150 Japanese Patent No. 2704493 JP 2004-215529 A Japanese Patent No. 2590423 JP 2004-159617 A Japanese Examined Patent Publication No. 7-12296 Japanese Patent Publication No. 7-14333 JP 2001-252091 A Japanese Patent No. 3038373 Japanese Patent Laid-Open No. 2003-070462 JP 2000-210075 A JP 2004-357535 A

  From such a background, it is possible to apply to a wide range of foods and efficiently produce GABA by inoculating microorganisms having GABA producing ability in foods or increasing the GABA content using enzymes of foods. A method was sought. The present invention was invented in view of such a situation, and particularly relates to a GABA-rich composition using a plant as a raw material.

The present inventors have carried out intensive investigations and found that the problems described above, with the addition of lactic acid bacteria asparagus juice or Asparagus extract in making enrichment of GABA, proteinogenic other than glutamine and glutamic acid are liberated It has been found that by setting the concentration of each amino acid to a predetermined amount, the production rate of GABA is remarkably increased, and a composition containing a high concentration of GABA can be obtained, and the present invention has been completed.

That is, the first present invention, after the 0.1mM~50mM the abundance of each protein constituent amino acids other than glutamine and glutamic acid contained in the asparagus juice or asparagus extract, lactic acid bacteria with GABA-producing ability And a method for producing a GABA high-content composition characterized by enriching GABA by adding glutamic acid or / and a salt thereof or / and contents thereof and performing lactic acid fermentation. , preferably, the method of the 0.1mM~50mM the abundance of each protein constituent amino acids other than glutamine and glutamic acid the method for adding the yeast extract to asparagus juice or asparagus extract or, exploitation asparagus wherein the GABA high a method of reacting a proteolytic enzyme broth or asparagus extract It is a manufacturing method of the organic composition. Also preferably, the amount of glutamic acid or / and a salt or / and inclusions thereof, a method is 0.1 to 20% by mass of the asparagus juice or asparagus extract, more preferably Is a manufacturing method of the above-mentioned GABA high content composition whose sugar concentration (in Brix conversion) in asparagus juice or asparagus extract is 0.1 to 40% .

  The second aspect of the present invention is a high-GABA composition characterized by being obtained by any one of the production methods described above.

  A third aspect of the present invention is a food or drink characterized by containing the second GABA high-content composition of the present invention.

  According to the present invention, a GABA-rich composition containing GABA at a high concentration and having a small amount of residual glutamic acid can be obtained in a short period of time without adding GABA later.

  Hereinafter, the present invention will be described in detail.

  The plant used in the present invention needs to be asparagus, which is a vegetable that has food experience and can grow lactic acid bacteria in the extract.

Milling asparagus optionally chopped, extracted, compressed, concentrated, solid-liquid separation, heat sterilization, asparagus juice or asparagus by treating sterilized by filtration or the like known techniques alone or two or more in combination An extract is obtained.

  Crushing and shredding are methods of physically crushing plants, crushing by impact, and shredding by cutting. Grinding and shredding may be performed manually using a mortar, kitchen knife, cutter knife, scissors or the like, but an apparatus is used when a large amount of plants are to be treated in a short time. Examples of such an apparatus include a mill, a hammer type pulverizer, a mixer, a blender, and the like, and a vegetable shredding machine may be used. The size of the pulverized and shredded plant body is not particularly limited, but is preferably 2 cm or less, and more preferably 5 mm or less.

  Extraction is an operation of adding a solvent to transfer plant components to the plant. The solvent to be added is most preferably water, but an organic solvent may be used in combination as long as it does not inhibit the growth of lactic acid bacteria. The organic solvent may be used in combination with water at the time of extraction, or the extraction may be performed with the organic solvent alone and then mixed with water. Although the kind of organic solvent is not specifically limited, Ethanol is preferable at the point which can be used for a foodstuff. In the case of water, the temperature of the solvent at the time of extraction is preferably 0 ° C to 100 ° C, more preferably 10 ° C to 80 ° C, and in the case of an organic solvent, -20 ° C to 200 ° C is preferable, and 0 ° C to 120 ° C. ° C is more preferred.

