JP2008017703A - METHOD FOR PRODUCING FOOD COMPRISING gamma-AMINOBUTYRIC ACID AND ORNITHINE - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a method by which both GABA (γ-aminobutyric acid) and ornithine can simultaneously and efficiently be produced, to provide a method for efficiently increasing the GABA and ornithine contents in a food without adding a pure substance of each or/and a salt thereof or/and a material containing the same and to provide a composition and a food and drink reasonably affording continuous ingestion of necessary amounts of the GABA and ornithine by the method. <P>SOLUTION: A lactic acid bacterium having both the ability to produce the GABA and the ability to produce the ornithine is inoculated into a culture medium or a food raw material containing glutamic acid or/and salts thereof and arginine or/and salts thereof. Thereby, lactic acid fermentation is carried out. <P>COPYRIGHT: (C)2008,JPO&INPIT

Description

  The present invention relates to a method for producing a composition or food containing γ-aminobutyric acid and ornithine obtained by lactic acid bacteria culture.

  γ-aminobutyric acid (hereinafter abbreviated as GABA) is a non-protein free amino acid that exists widely in the living world but is known to act as a neurotransmitter in the brain. It has been. GABA as a food material has physiological actions such as a blood pressure lowering action, a tranquilizing action, a brain function improving action, a climacteric disorder symptom reducing action, a growth hormone secretion promoting action, and a neutral fat increase inhibiting action.

  On the other hand, ornithine is a non-protein free amino acid widely present in a small amount in the living world, and is known to be involved in the metabolism of ammonia as an important component of the ornithine cycle in the liver. Ornithine as a food material has physiological actions such as growth hormone secretion promoting action, liver function improving action, immune function improving action, fatigue improving action, and skin beautifying action.

  From this, it is expected that a composition containing GABA and ornithine has many physiological effects that are effective for modern people with high health maintenance awareness.

  Conventionally, when a composition containing GABA and ornithine is to be produced, each pure product and / or salt thereof and / or their contents are mixed, or the other amino acid is produced in the production process of one amino acid. The pure product and / or the salt thereof and / or the content thereof must be added to increase the cost.

  In addition, GABA is known to be contained in foods such as brown rice, tea, some vegetables, fruits, etc., 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 types of methods of adding to the food.

  As for the former method, a method of inoculating a microorganism having a GABA producing ability into foods is carried out by adding Lactobacillus helveticus and Lactobacillus casei to a medium containing skim milk and tomato juice. A method of performing lactic acid fermentation by adding two kinds 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, although the foodstuff which raised GABA content by these methods was high compared with the thing before a process, it was still unsatisfactory. 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.

  On the other hand, ornithine is also known to be relatively contained in mushrooms, rainbow trout, flounder, yellowfin tuna, cheese, etc., but its content is low and it is difficult to ingest an effective amount sufficient to obtain the above-mentioned effects. there were.

  For example, Non-Patent Document 1 includes 284.4 mg in 100 g for the mature pattern of Shirotamogitake (currently Bunashimeji), and Patent Document 14 includes 758 mg in 100 g of lyophilized extract of shijimi that has been slowly cooled at −20 ° C. Patent Document 2 describes that 44.62 mg is contained in 100 g of Chinese cabbage preserved at 7 ° C. for 30 days, and Non-Patent Document 3 includes 52.8 mg in 100 g of Mini St Paulin type cheese. However, it was hardly contained in foods other than these, and there were few foods in which the amount of ornithine in solid content 100g exceeded 50 mg.

  There are two methods for increasing the ornithine content in foods, a method for increasing the ornithine content using the biological reaction of organisms containing ornithine and a method for adding ornithine produced industrially to foods.

  As a method for increasing the ornithine content by utilizing the biological reaction of an organism containing ornithine, a method is known (Patent Document 14) in which a cooling load is applied to swordfish shells to promote the ornithine biosynthesis reaction. However, this method can only be applied to Shijimi.

  As a method for industrially producing ornithine, there are a chemical synthesis method and a fermentation method. However, this method gives a bad image of adding a chemical additive in order to add ornithine to the food later.

