JPS62275688A - Production of pyruvic acid by fermentation - Google Patents

Production of pyruvic acid by fermentation

Info

Publication number
JPS62275688A
JPS62275688A JP27531086A JP27531086A JPS62275688A JP S62275688 A JPS62275688 A JP S62275688A JP 27531086 A JP27531086 A JP 27531086A JP 27531086 A JP27531086 A JP 27531086A JP S62275688 A JPS62275688 A JP S62275688A
Authority
JP
Japan
Prior art keywords
pyruvic acid
torulopsis
producing
pyruvate
fermentation
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.)
Granted
Application number
JP27531086A
Other languages
Japanese (ja)
Other versions
JPH0358275B2 (en
Inventor
Toru Yonehara
徹 米原
Kyosuke Yomoto
四本 喬介
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Toray Industries Inc
Original Assignee
Toray Industries Inc
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Toray Industries Inc filed Critical Toray Industries Inc
Publication of JPS62275688A publication Critical patent/JPS62275688A/en
Publication of JPH0358275B2 publication Critical patent/JPH0358275B2/ja
Granted legal-status Critical Current

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  • Preparation Of Compounds By Using Micro-Organisms (AREA)

Abstract

PURPOSE:To produce pyruvic acid in high yield with little formation of by- product, by culturing a thiamine-requiring pyruvic acid-producing strain belonging to Torulopsis genus. CONSTITUTION:A pyruvic acid-producing strain belonging to Torulopsis genus and requiring thiamine for growth and preferably further vitamins such as biotin for growth, e.g. Torulopsis glabrata (IFO 0005), Torulopsis methanolovescens (ATCC 26176), etc., is cultured in a medium containing thiamine and, preferably further, vitamins such as biotin and pyruvic acid is separated from the cultured product.

Description

【発明の詳細な説明】 3、発明の詳細な説明 〈産業上の利用分野〉 本発明は発酵法によるピルビン酸の製造法に関するんも
のである。
[Detailed Description of the Invention] 3. Detailed Description of the Invention (Field of Industrial Application) The present invention relates to a method for producing pyruvic acid by a fermentation method.

ピルビン酸は生体代謝の重要な中間体であり、各硬固・
、@薬などの有効な合成原料で必るのみならず酸素法に
よるL−トップ1〜フアン、L−システィン、し−チロ
シンなどのアミノ酸合成の主要原料で必る。よって安価
に製造しくqれば、種々の合成原料として有用で必る。
Pyruvate is an important intermediate in biological metabolism, and is
It is essential not only as an effective raw material for the synthesis of drugs, but also as a main raw material for the synthesis of amino acids such as L-top 1-fan, L-cysteine, and tyrosine by the oxygen method. Therefore, if it can be produced at low cost, it will be useful as a raw material for various synthetics.

〈従来の技術〉 従来、発酵法によるピルビン酸の製造法としては、サツ
カロミセス属およびキャンディダ屈などの酵母菌や担子
菌類または特殊なバクテリアによる方法が知られている
(特公昭57−796号公報、特開昭57−’1594
92号公報等)。
<Prior art> Conventionally, as a method for producing pyruvic acid by fermentation, a method using yeasts such as Satucharomyces and Candida spp., basidiomycetes, or special bacteria is known (Japanese Patent Publication No. 57-796). , Japanese Patent Publication No. 57-'1594
Publication No. 92, etc.).

く本発明が解決しようとする問題点〉 しかしながら、かかる従来方法は副生物が多かったり、
または、収率・収量が十分でなかったりしたため工業的
に有利な方法とは言えなかった。
Problems to be Solved by the Present Invention> However, such conventional methods produce many by-products,
Alternatively, the yield and yield were insufficient, so it could not be said to be an industrially advantageous method.

く問題点を解決するための手段および作用〉したがって
本発明者らは上記問題点を解決することのできる、新規
な、発酵法によるピルビン酸生産菌を見出すことを目的
として鋭意研究したところ、トルロプシス居に属する酵
母菌が優れた効果を奏することを見出し本発明に到達し
た。
Means and Action for Solving the Problems> Therefore, the present inventors conducted intensive research with the aim of finding a new pyruvate-producing bacterium by fermentation that could solve the above problems, and found that Torulopsis The present invention was achieved by discovering that yeast bacteria belonging to the genus genus spp. have excellent effects.

すなわち、本発明の上記目的は、トルロプシス属に属す
るピルビン酸生産菌を培養することにより培地中にピル
ビン酸を生成蓄積させ、これを採取することにより、達
成されるのである。
That is, the above-mentioned object of the present invention is achieved by culturing pyruvate-producing bacteria belonging to the genus Torulopsis to produce and accumulate pyruvate in a medium, and then collecting the pyruvate.