  Squeezing is a method in which a physical pressure is applied to a plant body to squeeze the liquid and transfer the components to the juice. The pressure may be applied only in one direction, may be applied from two or more directions, and may be accompanied by a shearing force. Although the operation of pressing is easy if a commercially available pressing machine is used, it may be performed by a method that does not use a machine such as hand pressing or stepping. At this time, water or hot water may be added to the plant body and squeezed.

  Concentration is an operation of reducing the amount of water and solvent without reducing other components, and may be performed by any method such as concentration under reduced pressure, concentration by heating, concentration using a filtration membrane, but 20 ° C to 60 ° C. It is preferable to carry out vacuum concentration in the range.

  Solid-liquid separation is a method that separates the solvent and components dissolved in it from insoluble solids. For example, any method such as filter filtration, squeeze filtration, centrifugation, or decantation can be used. it can. In order to obtain a clear plant extract, it is preferable to perform filter filtration using a filter aid such as diatomaceous earth.

  Heat sterilization is sterilization by applying heat, and the temperature is preferably 60 ° C to 121 ° C, more preferably 70 ° C to 110 ° C. If it is lower than this temperature range, there is a problem that the effect of sterilization becomes insufficient, and if it is higher than this temperature range, there is a problem that the active ingredient is decomposed.

  Filtration sterilization is an operation for sterilization by passing a membrane of a size through which microorganisms cannot pass under pressure or reduced pressure. The pore diameter of the membrane is preferably from 0.1 to 1.0 μm, more preferably from 0.2 to 0.45 μm.

In the present invention, the asparagus juice or asparagus extract may be used as it is, but it is preferable to use a product from which insolubles have been removed, and particularly preferably, the plant body is crushed, extracted with water, and then sterilized by filtration. It is preferable to use one.

The asparagus juice or asparagus extract obtained by these operations preferably has a sugar concentration (in terms of Brix) of 0.1 to 40% excluding insoluble components, and is preferably 0.1 to 20%. % Is more preferable, and 0.5 to 5% is most preferable. When the sugar concentration (in Brix equivalent) is below this range, the growth of lactic acid bacteria becomes extremely poor, and when the sugar concentration (in Brix equivalent) is above this range, the GABA producing ability of the lactic acid bacteria decreases. When the sugar concentration (converted to Brix) is less than 0.1%, the sugar concentration can be increased. When the sugar concentration (in terms of Brix) exceeds 40%, it is sufficient to add water to reduce it.

The protein-constituting amino acids other than glutamine and glutamic acid in the present invention are 18 kinds of amino acids that normally constitute proteins, excluding glutamine and glutamic acid, that is, asparagine, aspartic acid, serine, threonine, cysteine, glycine , Alanine, methionine, tyrosine, valine, leucine, isoleucine, phenylalanine, proline, tryptophan, histidine, lysine, arginine.

In this invention, it is necessary to make each abundance of protein constituent amino acids other than glutamine and glutamic acid contained in the asparagus juice or asparagus extract prepared as described above 0.1 mM to 50 mM. The abundance of each protein constituent amino acids other than glutamine and glutamic acid at this time does not include those present in the polypeptide or protein, a single does not bind to other molecules in asparagus juice or Asparagus extract Refers only to free molecules present.

In the present invention, the first method of setting the abundance of each of the protein-constituting amino acids other than glutamine and glutamic acid to 0.1 mM to 50 mM is that each amino acid or / and salt thereof, and / or their content is at a predetermined concentration. a method of adding the asparagus juice or asparagus extracts and the like to be.

As protein-constituting amino acids other than glutamine and glutamic acid used in this method, each amino acid may be used separately, but it is preferable to use those containing each amino acid and / or a salt thereof in a balanced manner. Yeast extract, meat extract, fish extract, casamino acid and the like. These are preferable in that they are less expensive than the case of mixing individual amino acids and / or salts thereof. Among these, the yeast extract which is excellent in the effect which improves the GABA production ability of lactic acid bacteria is the most preferable.