The GABA content in the above-mentioned food containing a relatively large amount of ornithine is not included in Shijimi and Mini St Paulin type cheeses. Only 12.42 mg is contained in 100 g. Therefore, in order to ingest GABA and ornithine from foods, they must be ingested from different foods, and since the content thereof is small, it is unrealistic to ingest it to the extent that its effect can be expected.
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 Patent No. 1984200 Japanese Patent No. 1989077 JP 2001-252091 A Japanese Patent No. 3038373 Japanese Patent Laid-Open No. 2003-070462 JP 2000-210075 A JP 2004-357535 A JP 2001-204432 A Japanese Journal of Food Industry Vol.31 No.10 (1994), 649-655 Japanese Society of Food Industry Vol.33 No.10 (1996), 701-707 Int. Diary J. , 8 (1998), 889-898

  From such a background, a method capable of efficiently producing both GABA and ornithine simultaneously has been demanded. Moreover, the method of increasing GABA and ornithine content in a foodstuff efficiently without adding each pure product or / and their salts or / and those content was calculated | required. The present invention has been invented in view of such a situation.

  As a result of intensive studies on the above problems, the present inventors have found that the above problems can be solved by using a lactic acid bacterium having both γ-aminobutyric acid-producing ability and ornithine-producing ability, and have completed the present invention. .

  That is, in the first aspect of the present invention, lactic acid fermentation is performed by inoculating a medium containing glutamic acid or / and salts thereof and arginine or / and salts thereof with lactic acid bacteria having both γ-aminobutyric acid producing ability and ornithine producing ability. The gist of the present invention is a method for producing a composition containing γ-aminobutyric acid and ornithine.

  In addition, the second of the present invention is a food containing γ-aminobutyric acid and ornithine characterized by inoculating a lactic acid bacterium having both γ-aminobutyric acid producing ability and ornithine producing ability into a food raw material and fermenting lactic acid. The manufacturing method is the gist of the present invention, preferably the above-described method for producing a food containing γ-aminobutyric acid and ornithine, which further contains glutamic acid or / and a salt thereof in the food material, and preferably food. The method for producing a food containing γ-aminobutyric acid and ornithine according to any one of the above, wherein the raw material further contains arginine or / and a salt thereof, and preferably the food raw material is previously treated with a proteolytic enzyme. A method for producing a food containing γ-aminobutyric acid and ornithine as described above, and more preferably γ The method for producing a composition or food containing γ-aminobutyric acid and ornithine according to any one of the above, wherein the lactic acid bacterium having both aminobutyric acid producing ability and ornithine producing ability is a lactic acid bacterium belonging to Lactobacillus brevis (Lactobacillus brevis) More preferably, it is a method for producing a food containing γ-aminobutyric acid and ornithine as described above, wherein the food material is obtained from asparagus.

  Furthermore, a third aspect of the present invention is a composition or food containing γ-amino acid and ornithine produced by any one of the production methods described above.

  According to the present invention, both GABA and ornithine can be efficiently produced simultaneously. In addition, the GABA and ornithine contents in the food can be efficiently increased without adding each pure product or / and their salts or / and their contents.

  Hereinafter, the present invention will be described in detail. First, the first of the present invention will be described.

  The lactic acid bacteria used in the present invention are safe even when used in foods, and must have both GABA producing ability and ornithine producing ability. Examples of such lactic acid bacteria include Lactobacillus brevis, L. et al. and lactic acid bacteria belonging to lactis and the like. Among the lactic acid strains belonging to these, L. has previously been found from nature by the present inventors. brevis UAS-4 (FERM P-20710), L.M. brevis UAS-6 (FERM P-20711) is more preferred because of its high ability to produce GABA and ornithine. In the present invention, lactic acid strains belonging to these may be used alone, or two or more strains may be used in combination.

  In the present invention, any conventionally known medium can be used as a medium for growing lactic acid bacteria and causing GABA production and ornithine production. As such a medium, a GYP medium (D-glucose 1%, peptone 0.5%, yeast extract 1%, sodium acetate trihydrate 0.2%, twin 80 0.05%, sulfuric acid, which is commonly used for lactic acid bacteria culture, is used. Magnesium heptahydrate 0.02%, manganese sulfate tetrahydrate 10 ppm, iron sulfate heptahydrate 10 ppm, sodium chloride 10 ppm), FYP medium (GYP medium sugar converted to D-fructose), Commercially available GAM medium (Nissui Pharmaceutical), MRS medium (Difco), etc. can be used, and yeast extract, meat extract, malt extract, fish meat extract, soybean degradation product, peptone, polypeptone, potato exudate, etc. alone Alternatively, a medium in which two or more kinds are dissolved in water can also be used.

