JP2003180389A - METHOD FOR PRODUCING gamma-AMINOBUTYRIC ACID - Google Patents
METHOD FOR PRODUCING gamma-AMINOBUTYRIC ACIDInfo
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- JP2003180389A JP2003180389A JP2001382702A JP2001382702A JP2003180389A JP 2003180389 A JP2003180389 A JP 2003180389A JP 2001382702 A JP2001382702 A JP 2001382702A JP 2001382702 A JP2001382702 A JP 2001382702A JP 2003180389 A JP2003180389 A JP 2003180389A
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- JP
- Japan
- Prior art keywords
- aminobutyric acid
- acid
- liquid sugar
- lactic acid
- aminobutyric
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
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- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
Description
【０００１】[0001]
【発明の属する技術分野】本発明は、食品添加用のγ−
アミノ酪酸の生成方法に関する。TECHNICAL FIELD The present invention relates to γ-for food additives.
The present invention relates to a method for producing aminobutyric acid.
【０００２】[0002]
【発明が解決しようとする課題】γ−アミノ酪酸（ＧＡ
ＢＡ）は、血圧上昇抑制作用を有するアミノ酸であり、
高血圧の治療・予防に有効な機能性食品素材として近年
注目を浴びている。γ−アミノ酪酸は、微生物に広く分
布するグルタミン酸デカルボキシラーゼ（ＧＡＤ）の作
用によりグルタミン酸が脱炭酸されて生成する。SUMMARY OF THE INVENTION γ-Aminobutyric acid (GA
BA) is an amino acid having a blood pressure elevation suppressing effect,
In recent years, it has attracted attention as a functional food material that is effective in treating and preventing hypertension. γ-Aminobutyric acid is produced by decarboxylation of glutamic acid by the action of glutamate decarboxylase (GAD) widely distributed in microorganisms.
【０００３】微生物によるγ−アミノ酪酸の生成方法と
しては、例えば特開平６−４５１４１号公報に開示され
るような、乳酸菌を含有する醸造諸味を添加した魚醤油
諸味を発酵させる方法、特開平９−２３８６５０号公報
に開示されているような、グルタミン酸に酵母を作用さ
せる方法、特開平１０−１６５１９１号公報に開示され
ているような、麹菌アスペリギルスを培養する方法、特
開平１０−２９５３９４号公報に開示されているよう
な、発酵乳製品中の乳酸菌の脱炭素酵素作用による方
法、特開２０００−３０８４５７号公報に開示されてい
るような、脱脂乳とトマト果汁とを含む培地に２種の乳
酸菌を接種して混合培養し脱脂乳を乳酸発酵させる方
法、さらには、特開２００１−１２０１７９号公報に開
示されているような、乳製品をプロテアーゼ処理してか
ら乳酸菌等を接種して培養する方法などが知られてい
る。As a method for producing γ-aminobutyric acid by microorganisms, for example, a method of fermenting fish soy sauce moromi mash containing lactic acid bacteria-containing brewing moromi mash, as disclosed in Japanese Patent Application Laid-Open No. 6-45141, is disclosed in Japanese Unexamined Patent Application Publication No. Hei 9 (1998) -96952. As disclosed in JP-A-238650, a method of allowing yeast to act on glutamic acid, a method of culturing Aspergillus oryzae as disclosed in JP-A-10-165191, and JP-A-10-295394. A method by decarboxylase action of lactic acid bacteria in fermented milk products as disclosed, and two types of lactic acid bacteria in a medium containing skim milk and tomato juice as disclosed in JP 2000-308457 A Lactic acid fermentation of skim milk by inoculating and culturing mixed milk, and further milk as disclosed in JP 2001-120179 A And a method for culturing by inoculating the lactic acid bacteria the goods after protease treatment is known.
【０００４】しかしながら、これらの方法によって得ら
れるγ−アミノ酪酸の生成量は決して多いとはいえな
い。However, the amount of γ-aminobutyric acid produced by these methods is not very high.
【０００５】なお、工業的に生産されたグルタミン酸デ
カルボキシラーゼを用いれば、効率良くγ−アミノ酪酸
を生成できることが知られているが、グルタミン酸デカ
ルボキシラーゼは高価であり、反応条件も複雑なため、
実用化が難しい。It is known that γ-aminobutyric acid can be efficiently produced by using industrially produced glutamic acid decarboxylase, but since glutamic acid decarboxylase is expensive and the reaction conditions are complicated,
Practical application is difficult.