  Yeast extract is a water-soluble extract that is extracted by treating the useful components of yeast cells with self-digestion, enzymes, hot water, etc., but it contains water-insoluble components such as cells. It doesn't matter. Any yeast cell may be used as long as it has food experience. Saccharomyces cerevisiae used as beer yeast or baker's yeast, and Candida utilis used as torula yeast. ) Etc. are examples. The yeast extract may be a commercially available product (Difco, Kirin Brewery, Asahi Breweries, etc.) or may be self-adjusted.

By adding yeast extract, the abundance of each protein constituent amino acids other than glutamine and glutamic acid in asparagus juice or Asparagus extract in order to 0.1mM~50mM is asparagus juice or Asparagus extract Although it depends on the origin of the liquid and the production method, it is generally sufficient to add 0.01 to 10 w / w% of yeast extract, preferably 0.1 to 5 w / w%, more preferably 0.5 to 3 w / w%. is there. If the yeast extract concentration falls below this range, the effect is hardly obtained, and if it exceeds this range, the effect cannot be expected to increase, resulting in high costs, adverse effects such as flocking when heated, and a significant impact on taste quality. Occurs.

In the present invention, Futatsume way to 0.1mM~50mM the abundance of each protein constituent amino acids other than glutamine and glutamic acid, added proteolytic enzymes asparagus juice or asparagus extract, is allowed to act, Examples of the method include enzymatic degradation of proteins, polypeptides, and peptides. Examples of the type of enzyme used here include proteases and peptidases that are food additives. The amount of enzyme used to perform enzymatic degradation, the kind of the enzyme, depending on the potency, 0.0001 wt% to 10 wt% based on the solids content of Asparagus juice or Asparagus extract is Preferably, 0.001% by mass to 5% by mass is more preferable. If it is less than this range, there is a problem that sufficient enzymatic degradation cannot be expected, and if it is more than this range, an increase in enzymatic degradation can no longer be expected, and there is a problem that changes the taste quality due to enzymes in the product. is there.

  The temperature during the enzymatic decomposition is preferably 0 ° C to 80 ° C, more preferably 10 ° C to 60 ° C. If it is lower than this range, the progress of the enzymatic reaction tends to be slow and it takes a long time to degrade the protein. If it is higher than this range, the enzyme may be deactivated. Moreover, it is preferable to perform an enzyme deactivation process before adding the lactic acid bacteria of the next process. The temperature of the enzyme deactivation treatment is preferably 50 ° C to 120 ° C, more preferably 70 ° C to 100 ° C. The enzyme deactivation treatment time varies depending on the temperature, but is preferably 5 to 30 minutes, more preferably 5 to 15 minutes.

By the operations described above, contained in the asparagus juice or asparagus extract, after the 0.1mM~50mM the abundance of each protein constituent amino acids other than glutamine and glutamic acid, in the present invention, the GABA-producing ability It is necessary to add the lactic acid bacteria having and glutamic acid or / and its salt or / and their contents. This GABA During lactic acid fermentation asparagus juice or Asparagus extract is performed so that the accumulation by.

  The lactic acid bacteria used in the present invention must be safe for use in foods and have the ability to produce GABA. Examples of such lactic acid bacteria include Lactobacillus brevis, L. et al. hilgardi, L.H. plantaram, L.M. casei, L .; paracasei, L .; helveticus, L.H. bulgaricus, L .; acidophilus, L. et al. sp. Streptococcus lactis, S. et al. Examples include lactic acid bacteria belonging to thermophilus, Enterococcus caseiflavus and the like. Among the lactic acid strains belonging to these, L. brevis UAS-4 (FERM P-20710), L. brevis UAS-6 (FERM P-20711), L.M. brevis IFO 3345, L.M. brevis IFO12005 is more preferable because of its high ability to produce GABA.

Lactic acid bacteria can be added by inoculating a small amount of cells directly into the asparagus juice or asparagus extract prepared as described above, but in order to increase the cell concentration in a short period of time. It is preferable to inoculate a pre-cultured bacterial solution. As the preculture liquid, the same asparagus juice or asparagus extract as in the main culture may be used, or any conventionally known medium can be used. Examples of such a medium include GYP medium generally used for lactic acid bacteria culture, commercially available GAM medium (Nissui Pharmaceutical), MRS medium (Difco), and the like. The amount inoculated with the pre-cultured bacterial solution is 1 / 100,000 of the amount of the main culture medium, preferably 1/1000 to 1/10, and preferably 1/200 to 30 minutes. 1 is more preferable. If the inoculation amount is less than this range, there is a problem that it takes time to increase the bacterial cell concentration, and if it is more than this range, it is already a large scale at the time of pre-culture and there is no need to perform main culture. It is.