  In the present invention, it is necessary that glutamic acid or / and salts thereof and arginine or / and salts thereof are contained in the medium described above. Examples of salts of glutamic acid include sodium salt, potassium salt, magnesium salt, and calcium salt. As a method for containing glutamic acid or / and salts thereof, a medium containing glutamic acid or / and salts thereof may be included in the medium. Examples of such inclusions include acid hydrolysates of proteins, enzyme degradation products, yeast extracts, and the like. Among them, yeast extracts are preferable because they have a high effect of promoting the ability to produce GABA of lactic acid bacteria.

  The addition amount of glutamic acid or / and salts thereof is 0.1% by mass to 40% by mass, preferably 0.2% by mass to 25% by mass, and preferably 1% by mass to 20% by mass with respect to the medium in terms of glutamic acid. Further preferred. If the amount is less than this range, the amount of GABA accumulated in the culture solution is reduced. If the amount is more than this range, glutamic acid and / or glutamate remains, or even if the whole amount is converted to GABA, it takes a long time to culture. There is a case. Glutamic acid or / and salts thereof may be added in total before culturing, or may be added in a plurality of times during the culturing period. In terms of workability, it is preferable to add the whole amount at one time, but in order to avoid a sudden change in pH, it can also be added in small amounts, in which case it is preferably 2 to 10 times, more preferably 2 to 5 times.

  Examples of arginine salts include hydrochloride and glutamate. As a method for containing arginine or / and a salt thereof, a content containing arginine or / and a salt thereof may be included in the medium. Examples of such inclusions include acid hydrolysates, enzyme degradation products, yeast extracts and the like of proteins. Among them, yeast extracts are preferable because they have a high effect of promoting ornithine production ability of lactic acid bacteria.

  The addition amount of arginine or / and salts thereof is 0.1% by mass to 40% by mass, preferably 0.2% by mass to 25% by mass, and preferably 1% by mass to 20% by mass with respect to the medium in terms of arginine. Further preferred. If it is less than this range, the amount of ornithine accumulated in the culture solution will be small, and if it is more than this range, arginine and / or arginine salt will remain, or even if the whole amount is converted to ornithine, it will take a long time for cultivation. There is a case. Arginine or / and salts thereof may be added in total before culturing or may be added in a plurality of times during the culturing period. In terms of workability, it is preferable to add the whole amount at one time, but in order to avoid a sudden change in pH, it can also be added in small amounts, in which case it is preferably 2 to 10 times, more preferably 2 to 5 times.

  Next, the culture conditions in the present invention will be described.

  The method of adding lactic acid bacteria may be a method of directly inoculating a small amount of bacterial cells in the above-mentioned medium, or a method of inoculating a pre-cultured bacterial solution in order to increase the bacterial cell concentration in a short period of time. As the preculture solution, the above-mentioned medium can be used as in the case of the main culture. 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.

  Although the temperature at the time of culture | cultivation is based also on the strain to be used, it is 5 to 45 degreeC, Preferably it is 15 to 40 degreeC, More preferably, it is 20 to 35 degreeC. 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 culture solution depends on the strain to be used, but is preferably adjusted to 3.5 to 8.0, more preferably 4.0 to 7.0, and 4.5 to 6.5. It is most preferable to adjust. 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.

  Oxygen conditions during culture depend on the strain used, but 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.

  By culturing as described above, GABA and ornithine are accumulated in the culture broth, and the culture broth can be used as the composition containing GABA and ornithine in the present invention. Moreover, the culture supernatant obtained by removing the cells from this culture solution can be used as a composition containing GABA and ornithine in the present invention. Further, the culture supernatant may be concentrated and dried, or the culture supernatant may be subjected to treatment such as crystallization and ion exchange to improve the purity of GABA and / or ornithine.

  The cells can be removed by a conventionally known method. For example, a filter press, a Buchner funnel, a centrifugal filter, a centrifuge, a decanter, or the like can be used.

  In order to concentrate the culture supernatant, conventionally known concentrators can be used, and examples include evaporators, concentration cans, evaporator (Okawara Seisakusho), wall wetter (manufactured by Kansai Chemical Machinery), and the like.

  In order to dry the culture supernatant, a conventionally known drying apparatus can be used. For example, a vacuum freeze dryer, a vacuum shelf dryer, a hot air dryer, a vacuum kneader, a ribocorn dryer, a drum dryer, an oscillatory flow Examples include dryers and spray dryers. The liquid to be dried may be the culture supernatant as it is, or may be a concentrated product, but a solid concentration of 5 to 70% by mass, preferably 5 to 50% by mass, more preferably 10 to 40% by mass is used. It is preferable to do.