【０００６】本発明かかる課題に鑑みてなされたもので
あり、微生物が有するグルタミン酸デカルボキシラーゼ
の作用を利用して、簡単に効率良くグルタミン酸を脱炭
酸してγ−アミノ酪酸を生成することが可能な、γ−ア
ミノ酪酸の生成方法を提供することを目的とする。The present invention has been made in view of the above problems, and it is possible to easily and efficiently decarboxylate glutamic acid to produce γ-aminobutyric acid by utilizing the action of glutamic acid decarboxylase possessed by microorganisms. , Γ-aminobutyric acid is produced.
【０００７】[0007]
【課題を解決するための手段】上記目的に鑑み鋭意研究
の結果、本発明者は、微生物が有するグルタミン酸デカ
ルボキシラーゼの作用を利用して、簡単に効率良くグル
タミン酸を脱炭酸してγ−アミノ酪酸を生成することが
できる方法について種々検討したところ、液糖に鰹節と
蛋白質分解酵素とを加えて処理し、グルタミン酸又はそ
の塩類を加えて調合した後、乳酸菌を植菌して培養すれ
ばよいことを見出し、本発明に想到したものである。Means for Solving the Problems As a result of earnest research in view of the above object, the present inventor utilized the action of glutamic acid decarboxylase possessed by microorganisms to easily and efficiently decarboxylate glutamic acid to give γ-aminobutyric acid. As a result of various studies on a method capable of producing lactic acid, liquid sugar was treated with bonito and a proteolytic enzyme, treated with glutamic acid or a salt thereof, mixed, and then inoculated with lactic acid bacteria and cultured. The present invention has been made and the present invention has been conceived.
【０００８】本発明の請求項１記載のγ−アミノ酪酸の
生成方法は、液糖に鰹節と蛋白質分解酵素とを加えて処
理し、グルタミン酸又はその塩類を加えて調合した後、
乳酸菌を植菌して培養する方法である。In the method for producing γ-aminobutyric acid according to claim 1 of the present invention, liquid sugar is treated with bonito knot and proteolytic enzyme, treated, and then glutamic acid or a salt thereof is added to prepare the mixture.
This is a method in which lactic acid bacteria are inoculated and cultured.
【０００９】また、本発明の請求項２記載のγ−アミノ
酪酸の生成方法は、前記請求項１において、前記乳酸菌
がラクトバチルス・ブレビスである方法である。A method for producing γ-aminobutyric acid according to claim 2 of the present invention is the method according to claim 1, wherein the lactic acid bacterium is Lactobacillus brevis.
【００１０】さらに、本発明の請求項３記載のγ−アミ
ノ酪酸の生成方法は、前記請求項１又は２において、前
記液糖が米液糖である方法である。Further, a method for producing γ-aminobutyric acid according to claim 3 of the present invention is the method according to claim 1 or 2, wherein the liquid sugar is rice liquid sugar.
【００１１】[0011]
【発明の実施形態】以下、本発明のγ−アミノ酪酸の生
成方法について詳細に説明する。BEST MODE FOR CARRYING OUT THE INVENTION The method for producing γ-aminobutyric acid of the present invention will be described in detail below.
【００１２】本発明において液糖とは、液状ブドウ糖、
異性化液糖（果糖ブドウ糖液糖、ブドウ糖果糖液糖）、
砂糖混合異性化液糖、水あめ、コーンシロップ等のこと
である。この液糖は乳酸菌を培養する際の栄養源となる
ものであり、乳酸菌の栄養源となり得る液状の糖であれ
ば上記に限らない。In the present invention, liquid sugar means liquid glucose,
Isomerized liquid sugar (fructose glucose liquid sugar, glucose fructose liquid sugar),
It is sugar mixed isomerized liquid sugar, starch syrup, corn syrup and the like. This liquid sugar is a nutrient source when culturing lactic acid bacteria, and is not limited to the above as long as it is a liquid sugar that can be a nutrient source for lactic acid bacteria.