  Of the glutamic acid and / or its salt or / and their contents used in the present invention, any glutamate can be used, but it is a food additive and sodium glutamate having excellent solubility in water is preferred. . In addition, any material can be used as the glutamic acid and / or salt thereof, but seasonings such as yeast extract that can be added to foods are preferred.

The amount of glutamic acid or / and a salt or / and inclusions thereof is added to the asparagus juice or Asparagus extract, depending on the origin and production method of asparagus juice or asparagus extract, exploitation asparagus The final concentration of glutamic acid in the juice or asparagus extract is preferably 0.001 to 20 w / w%, more preferably 0.01 to 5 w / w%. Further, glutamic acid and / or a salt thereof and / or their contents may be added in a single amount, or may be added in a plurality of times during fermentation.

  Next, conditions for fermentation will be described.

  The culture temperature during fermentation depends on the strain used, but is 5 ° C to 45 ° C, preferably 15 ° C to 40 ° C, and more preferably 20 ° C to 35 ° C. There is a problem that the growth rate is remarkably inferior even if the culture temperature is higher or lower than this temperature range.

  The pH of the fermentation liquid is preferably adjusted to 4.0 to 6.0, more preferably 4.5 to 5.5, although it depends on the strain used. When the pH is out of this range, there is a problem that the activity of glutamate decarboxylase is lowered and the production rate of GABA is lowered. Any chemical can be used for pH adjustment, such as hydrochloric acid, sulfuric acid, phosphoric acid, nitric acid, acetic acid, butyric acid, lactic acid, formic acid, succinic acid, maleic acid, malic acid, oxalic acid, citric acid, sodium hydroxide, water Examples thereof include potassium oxide, calcium hydroxide, and aqueous ammonia. In the present invention, the pH tends to increase according to the production of GABA, and adjustment is mainly performed by adding an acid. Among these, hydrochloric acid, phosphoric acid, acetic acid, lactic acid, succinic acid, malic acid, Citric acid, more preferably hydrochloric acid, lactic acid, and acetic acid.

  The oxygen condition during fermentation depends on the strain used, but it can be grown under anaerobic or aerobic conditions. However, since there are strains that can grow under aerobic conditions but the GABA production ability decreases, it is preferable to use anaerobic conditions such as anaerobic conditions or moderate stirring, for example, bubbling or rapid stirring is required. There is no.

  The culture time for fermentation is not particularly limited, but is preferably 2 hours to 10 days, more preferably 5 hours to 5 days, and most preferably 8 hours to -3 days. If the fermentation time falls below this range, conversion from glutamic acid to GABA becomes insufficient, and even if this range is exceeded, further effects cannot be expected, and the possibility of contamination and growth of bacteria increases.

  By performing fermentation under the above-mentioned conditions, GABA is enriched and a GABA-rich composition can be obtained. In the production method of the present invention, the GABA high content composition thus obtained can be used as it is as a food or beverage or a seasoned food, but if necessary, sterilization, sterilization, solid-liquid separation It is also possible to carry out operations such as drying to form a clarified liquid or powder. Further, after adding an appropriate carrier, it may be formed into a edible form such as granules, granules, tablets, capsules, gels, pastes, milks, suspensions, liquids, beverages and the like by conventional methods.