  What was dried as mentioned above can also be pulverized into powder. A conventionally known pulverizer can be used for the pulverization, and examples thereof include a mortar, a blender mixer, a cutter mill, a hammer mill, a jet mill, and a feather mill.

  The composition containing GABA and ornithine obtained as described above contains GABA in an amount of 5% by mass or more, preferably 10% by mass or more, and ornithine in an amount of 0.1% by mass or more when dry. Contained, preferably 0.5% by mass or more.

  In order to further improve the concentration of the produced GABA and / or ornithine, further purification may be performed, and examples of such purification methods include crystallization, ultrafiltration, ion exchange, activated carbon treatment, and chromatographic separation. Among these, ultrafiltration, ion exchange, and activated carbon treatment are preferable.

  For purification by crystallization, conventionally known methods can be used, and examples thereof include concentration crystallization, cooling crystallization, heating dissolution / cooling crystallization, isoelectric crystallization, and crystallization method in which an organic solvent is added. Since GABA and ornithine have different solubility and isoelectric point, the purity of one amino acid can be arbitrarily increased by utilizing the difference. The precipitated crystals may be separated by a conventionally known method. For example, a centrifugal filter, a Buchner funnel, a filter press or the like can be used.

  Purification by ultrafiltration can remove high molecular weight components, and the molecular weight cutoff can be selected so as to achieve the desired GABA and ornithine purity. The molecular weight cutoff of the ultrafiltration membrane is preferably 1000-200000, more preferably 3000-10000. If it is lower than this range, there is a problem that purification takes time, and if it is higher than this range, there is a problem that the purity of GABA and ornithine is not improved.

  Although ion exchange can use a conventionally well-known method, Preferably it is the refinement | purification using an ion exchange resin tower. As the ion exchange resin, either a cation exchange resin or an anion exchange resin can be used. For example, when a cation exchange resin is used, a method in which a culture solution is passed through an acid-converted H-form to adsorb GABA and ornithine to the resin, and an alkali is passed through to elute GABA and ornithine is preferred. used. In addition, for example, when an anion exchange resin is used, a method in which a culture solution is passed through an alkali-converted OH type to adsorb GABA and ornithine to the resin, and an acid is passed through to elute GABA and ornithine is preferred. Used for. As the ion exchange resin, a cation exchange resin is preferable, and a strong acid cation exchange resin is particularly preferable. In addition, since GABA and ornithine have different affinity for ion exchange resins, the purity of one amino acid can be arbitrarily increased by fractionating and collecting the eluent over time.

  Conventionally known methods can be used for the activated carbon treatment, such as a batch method in which activated carbon is put into a culture solution containing GABA and ornithine and stirred, and a column method in which activated carbon is packed into a column and the culture solution is passed therethrough. Either method is acceptable.

  Moreover, the composition containing GABA and ornithine produced by the present invention can be added as a raw material to an existing food or beverage or seasoning food. Although it is not particularly limited as a base food or drink or seasoned food, for example, processed noodles such as udon and pasta, processed meat foods such as ham and sausage, marine processed foods such as kamaboko and chikuwa, butter, powdered milk, fermented milk, etc. Processed milk products, desserts such as jelly and ice cream, processed foods such as breads, confectionery, seasonings, and soft drinks, alcohol, fruit juice beverages, vegetable juice beverages, milk beverages, carbonated beverages, Beverages such as coffee drinks and alcohols are preferred.

  The content of the composition containing GABA and ornithine in such a food or drink or seasoning is not particularly limited, but is formulated so that the amount taken per day in terms of GABA is 10 to 3000 mg. Or it is preferable to mix | blend so that the quantity taken per day in conversion to ornithine may be 10-5000 mg. If the amount is less than this range, the effect may not be expected, or the food or seasoning may be excessively consumed to obtain the effect. If the amount is more than this range, the effect may not be expected any longer. There is.

  Next, the second invention of the present invention will be described. As the lactic acid bacteria having both GABA producing ability and ornithine producing ability, those described in the first aspect of the present invention are used in the same manner.

  The raw material for food used in the present invention may be any material as long as lactic acid bacteria can grow. Examples include vegetables, fruits, seeds, cereals, potatoes, beans, algae, mushrooms, and seafood. , Meat, milk, eggs and the like. Among these, asparagus is preferable in that it has a high effect of enhancing the GABA production performance of lactic acid bacteria.