【００１３】また、本発明において米液糖とは、米ので
んぷんに液化酵素（アルファアミラーゼ）、糖化酵素
（グルコアミラーゼ）等を作用させて液状ブドウ糖とし
たもの、またはこのブドウ糖にさらに異性化酵素（グル
コースイソメラーゼ）を作用させて異性化液糖（果糖ブ
ドウ糖液糖、ブドウ糖果糖液糖）としたもの等のことで
あり、米を主原料としたものである。米液糖には、米に
含まれる栄養分であるビタミンＥ、ビタミンＢ1、ビタ
ミンＢ2、ナイアシン等のビタミン、カルシウム、鉄、
リン等のミネラルが含まれている。In the present invention, the rice liquor is a liquid glucose prepared by reacting liquefying enzyme (alpha amylase), saccharifying enzyme (glucoamylase) or the like on rice starch, or isomerase to this glucose. (Glucose isomerase) to produce isomerized liquid sugar (fructose glucose liquid sugar, glucose fructose liquid sugar) and the like, which is mainly rice. The liquid sugar of rice includes nutrients contained in rice such as vitamin E, vitamin B 1 , vitamin B 2 , and niacin, calcium, iron,
Contains minerals such as phosphorus.
【００１４】また、本発明においてグルタミン酸又はそ
の塩類としては、グルタミン酸、グルタミン酸ナトリウ
ム等が挙げられるが、液糖への溶解性を考慮すると、グ
ルタミン酸ナトリウムが最も好ましい。このグルタミン
酸又はその塩類は、γ−アミノ酪酸の原料となる。In the present invention, examples of glutamic acid or salts thereof include glutamic acid and sodium glutamate, but sodium glutamate is most preferable in view of solubility in liquid sugar. This glutamic acid or its salts serve as a raw material for γ-aminobutyric acid.
【００１５】また、本発明において蛋白質分解酵素（プ
ロテアーゼ）としては、微生物、植物、動物由来のもの
のいずれでもよい。例えば、プロテアーゼＭアマノ（天
野製薬製）、パンチターゼＮＰ−２（ヤクルト薬品工業
製）、パパイン（アサヒビール食品製）等が挙げられ
る。In the present invention, the proteolytic enzyme (protease) may be derived from microorganisms, plants or animals. For example, Protease M Amano (manufactured by Amano Pharmaceutical Co., Ltd.), Punchase NP-2 (manufactured by Yakult Chemical Industry), papain (manufactured by Asahi Breweries Foods) and the like can be mentioned.
【００１６】また、本発明において乳酸菌としては、グ
ルタミン酸デカルボキシラーゼの作用を有し、食品に添
加できるものであればよく、例えば、ラクトバチルス・
ブルガリクス、ラクトバチルス・ヘルベティクス、ラク
トバチルス・アシドフィルス、ラクトバチルス・カゼ
イ、ラクトバチルス・プランタルム、ラクトバチルス・
ファーメンタム、ラクトバチルス・ブレビス等のラクト
バチルス属、ビフィドバクテリウム・ブレーベ、ビフィ
ドバクテリウム・ビフィダム、ビフィドバクテリウム・
インファンティス、ビフィドバクテリウム・ロンガム、
ビフィドバクテリウム・アドレッセンティス、ビフィド
バクテリウム・サーモフィラム、ビフィドバクテリウム
・シュードロンガム等のビフィドバクテリウム属、スト
レプトコッカス・ラクティス、ストレプトコッカス・ク
レモリス、ストレプトコッカス・サーモフィルス、スト
レプトコッカス・フェカーリス等のストレプトコッカス
属、ペディオコッカス・セレビシェ、ペディオコッカス
・ハロフィルス、ペディオコッカス・パーバルス等のペ
ディオコッカス属、ロイコノストック・メゼントロイデ
ス、ロイコノストック・シトロボラム等のロイコノスト
ック属等が挙げられる。Further, in the present invention, the lactic acid bacterium may be any lactic acid bacterium as long as it has a function of glutamate decarboxylase and can be added to foods.
Bulgarix, Lactobacillus helvetics, Lactobacillus acidophilus, Lactobacillus casei, Lactobacillus plantarum, Lactobacillus
Lactobacillus such as fermentum, Lactobacillus brevis, Bifidobacterium breve, Bifidobacterium bifidum, Bifidobacterium
Infantis, Bifidobacterium longum,
Bifidobacterium genus such as Bifidobacterium adressetis, Bifidobacterium thermophilum, Bifidobacterium pseudolongum, Streptococcus lactis, Streptococcus cremoris, Streptococcus thermophilus, Streptococcus faecalis, etc. Examples include genus Pediococcus such as Streptococcus, Pediococcus cerevisiae, Pediococcus halophilus, Pediococcus pervals, Leuconostoc mesentroides, and Leuconostoc genus such as Leuconostoc citrovolum.