  When preparing in a solid form, it is sufficient to use those used in conventional methods such as excipients, binders, disintegrants, lubricants, flavoring agents, colorants, and examples of such carriers. As excipients, lactose, sucrose, glucose, mannitol, sorbit, dextrin, starch, crystalline cellulose, carboxymethylcellulose, hydroxypropylcellulose, dextran, pullulan, anhydrous silicic acid, calcium phosphate, calcium carbonate, calcium sulfate, etc. Binders include crystalline cellulose, sucrose, mannitol, dextrin, hydroxypropylcellulose, hydroxypropylmethylcellulose, polyvinylpyrrolidone, gum arabic, dextran, pullulan, water, ethanol, etc., and disintegrants include starch, carboxymethylcellulose, hydroxypropyl Pyrcellulose, dextrin, crystalline cellulose, etc., as a lubricant, stearic acid and its metal salts, talc, boric acid, fatty acid sodium salt, sodium lauryl sulfate, magnesium lauryl sulfate, silicic acid anhydride, etc. Examples include sucrose, orange peel, citric acid, tartaric acid and the like.

  When preparing in a liquid state, it is sufficient to use an emulsifier, a solubilizer, a dispersant, a suspending agent, a thickener, a buffer, a stabilizer, a flavoring agent, etc. As the emulsifier and solubilizer, lecithin, sucrose fatty acid ester, sorbitan fatty acid ester, sodium lauryl sulfate and the like, and as the dispersant and suspending agent, lecithin, sucrose fatty acid ester, methylcellulose, gum arabic, gelatin and the like, As a thickener, methylcellulose, carboxymethylcellulose, gum arabic, gelatin, etc., as a buffer, citrate, succinate, etc., as a stabilizer, lecithin, gum arabic, gelatin, methylcellulose, as a flavoring agent What was mentioned above can be illustrated.

  In addition, sugars, sugar alcohols, salts, oils and fats, amino acids, organic acids, fruit juices, vegetable juices, fragrances, spices, alcohols, glycerin, and the like can be added to improve the taste. In addition, hydrochloric acid, sulfuric acid, phosphoric acid, acetic acid, butyric acid, lactic acid, succinic acid, malic acid, oxalic acid, citric acid, sodium hydroxide, potassium hydroxide, calcium hydroxide, aqueous ammonia, etc. are added for pH adjustment. can do.

  Next, the 3rd food / beverage products of this invention are demonstrated.

  The food / beverage products of this invention are characterized by containing the GABA high content composition manufactured as mentioned above. The content of the GABA high-content composition in the food or drink is not particularly limited, but the GABA high-content composition itself can be used as a food or drink, but the amount consumed per day is generally converted to GABA. It is preferable to mix | blend so that it may become 10-500 mg. If it is less than this range, the effect may not be expected, and if it is more than this range, the increase in the effect may no longer be expected.

  When a GABA high content composition is included in an existing food or drink or seasoning food, it is not particularly limited as a base food or drink or seasoning food. For example, processed noodles such as udon and pasta, meat such as ham and sausage Processed foods, processed fishery products such as kamaboko and chikuwa, processed milk products such as butter, milk powder and fermented milk, desserts such as jelly and ice cream, processed foods such as breads, confectionery and seasonings, and refreshing Drinks such as drinking water, alcohols, fruit juice drinks, vegetable juice drinks, milk drinks, carbonated drinks, coffee drinks, alcohols and the like are preferred.

  The form of the GABA high-content composition of the present invention to be included in a food or beverage or seasoning food is not particularly limited, and liquids are used for beverages, gummi, candy, etc., and powders are used for tablets, granules, capsules, etc. Can be used.

EXAMPLES The present invention will be specifically described below with reference to examples, but the present invention is not limited to these examples. In the examples, amino acids were analyzed by the following method.
That is, it measured on the following conditions by the high performance liquid chromatography method (HPLC method), and detected using the fluorescence detector.
HPLC: Shimadzu Corporation LC-9A
Column: Shim-pack ISC-07 / S1504
Mobile phase: 0.2 N sodium citrate buffer (pH 2.2)
Flow rate: 0.3 ml / min Temperature: 55 ° C
Reaction solution: ortho-phthalaldehyde Detection wavelength: excitation wavelength 348 nm, fluorescence wavelength 450 nm