  These various food materials may be used as they are, or as long as the effects of the present invention are not impaired, known techniques such as washing, grinding, shredding, extraction, pressing, concentration, solid-liquid separation, heat sterilization, and filter sterilization are used alone. Alternatively, it may be used after processing by combining two or more. Moreover, you may perform operations, such as adjustment of a water content, pH adjustment, sugar addition, and addition of the component used for an above-described culture medium as needed.

  The food material obtained by these operations preferably has a sugar concentration (in terms of Brix) of 0.1 to 40%, more preferably 0.1 to 20%, excluding insoluble components. 0.5 to 5% is most preferable. When the sugar concentration (Brix equivalent) is below this range, the growth of lactic acid bacteria becomes extremely poor, and when the sugar concentration (Brix equivalent) is above this range, the GABA producing ability and ornithine producing ability of the lactic acid bacteria are reduced.

  Further, in the present invention, in order to increase the GABA and / or ornithine content, the glutamic acid or / and salt thereof, or arginine or / and salt thereof described in the first of the present invention is added to the prepared food material. be able to. Only one or both of glutamic acid or / and salts thereof and arginine or / and salts thereof may be added. Further, the addition amount is as described in the first of the present invention.

  Moreover, in this invention, in order to increase GABA and / or ornithine content, the adjusted foodstuff raw material can also be enzyme-processed. This enzyme treatment increases the content of free glutamic acid and free arginine by treating proteins and peptides contained in food ingredients with each degrading enzyme, or converts glutamine to glutamic acid by glutaminase and increases the content of free glutamic acid There is a meaning. In the present invention, only one type of the above-described enzyme may be used, or two or more types may be used simultaneously or in separate steps. The amount of the enzyme to be used is preferably 0.01% by mass to 10% by mass and more preferably 0.1% by mass to 5% by mass with respect to the protein in the adjusted food or food material. . If it is less than this range, the effect of adding an enzyme can hardly be expected, and if it exceeds this range, no further effect can be expected.

  As the culture conditions in the present invention, the same conditions as those described above in the first aspect of the present invention can be employed.

  By culturing as described above, GABA and / or ornithine accumulates in the raw material of the food and can be used as it is as a food containing GABA and ornithine in the present invention. It is also possible to carry out operations such as sterilization, solid-liquid separation, and drying to form a clarified liquid or powder.

  The content of GABA and ornithine in the food containing GABA and ornithine obtained as described above is not particularly limited, but is formulated so that the amount taken per day in terms of GABA is 10 to 3000 mg. Or it is preferable to mix | blend so that the quantity taken | dosed per day in conversion to ornithine may be 10-5000 mg. If the amount is less than this range, the effect may not be expected, or the food needs to be excessively consumed to obtain the effect, and if it is more than this range, the increase in the effect may no longer be expected.

  The composition containing GABA and ornithine obtained according to the first aspect of the present invention and the form of food containing GABA and ornithine obtained according to the second aspect of the present invention are prepared by adding a suitable carrier and then granulating by a conventional method. It may be formed into an edible form such as a shape, granule, tablet, capsule, gel, paste, milk, suspension, liquid, or beverage.

  When adjusting to a solid, what is used in conventional methods such as excipients, binders, disintegrants, lubricants, flavoring agents, and coloring agents may be used, and examples of such carriers As an excipient, 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 adjusting to a liquid state, an emulsifier, a solubilizer, a dispersant, a suspending agent, a thickening agent, a buffering agent, a stabilizer, a flavoring agent, etc. may be used, 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.

In the present invention, the content of amino acids including GABA is a value determined 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-10A
Column: Shim-pack Amino-Li (100 mm L. × 6.0 mm ID)
Mobile phase: Amino acid mobile phase kit Li form Gradient elution flow rate: 0.6 ml / min Column temperature: 39 ° C
Reaction solution: ortho-phthalaldehyde (post column)
Reaction liquid rate: 0.3 ml / min Reaction temperature: 39 ° C.
Detection wavelength: excitation wavelength 350 nm, fluorescence wavelength 450 nm

  Hereinafter, the present invention will be specifically described with reference to examples, but the present invention is not limited thereto. The content of amino acids including GABA in this example was measured by the method described above.

Example 1
To 1 L of GYP medium containing 5.0% by weight sodium glutamate and 1.0% by weight arginine, 10 mL of a cell suspension of Lactobacillus brevis UAS-4 pre-cultured and in a steady state was added. The culture was started at 30 ° C. under static conditions. After 24 hours, the culture broth was centrifuged, and the supernatant was analyzed by HPLC to measure the amounts of sodium glutamate, GABA, arginine, and ornithine. As a result, GABA was produced at a conversion rate of 98%, and ornithine was produced at a conversion rate of 98%. It was. When this culture supernatant was freeze-dried and pulverized, 64.3 g of a brown powder containing 41% by weight of GABA and 11% by weight of ornithine was obtained.