【００１７】つぎに、上述したような液糖、グルタミン
酸又はその塩類、蛋白質分解酵素、乳酸菌を用いた本発
明のγ−アミノ酪酸の生成方法について説明する。Next, a method for producing γ-aminobutyric acid of the present invention using the above-mentioned liquid sugar, glutamic acid or salts thereof, proteolytic enzyme and lactic acid bacterium will be described.
【００１８】まず、液糖に鰹節を加え、均一に混合して
調合する。この鰹節の添加量は、液糖１００重量部に対
して１〜２０重量部、好ましくは３〜１０重量部であ
る。鰹節が１重量部未満ではγ−アミノ酪酸の生成量が
少なくなり、一方、２０重量部を超えても鰹節の添加量
増加分の効果が期待できないばかりか、使用済の鰹節を
取り除く後処理の負担が増加するため好ましくない。な
お、鰹節を粉末状にすることによって、γ−アミノ酪酸
の生成効率が高まるとともに、濾過による使用済の鰹節
の除去が容易となる。First, bonito flakes are added to liquid sugar and mixed uniformly to prepare. The amount of bonito flakes added is 1 to 20 parts by weight, preferably 3 to 10 parts by weight, based on 100 parts by weight of liquid sugar. When the amount of bonito is less than 1 part by weight, the amount of γ-aminobutyric acid produced is small. On the other hand, when the amount of bonito is more than 20 parts by weight, the effect of the increased amount of bonito is not expected, and the post-treatment for removing used bonito is required. This is not preferable because it increases the burden. By making the bonito flakes into powder, the production efficiency of γ-aminobutyric acid is increased, and the used bonito flakes can be easily removed by filtration.
【００１９】そして、蛋白質分解酵素を添加し、蛋白質
分解酵素の活性が高い温度である３０〜７０℃、好まし
くは５０〜６０℃で、攪拌しながら１〜１０時間、好ま
しくは２〜５時間保温する。ここでは、鰹節の蛋白質を
ペプチドやアミノ酸へ分解する。使用する酵素量は、力
価として５，５００Ｕ／ｇ以上のものを、酵素蛋白質の
総量として、鰹節１００重量部に対し０．１〜０．４重
量部とするのが、γ−アミノ酪酸の生成量、蛋白質分解
酵素のコストの面から好ましい。Then, a proteolytic enzyme is added, and the temperature is maintained at 30 to 70 ° C., preferably 50 to 60 ° C., where the activity of the proteolytic enzyme is high, while stirring and for 1 to 10 hours, preferably 2 to 5 hours. To do. Here, the protein of bonito is decomposed into peptides and amino acids. The amount of enzyme used is 5,500 U / g or more as a titer, and the total amount of enzyme protein is 0.1 to 0.4 parts by weight with respect to 100 parts by weight of bonito flakes. It is preferable in terms of the production amount and the cost of the protease.
【００２０】つぎに、この反応溶液にグルタミン酸又は
その塩類を加えて調合する。このグルタミン酸又はその
塩類の添加量は、グルタミン酸ナトリウム換算で、液糖
１００重量部に対して５〜２０重量部、好ましくは６〜
１０重量部である。グルタミン酸又はその塩類が５重量
部未満ではγ−アミノ酪酸を多量に生成できず、一方、
２０重量部を超えると未反応のグルタミン酸又はその塩
類が多量に残存するため好ましくない。そして、８０〜
１００℃で３〜６０分間、好ましくは８０〜８５℃で３
０〜６０分間加熱し、蛋白質分解酵素を失活させるとと
もに、殺菌する。Next, glutamic acid or its salt is added to this reaction solution to prepare a mixture. The amount of glutamic acid or its salt added is 5 to 20 parts by weight, preferably 6 to 10 parts by weight, based on 100 parts by weight of liquid sugar, in terms of sodium glutamate.
10 parts by weight. If less than 5 parts by weight of glutamic acid or a salt thereof cannot produce a large amount of γ-aminobutyric acid,
If it exceeds 20 parts by weight, a large amount of unreacted glutamic acid or a salt thereof remains, which is not preferable. And 80 ~
3 to 60 minutes at 100 ° C., preferably 3 to 80 to 85 ° C.