Example 1 [Lactic acid bacteria L. GABA production by brevis UAS-4 (FERM P-20710) and the effect of addition of glutamate and yeast extract on it]
1 L of water was added to each 1 kg of asparagus, tomato, apple, and mandarin orange and pulverized with a household mixer. Diatomaceous earth was added to the crushed liquid, and suction filtration was performed using a filter paper (ADVANTEC Toyo No. 5C) to obtain a clear filtrate. Each filtrate was concentrated or diluted with water to adjust Brix to 3.0%. 1 L of each raw material filtrate was divided into 5 equal parts, 1% w / w% of yeast extract was added to 2 sections, and suction filtration sterilization was performed using filter paper having a pore diameter of 0.20 μm. Thereafter, 3.0 w / w% sodium glutamate in terms of glutamic acid was added to the 1st district with added yeast extract and the 1st district with no yeast extract added. Among the 5 sections prepared in this way, L. Brevis UAS-4 was inoculated. Table 1 summarizes each treatment area.

  The preculture was performed as follows. 10 ml of FYP medium in which the glucose in the GYP medium was changed to fructose was inoculated with the strain that had been cryopreserved at −80 ° C., and the culture was allowed to stand at 30 ° C. for 24 hours.

Then, stationary culture was performed at 30 degreeC for 72 hours, and each fermentation liquid was obtained. The result of the GABA content of each plant extract after 24, 48, and 72 hours and the fermented product of each process section is shown in FIGS.

  As a result, in all plant extracts, the addition of sodium glutamate significantly increases the GABA content, and the addition of yeast extract increases the rate of GABA production and further increases the GABA content (FIGS. 1 to 3). In the group where both sodium glutamate and yeast extract were added, the asparagus extract had the highest GABA content, producing approximately 92% of the theoretically possible GABA amount after 48 hours and approximately 98% after 72 hours. (Figs. 2 and 3).

Example 2 [Lactic acid bacteria L. Breba IFO12005 GABA production and the effect of adding glutamate and yeast extract on it]
The lactic acid bacteria used in Example 1 were L. The experiment was conducted in the same manner except that the Brevis IFO12005 was used. The result of the GABA content of each plant extract after 24, 48, and 72 hours and the fermented product of each process section is shown in FIGS.

  As a result, L. brevis IFO12005 also has an effect of promoting GABA production of yeast extract. In the group to which both sodium glutamate and yeast extract are added, GABA content is the highest in the asparagus extract and can be theoretically produced after 48 hours. Approximately 86% of the amount is produced, approximately 94% after 72 hours (FIGS. 4 and 5). From this, the GABA production promoting effect of yeast extract is L. It turned out that it is not an effect peculiar to brevis UAS-4.

Example 3 [Lactic acid bacteria L. asparagus extract Effect of GABA production and glutamate concentration by brevis UAS-4 (FERM P-20710)]
In Example 1, the plant raw material was only asparagus, and the experiment was performed again by adding sodium glutamate concentrations of 1.0, 3.0, and 5.0 w / w% in terms of glutamic acid. The results of the glutamic acid concentrations after 24, 48 and 72 hours and the GABA content of the fermented product in each treatment zone are shown in FIGS.

  As a result, in the group to which only sodium glutamate was added, even when the amount of glutamic acid added was high, GABA production reached a peak, and much glutamic acid remained. On the other hand, in the group where both sodium glutamate and yeast extract were added, the GABA content was directly proportional to the amount of glutamic acid added, and the addition of yeast extract also improved the total production of GABA (FIGS. 7-9).

Example 4 [Lactic acid bacteria L. asparagus extract effect of each concentration of protein constituent amino acids free on GABA production by brevis UAS-4 (FERM P-20710)]
Asparagus clarified filtrate containing 2.0% Brix was obtained in the same manner as in Example 1. The amino acid analysis of this filtrate was as shown in Table 2.

After adding the seven amino acid compositions shown in Table 3 to this extract, autoclaving was performed.

Thereafter, sodium glutamate monohydrate was converted to glutamic acid at a final concentration of 1 w / w%, lactic acid bacteria L. A pre-culture solution of brevis UAS-4 (FERM P-20710) was added to 1 v / v% and cultured at 30 ° C. for 48 hours. Table 4 shows the GABA content (g / L) of each section after culture.