トルロプシス属の酵母を用いた発酵法によるピルビン酸
の著量の蓄積はこれまで知られていなかった。
The accumulation of significant amounts of pyruvate by fermentation using yeast of the genus Torulopsis has not been known so far.

以下、本発明の構成を詳細に説明する。Hereinafter, the configuration of the present invention will be explained in detail.

本発明に厄いられるピルビン酸生産菌はピルビン酸生成
能を荷するトルロプシス属に属する酵母菌であればいか
なるものでおってもよいが、望ましくは各種のビタミン
を要求するもの、特にチアミン、ナイアシン、ピリドキ
シン、ビオチンなどを単独もしくは併せて要求するもの
が好適である。
The pyruvate-producing bacteria troubled by the present invention may be any yeast belonging to the genus Torulopsis that has the ability to produce pyruvate, but preferably those that require various vitamins, especially thiamin and niacin. , pyridoxine, biotin, etc. alone or in combination are suitable.

本発明で用いられるピルビン酸生産菌としては、例えば
、トルロプシス・グラブラータ(Torulopsis
  glabrata)()FO0005)(チアミン
、ビオチン、ピリドキシンおよびナイアシン要求)、ト
ルロプシス・メタノロベラセンス(Toru I op
s iS  methanoloVescens)(△
TCC26176>(チアミンおよびビオチン要求)な
どが挙げられる 本発明で用いられる培地は発酵に通常使用される炭素源
、窒素源、無機塩類、ビタミン類などを程よく含有する
ものであればよいが、炭素源としては、グルコースなど
の糖質、有機酸、エタノール、メタノールなどの使用酵
母菌が利用し得るものが使用される。窒素源として硫安
、硝安、塩安、尿素、ペプトン、肉エキス、味液、その
他の有機および無機窒素化合物が使用されるが、望まし
くはアミノ酸をバランスよく含む有機窒素化合物がよい
。無機塩類としてはリン酸カリウム、FiA酸マグネシ
ウム、鉄、マンガン、その他の無機塩類が用いられ、ざ
らに必要に応じてチアミン、ナイアシン、ピリドキシン
、ビオチンなどの要求ビタミン、またはこれらを含有す
る酵母エキス、コーン・スチープ・リカー、その他の天
然物を添加した培地を使用すればよい。
Examples of the pyruvate-producing bacteria used in the present invention include Torulopsis glabrata.
glabrata) ()FO0005) (requires thiamine, biotin, pyridoxine and niacin), Torulopsis methanoloverascens (Toru I op
s iS methanoloVescens) (△
TCC26176> (thiamin and biotin requirement), etc. The culture medium used in the present invention may contain moderate amounts of carbon sources, nitrogen sources, inorganic salts, vitamins, etc. normally used in fermentation. For example, carbohydrates such as glucose, organic acids, ethanol, methanol, etc., which can be used by the yeast used, are used. As a nitrogen source, ammonium sulfate, ammonium nitrate, ammonium chloride, urea, peptone, meat extract, flavor liquid, and other organic and inorganic nitrogen compounds are used, and organic nitrogen compounds containing amino acids in a well-balanced manner are preferable. Potassium phosphate, magnesium FiA, iron, manganese, and other inorganic salts are used as inorganic salts, and if necessary, required vitamins such as thiamin, niacin, pyridoxine, and biotin, or yeast extract containing these, A medium supplemented with corn steep liquor or other natural products may be used.

培養中はピルビン酸の生成蓄積にともない、ρ1」の低
下が起こるので炭酸カルシウム、苛性ソーダ、苛性カリ
などのアJレカリでpl」3〜7に調節することがピル
ビン酸生産には有効である。培養中の温度は22℃〜3
2°Cが適当でおる。培養終了後、系内に蓄積したピル
ビン酸は常法により、単離採取することができる。例え
ば、酸性エーテル抽出、フェニルヒドラゾン化して沈澱
単離する方法なども採用することができる。
During cultivation, as pyruvate is produced and accumulated, ρ1' decreases, so it is effective for pyruvic acid production to adjust pl' to 3 to 7 with an alimentary agent such as calcium carbonate, caustic soda, or caustic potash. The temperature during culturing is 22℃~3
2°C is suitable. After completion of the culture, pyruvate accumulated in the system can be isolated and collected by a conventional method. For example, methods such as acidic ether extraction and phenylhydrazonation followed by precipitation isolation can also be employed.