Heat for 0 to 60 minutes to inactivate proteolytic enzymes and sterilize.
【００２１】その後、上記で得た反応溶液を２０〜４０
℃に冷却し、この反応溶液に乳酸菌を加える。乳酸菌と
しては、グルタミン酸デカルボキシラーゼの作用の強い
乳酸菌である、ラクトバチルス・ブレビスが最も好まし
い。乳酸菌の添加量は、反応溶液１００重量部に対し
て、０．５〜５重量部、好ましくは０．５〜２重量部と
する。乳酸菌が０．５重量部未満ではγ−アミノ酪酸の
生成に時間を要し、一方、５重量部を超えても乳酸菌の
添加量増加分の効果が期待できないため好ましくない。
そして、２０〜４０℃で１００〜２００時間程度、静置
して培養する。上記培養時の温度と時間は一例であり、
乳酸菌の種類等に応じて適宜調節するとよい。ここで培
養される乳酸菌が液糖を栄養分としてグルタミン酸又は
その塩類を脱炭酸することによって、γ−アミノ酪酸が
生成する。反応終了後、鰹節を濾過により除去すること
によって、γ−アミノ酪酸を多量に含んだ液糖が得られ
る。Thereafter, the reaction solution obtained above is added to 20-40
Cool to 0 ° C. and add lactic acid bacteria to the reaction solution. The most preferred lactic acid bacterium is Lactobacillus brevis, which is a lactic acid bacterium having a strong action of glutamate decarboxylase. The amount of lactic acid bacteria added is 0.5 to 5 parts by weight, preferably 0.5 to 2 parts by weight, based on 100 parts by weight of the reaction solution. If the amount of lactic acid bacteria is less than 0.5 parts by weight, it takes time to produce γ-aminobutyric acid, while if it exceeds 5 parts by weight, the effect of increasing the amount of added lactic acid bacteria cannot be expected, which is not preferable.
And it culture | cultivates by leaving still at 20-40 degreeC for about 100-200 hours. The temperature and time during the above culture are examples,
It may be appropriately adjusted depending on the type of lactic acid bacteria. The lactic acid bacterium cultured here decarboxylates glutamic acid or a salt thereof using liquid sugar as a nutrient, to produce γ-aminobutyric acid. After completion of the reaction, the bonito is removed by filtration to obtain a liquid sugar containing a large amount of γ-aminobutyric acid.
【００２２】以上詳述したとおり、本発明のγ−アミノ
酪酸の生成方法は、液糖に鰹節と蛋白質分解酵素とを加
えて処理し、グルタミン酸又はその塩類を加えて調合し
た後、乳酸菌を植菌して培養するものであるので、乳酸
菌が有するグルタミン酸デカルボキシラーゼの作用を利
用して、簡単に効率良くグルタミン酸を脱炭酸してγ−
アミノ酪酸を生成することができる。特に、乳酸菌とし
てラクトバチルス・ブレビスを選択することにより、高
効率でγ−アミノ酪酸を生成することができる。また、
液糖として米液糖を選択することによって、γ−アミノ
酪酸のみならず、米独自の栄養素をも含んだ液糖を得る
ことができる。As described above in detail, in the method for producing γ-aminobutyric acid of the present invention, liquid sugar is treated with bonito and proteolytic enzymes, treated with glutamic acid or its salts, and then mixed with lactic acid bacteria. Since it is cultivated as a bacterium, the action of glutamic acid decarboxylase possessed by lactic acid bacteria is utilized to easily and efficiently decarboxylate glutamic acid to produce γ-
Aminobutyric acid can be produced. In particular, by selecting Lactobacillus brevis as the lactic acid bacterium, γ-aminobutyric acid can be produced with high efficiency. Also,
By selecting rice liquid sugar as the liquid sugar, it is possible to obtain liquid sugar containing not only γ-aminobutyric acid but also rice-specific nutrients.