As a result, GABA production was promoted in the group where each amino acid of protein constitution except glutamine and glutamic acid was added (2nd group) and in the group where less Cys, Met, Ile, Tyr, Phe was added to the asparagus extract (3rd group). In the group having a low concentration of one or more protein constituting amino acids (1, 4 to 8 group), almost no GABA production occurred. From this, it was found that it is effective for GABA production to make the concentration of the protein-constituting amino acids excluding glutamine above a certain level.

Example 5 (Production of GABA high content composition)
The fermented liquor from 1 to 5 in Table 1 of each plant obtained in Example 1 was treated at 100 ° C. for 5 minutes and filtered through a filter (Millipore, pore size 0.20 μm). Twenty panelists drank 20 kinds of GABA-rich beverages obtained in this way, and their taste and ease of drinking were evaluated on a 5-point scale (1 to 5 points). The easier the score is). The average score is shown in Table 5.

As a result, in any plant, the fermentation broth added with glutamic acid alone has a low score, and the others are generally high in score, and the deterioration of the taste of the fermentation broth due to the large amount of glutamic acid is eliminated by the addition of yeast extract. I understood.

Example 6 [Production of food containing high GABA content]
180 ml of the asparagus fermentation solution obtained in Example 5 was freeze-dried to obtain 11.9 g of a brown powder. This was mixed with medium-strength flour so as to be 1 w / w% to prepare udon noodles. When the udon noodles were cooked, they were good noodles with a slight sweetness and a refreshing scent.

In Example 1 of this invention, the GABA content in the fermented liquor of 1st zone-5th zone in Table 1 of each plant. 24 hours later. In Example 1 of this invention, the GABA content in the fermented liquor of 1st zone-5th zone in Table 1 of each plant. 48 hours later. In Example 1 of this invention, the GABA content in the fermented liquor of 1st zone-5th zone in Table 1 of each plant. 72 hours later. In Example 2 of this invention, the GABA content in the fermented liquor of 1st ward to 5th ward in Table 1 of each plant. 24 hours later. In Example 2 of this invention, the GABA content in the fermented liquor of 1st ward to 5th ward in Table 1 of each plant. 48 hours later. In Example 2 of this invention, the GABA content in the fermented liquor of 1st ward to 5th ward in Table 1 of each plant. 72 hours later. The GABA content in the fermented liquor in 1st to 5th districts in Table 1 at each glutamic acid concentration in Example 3 of the present invention. 24 hours later. The GABA content in the fermented liquor in 1st to 5th districts in Table 1 at each glutamic acid concentration in Example 3 of the present invention. 48 hours later. The GABA content in the fermented liquor in 1st to 5th districts in Table 1 at each glutamic acid concentration in Example 3 of the present invention. 72 hours later.

Claims (7)

After the abundance of each protein constituent amino acids other than glutamine and glutamic acid contained in the asparagus juice or Asparagus extract and 0.1MM～50mM, the lactic acid bacteria with γ- aminobutyric acid-producing ability, glutamic acid or / and The manufacturing method of the high content composition of (gamma) -aminobutyric acid characterized by enriching (gamma) -aminobutyric acid by adding the salt or / and those content, and performing lactic acid fermentation. How to 0.1mM~50mM the abundance of each protein constituent amino acids other than glutamine and glutamic, asparagus juice or Asparagus extract is a method of adding a yeast extract according to claim 1, wherein the γ- amino A method for producing a composition having a high butyric acid content. How to 0.1mM~50mM the abundance of each protein constituent amino acids other than glutamine and glutamic, asparagus juice or Asparagus extract is a method for applying a proteolytic enzyme according to claim 1, wherein the γ- A method for producing a composition containing a high amount of aminobutyric acid. The amount of addition of glutamic acid or / and a salt or / and inclusions thereof, according to any of claims 1 to 3 from 0.1 to 20 wt% with respect to asparagus juice or Asparagus extract A method for producing a high γ-aminobutyric acid-containing composition. The method for producing a high γ-aminobutyric acid-containing composition according to any one of claims 1 to 4, wherein the sugar concentration (converted to Brix) in the asparagus juice or asparagus extract is 0.1 to 40%. A γ-aminobutyric acid-rich composition obtained by the production method according to claim 1. A food / beverage product comprising the high γ-aminobutyric acid-containing composition according to claim 6.