Example 2
To 1 L of GYP medium containing 1.0% by weight sodium glutamate and 3.0% by weight arginine, 10 mL of a cell suspension of Lactobacillus brevis UAS-6 that has been pre-cultured and is in a steady state is added. The culture was started at 30 ° C. under static conditions. After 24 hours, the culture broth was centrifuged, and the supernatant was analyzed by HPLC to measure the amounts of sodium glutamate, GABA, arginine, and ornithine. As a result, GABA was produced at a conversion rate of 99%, and ornithine was produced at a conversion rate of 97%. It was. When this culture supernatant was freeze-dried and pulverized, 54.9 g of a brown powder containing 9.8% by weight of GABA and 52% by weight of ornithine was obtained.

Example 3
To a 20% non-fat dry milk solution, 1% protease (manufactured by Amano Pharmaceutical Co., Ltd., protease A “Amano G”) was added to the protein and allowed to react at 50 ° C. for 24 hours. Bacterial cells of Lactobacillus brevis UAS-4 pre-cultured in 1 L of a medium containing 15%, skim milk 20%, fructose 0.5%, and sodium glutamate 0.05%. 10 mL of the suspension was added and culture was started at 30 ° C. and standing. After 36 hours, the free amino acid content in the fermented milk was measured. Table 1 shows the contents of glutamic acid, GABA, arginine and ornithine before and after fermentation.

When this fermented milk was freeze-dried, 210 g of powder was obtained. The GABA and ornithine contents of this powder were 0.27% and 0.13%, respectively.

Example 4
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 2.5%, and then 1 w / w% of yeast extract was added to 1 L of each raw material filtrate, followed by high-pressure steam sterilization. Thereafter, 0.5 w / w% sodium glutamate and 0.3 w / w% arginine were added, and 10 ml of a preculture of Lactobacillus brevis UAS-4 was inoculated. After fermentation at 30 ° C. for 24 hours, the mixture was sterilized by heating at 100 ° C. for 5 minutes, and suction filtration was performed using a filter paper to obtain a clear filtrate. The filtrate was lyophilized to powder and the GABA and ornithine contents were measured. Table 2 shows the contents of GABA and ornithine in the freeze-dried product obtained for each raw material.

As shown in the table, each raw material contained about 8-9% GABA and about 6-7% ornithine.

Example 5
100 mg of the powder obtained in Examples 1 and 2 was dissolved in 200 ml of oolong tea to obtain oolong tea containing GABA and ornithine. The powder dissolved rapidly and there was almost no change in taste and odor.

Example 6
The freeze-dried product of each plant extract fermented liquid obtained in Example 4 was mixed with medium strength flour so as to be 1 w / w% to prepare noodles of udon. When the udon noodles were cooked, they were good noodles with a slight sweetness and a refreshing scent.

Claims (8)

Gamma-aminobutyric acid characterized by inoculating a medium containing glutamic acid or / and salts thereof and arginine or / and salts thereof with lactic acid bacteria having both gamma-aminobutyric acid-producing ability and ornithine-producing ability and performing lactic acid fermentation And a method for producing a composition containing ornithine. A method for producing a food containing γ-aminobutyric acid and ornithine, wherein a lactic acid bacterium having both γ-aminobutyric acid producing ability and ornithine producing ability is inoculated to a food material and lactic acid fermented. The method for producing a food containing γ-aminobutyric acid and ornithine according to claim 2, wherein glutamic acid or / and a salt thereof is further contained in the food material. The method for producing a food containing γ-aminobutyric acid and ornithine according to claim 2 or 3, wherein the food material further contains arginine or / and a salt thereof. The method for producing a food containing γ-aminobutyric acid and ornithine according to any one of claims 2 to 4, wherein the food material is pretreated with a proteolytic enzyme. The composition containing γ-aminobutyric acid and ornithine according to any one of claims 1 to 6, wherein the lactic acid bacterium having both γ-aminobutyric acid producing ability and ornithine producing ability is a lactic acid bacterium belonging to Lactobacillus brevis. Manufacturing method of food or food. The method for producing a food containing γ-aminobutyric acid and ornithine according to any one of claims 2 to 6, wherein the food material is obtained from asparagus. A composition or food containing γ-amino acid and ornithine produced by the production method according to claim 1.