【００２３】さらに、本発明のγ−アミノ酪酸の生成方
法によって得られたγ−アミノ酪酸を多量に含んだ液糖
を甘味料として食品に添加すれば、簡単にγ−アミノ酪
酸を多量に含んだ食品、例えば、ヨーグルト、飲料、菓
子等を提供することができる。また、本発明のγ−アミ
ノ酪酸の生成方法によれば、高効率でγ−アミノ酪酸を
生成し、残存するグルタミン酸又はその塩類の量が少な
いため、グルタミン酸又はその塩類が上記食品の風味に
悪影響を及ぼすこともない。Furthermore, if the liquid sugar containing a large amount of γ-aminobutyric acid obtained by the method for producing γ-aminobutyric acid of the present invention is added as a sweetener to foods, a large amount of γ-aminobutyric acid can be easily contained. Food products such as yogurt, beverages, confectionery and the like can be provided. Further, according to the method for producing γ-aminobutyric acid of the present invention, γ-aminobutyric acid is produced with high efficiency, and the amount of residual glutamic acid or its salts is small, so glutamic acid or its salts adversely affects the flavor of the food. It does not affect.
【００２４】[0024]
【実施例】以下の具体的実施例により、本発明をさらに
詳細に説明する。The present invention will be described in more detail with reference to the following specific examples.
【００２５】実施例
米に液化酵素、糖化酵素を作用させて液状ブドウ糖とし
た米液糖３００ｇに粉末状の鰹節３０ｇを加え、均一に
混合して調合した。そして、蛋白質分解酵素プロテアー
ゼＭアマノを２５８０Ｕ添加し、ｐＨ無調整で攪拌しな
がら５５℃で３時間保温して反応させた。つぎに、この
反応溶液にグルタミン酸ナトリウム２０ｇを加えて調合
した。そして、８０℃で３０分間加熱して蛋白質分解酵
素を失活させるとともに、殺菌した。 Example To 30 g of powdered bonito flakes was added to 300 g of rice syrup which was made into liquid glucose by reacting liquefying enzyme and saccharifying enzyme with rice, and mixed uniformly to prepare. Then, 2580 U of proteolytic enzyme Protease M Amano was added, and the mixture was allowed to react at 55 ° C. for 3 hours while stirring without pH adjustment. Next, 20 g of sodium glutamate was added to this reaction solution to prepare a mixture. Then, it was heated at 80 ° C. for 30 minutes to inactivate the proteolytic enzyme and sterilized.
【００２６】その後、上記で得た反応溶液を３２℃に冷
却し、ラクトバチルス・ブレビスを２ｇ加えて３２℃で
４８時間、静置して培養し、その後さらに２４℃で１２
０時間培養した。培養終了後、鰹節を濾過により除去し
培養液２５０ｇを得た。高速アミノ酸分析計Ｌ−８８０
０（日立製）を用いて分析したところ、培養液１００ｇ
中のγ−アミノ酪酸は４，２０８．７７ｍｇであった。
また、残存したグルタミン酸ナトリウムは培養液１００
ｇあたり２５９ｍｇであった。グルタミン酸ナトリウム
調合時の調合液１００ｇ中のグルタミン酸ナトリウム
は、５，７１４ｍｇであったので、濃度を基準にしたグ
ルタミン酸ナトリウムの残存率は、僅か４．５％であっ
た。Then, the reaction solution obtained above was cooled to 32 ° C., 2 g of Lactobacillus brevis was added, and the mixture was allowed to stand at 32 ° C. for 48 hours for culturing, and then at 24 ° C. for 12 hours.
It was cultured for 0 hours. After completion of the culture, the bonito was removed by filtration to obtain 250 g of a culture solution. High Speed Amino Acid Analyzer L-880
When analyzed using 0 (manufactured by Hitachi), 100 g of culture solution
The amount of γ-aminobutyric acid therein was 4,208.77 mg.
In addition, the remaining sodium glutamate is 100
It was 259 mg per gram. Since sodium glutamate in the preparation liquid 100 g at the time of preparation of sodium glutamate was 5,714 mg, the residual rate of sodium glutamate based on the concentration was only 4.5%.
【００２７】さらに、上記と全く同様の条件で反応、培
養を行った結果、培養液１００ｇ中のγ−アミノ酪酸は
４，１３４．２９ｍｇであり、再現性が確認できた。こ
れらの結果を表１に示す。Further, as a result of the reaction and culturing under exactly the same conditions as above, γ-aminobutyric acid in 100 g of the culture broth was 4,134.29 mg, confirming reproducibility. The results are shown in Table 1.
【００２８】比較例１〜３
比較のために、実施例で用いた鰹節の代わりに、同量の
さば節、煮干し、昆布を添加物として加え、その他の条
件は実施例と全く同様の条件で反応、培養を行った結果
を表１に示す。なお、昆布を用いた場合のみ、蛋白質分
解酵素を用いた処理を行わなかった。 Comparative Examples 1 to 3 For comparison, instead of the bonito flakes used in Examples, the same amount of mackerel, dried sardines, and kelp were added as additives, and other conditions were exactly the same as those of Examples. Table 1 shows the results of the reaction and culturing carried out in. The treatment with proteolytic enzyme was not carried out only when kelp was used.
【００２９】[0029]
【表１】 [Table 1]
【００３０】表１から明らかなとおり、本実施例のγ−
アミノ酪酸の生成方法によれば、鰹節を用いることによ
り非常に効果的にγ−アミノ酪酸を生成できることがわ
かった。これに対し、鰹節の代わりにさば節、煮干し、
昆布を用いた比較例１〜３では、γ−アミノ酪酸の生成
量が非常に少なかった。また、蛋白質分解酵素を用いた
処理を行わなかった比較例３ではγ−アミノ酪酸の生成
は確認できなかった。以上の実験結果から、液糖に鰹節
と蛋白質分解酵素とを加えて処理し、グルタミン酸又は
その塩類を加えて調合した後、乳酸菌を植菌して培養す
る本発明の方法によりγ−アミノ酪酸を簡単に効率良く
生成できることがわかった。As is clear from Table 1, γ-of this example
According to the method for producing aminobutyric acid, it was found that γ-aminobutyric acid can be produced very effectively by using bonito flakes. On the other hand, instead of bonito flakes, mackerel, dried sardines,
In Comparative Examples 1 to 3 using kelp, the amount of γ-aminobutyric acid produced was very small. Further, in Comparative Example 3 in which the treatment with the proteolytic enzyme was not performed, the production of γ-aminobutyric acid could not be confirmed. From the above experimental results, liquid sugar was treated by adding bonito and proteolytic enzymes, and after adding glutamic acid or a salt thereof to prepare γ-aminobutyric acid by the method of the present invention inoculating and culturing lactic acid bacteria. It turns out that it can be generated easily and efficiently.
【００３１】比較例４〜６
上記実施例で用いた乳酸菌ラクトバチルス・ブレビスの
代わりに、同量の乳酸菌ラクトバチルス・プランタル
ム、ロイコノストック・メゼントロイデス、ペディオコ
ッカス・パーバルスを加え、その他の条件は実施例と全
く同様の条件で反応、培養を行った結果を表２に示す。 Comparative Examples 4 to 6 Instead of the lactic acid bacterium Lactobacillus brevis used in the above Examples, the same amounts of the lactic acid bacterium Lactobacillus plantarum, Leuconostoc mesentroides and Pediococcus pervals were added, and the other conditions were Table 2 shows the results of the reaction and culturing performed under exactly the same conditions as in the examples.
【００３２】[0032]
【表２】 [Table 2]
【００３３】表２から明らかなとおり、ラクトバチルス
・プランタルム、ロイコノストック・メゼントロイデ
ス、ペディオコッカス・パーバルスを用いた場合には、
γ−アミノ酪酸の生成は確認されなかった。以上の実験
結果から、乳酸菌としてはラクトバチルス・ブレビスが
好ましく、乳酸菌としてラクトバチルス・ブレビスを用
いることによって、高効率でγ−アミノ酪酸を生成でき
ることがわかった。As is clear from Table 2, when Lactobacillus plantarum, Leuconostoc mesentroides and Pediococcus pervals were used,
No production of γ-aminobutyric acid was confirmed. From the above experimental results, it was found that Lactobacillus brevis is preferable as the lactic acid bacterium, and by using Lactobacillus brevis as the lactic acid bacterium, γ-aminobutyric acid can be produced with high efficiency.
【００３４】[0034]
【発明の効果】本発明の請求項１記載のγ−アミノ酪酸
の生成方法は、液糖に鰹節と蛋白質分解酵素とを加えて
処理し、グルタミン酸又はその塩類を加えて調合した
後、乳酸菌を植菌して培養するものであるので、乳酸菌
が有するグルタミン酸デカルボキシラーゼの作用を利用
して、簡単に効率良くグルタミン酸を脱炭酸してγ−ア
ミノ酪酸を生成することができる。The method for producing γ-aminobutyric acid according to claim 1 of the present invention is a method in which liquid sugar is treated with bonito and a proteolytic enzyme, treated with glutamic acid or a salt thereof, and then mixed with lactic acid bacteria. Since the cells are inoculated and cultured, the action of glutamate decarboxylase possessed by lactic acid bacteria can be utilized to easily and efficiently decarboxylate glutamic acid to produce γ-aminobutyric acid.
【００３５】また、請求項２記載のγ−アミノ酪酸の生
成方法は、前記請求項１において、乳酸菌がラクトバチ
ルス・ブレビスである方法であるので、高効率でγ−ア
ミノ酪酸を生成することができる。The method for producing γ-aminobutyric acid according to claim 2 is the method according to claim 1, in which the lactic acid bacterium is Lactobacillus brevis, so that γ-aminobutyric acid can be produced with high efficiency. it can.
【００３６】さらに、本発明の請求項３記載のγ−アミ
ノ酪酸の生成方法は、前記請求項１又は２において、前
記液糖が米液糖である方法であるので、γ−アミノ酪酸
のみならず、米独自の栄養素をも含んだ液糖を得ること
ができる。Furthermore, the method for producing γ-aminobutyric acid according to claim 3 of the present invention is the method according to claim 1 or 2, wherein the liquid sugar is rice liquid sugar. Instead, liquid sugar containing nutrients unique to rice can be obtained.
Claims (3)
処理し、グルタミン酸又はその塩類を加えて調合した
後、乳酸菌を植菌して培養することを特徴とするγ−ア
ミノ酪酸の生成方法。1. Production of γ-aminobutyric acid, characterized in that liquid sugar is treated with bonito and proteolytic enzymes, treated with glutamic acid or a salt thereof, and mixed, and then lactic acid bacteria are inoculated and cultured. Method.
であることを特徴とする請求項１記載のγ−アミノ酪酸
の生成方法。2. The method for producing γ-aminobutyric acid according to claim 1, wherein the lactic acid bacterium is Lactobacillus brevis.
る請求項１又は２に記載のγ−アミノ酪酸の生成方法。3. The method for producing γ-aminobutyric acid according to claim 1, wherein the liquid sugar is rice liquid sugar.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR100687599B1 (en) * | 2004-11-10 | 2007-02-27 | (주)바이오벤 | Method for Preparing ???? Using By-Products from Rice Polishing |
| WO2007097374A1 (en) * | 2006-02-21 | 2007-08-30 | Kikkoman Corporation | Lactic acid bacterium capable of producing ϝ-aminobutyric acid |
| KR100857215B1 (en) | 2007-04-09 | 2008-09-05 | 주식회사 엠에이치투 바이오케미칼 | Method for preparing highly pure gamma;-amino butyric acid using enzymic reaction |
| CN111000175A (en) * | 2019-12-17 | 2020-04-14 | 四川东坡中国泡菜产业技术研究院 | Making method of low-putrescine healthy pickled vegetables |
-
2001
- 2001-12-17 JP JP2001382702A patent/JP3849045B2/en not_active Expired - Lifetime
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR100687599B1 (en) * | 2004-11-10 | 2007-02-27 | (주)바이오벤 | Method for Preparing ???? Using By-Products from Rice Polishing |
| WO2007097374A1 (en) * | 2006-02-21 | 2007-08-30 | Kikkoman Corporation | Lactic acid bacterium capable of producing ϝ-aminobutyric acid |
| US8202710B2 (en) | 2006-02-21 | 2012-06-19 | Kikkoman Corporation | Lactic acid bacterium capable of producing Y-aminobutyric acid |
| KR100857215B1 (en) | 2007-04-09 | 2008-09-05 | 주식회사 엠에이치투 바이오케미칼 | Method for preparing highly pure gamma;-amino butyric acid using enzymic reaction |
| CN111000175A (en) * | 2019-12-17 | 2020-04-14 | 四川东坡中国泡菜产业技术研究院 | Making method of low-putrescine healthy pickled vegetables |
| CN111000175B (en) * | 2019-12-17 | 2023-05-05 | 四川东坡中国泡菜产业技术研究院 | Method for preparing low-putrescine healthy pickle |
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| Publication number | Publication date |
|---|---|
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