〈実施例〉 以下、実施例によって本発明を説明する。<Example> The present invention will be explained below with reference to Examples.

実施例において生成したピルビン酸の確認と定量は高速
液体クロマトグラフィーによる方法および乳酸脱水素酵
素による方法によって行った。以下の分析結果について
は上記両分析法ともよく合致してあり、同じ分析数値を
示した。
Confirmation and quantification of pyruvate produced in the Examples were performed by a method using high performance liquid chromatography and a method using lactate dehydrogenase. The following analytical results were in good agreement with both of the above analytical methods and showed the same analytical values.

実施例1 グルコース10%、ペプトン1%、K H2Pg/ff
、ピリドキシン塩酸塩100μg/fl、ニコチンa1
00tl’J/41.pl−15,0から’;する培地
30ml1を500 tni容振盛フラスコに分注滅菌
後、別に滅菌したC a C034%を添加し、トルロ
プシス・グラブラータ(IFO0005)を1白金耳植
菌した後、30’Cで91時間撮隔培養した。ビオチン
は添加しなくともペプトンに含まれる量で十分でおった
。培養後ピルビン酸を定量したところ40.7310.
で市り、残糖は223/αであった。副生物は液体クロ
マトグラフィーでは検出されなかった。この培養液1Q
を除菌後、上澄液に塩酸を加え112゜0とし、10.
のエチルエーテルで抽出し、次いで苛性ソーダでpHを
6.0に中和した後40℃で減圧濃縮し100d程度と
した。この濃縮液にエタノールを滴下させピルビン酸ソ
ーダ23.1(純度97%)を得た。
Example 1 Glucose 10%, peptone 1%, K H2Pg/ff
, pyridoxine hydrochloride 100μg/fl, nicotine a1
00tl'J/41. After sterilizing and dispensing 30 ml of medium from pl-15,0 to a 500 tni shaking flask, adding separately sterilized Ca C034% and inoculating one platinum loop of Torulopsis glabrata (IFO0005), Culture was carried out at 30'C for 91 hours at intervals. Even without adding biotin, the amount contained in peptone was sufficient. After culturing, pyruvate was quantified and found to be 40.7310.
The remaining sugar was 223/α. No by-products were detected by liquid chromatography. This culture solution 1Q
After sterilization, add hydrochloric acid to the supernatant to bring it to 112°0, and 10.
The extract was extracted with ethyl ether, then neutralized to pH 6.0 with caustic soda, and concentrated under reduced pressure at 40° C. to about 100 d. Ethanol was added dropwise to this concentrated solution to obtain sodium pyruvate 23.1 (purity 97%).

実施例2 実施例1において使用した培地中のグルコースを5%に
減らし、ピリドキシン塩酸塩およびニコチン酸をビオチ
ン0.5μg10.におきかえた以外は同様な培地を用
い、トルロプシス・グラブラータをトルロプシス・メタ
ノロベラセンス(ATCC26176)に変えて実施例
1と同様の方法で培養した。
Example 2 Glucose in the medium used in Example 1 was reduced to 5%, pyridoxine hydrochloride and nicotinic acid were added to biotin 0.5 μg 10. The culture was carried out in the same manner as in Example 1, except that the same medium was used, and Torulopsis glabrata was replaced with Torulopsis methanoloverascens (ATCC 26176).

72時間後、培養液中のピルビン酸を定量したところ2
0.5g/Qであり、残糖はなかった。 〈発明の効果
〉 本発明の方法は、発酵法によりピルビン酸を著量蓄積せ
しめることが可能であり、高収率、収量でピルビン酸を
得ることができるとともに、副生物がほとんど生成しな
いため、工業的に極めて有利である。
After 72 hours, pyruvic acid in the culture solution was quantified.2
It was 0.5g/Q, and there was no residual sugar. <Effects of the Invention> The method of the present invention makes it possible to accumulate a significant amount of pyruvic acid by fermentation, obtain pyruvic acid in high yield and yield, and generate almost no by-products. It is extremely advantageous industrially.

Claims (3)

【特許請求の範囲】[Claims] (1)トルロプシス(Torulopsis)属に属し
、生育のためにチアミンを必要とするピルビン酸生産菌
を培養することにより培地中にピルビン酸を生成蓄積さ
せ、これを採取することを特徴とする発酵法によるピル
ビン酸の製造法。
(1) A fermentation method characterized by producing and accumulating pyruvate in a medium by culturing a pyruvate-producing bacterium that belongs to the genus Torulopsis and requires thiamine for growth, and collecting it. Method for producing pyruvic acid by.
(2)ピルビン酸生産菌が、生育のためにさらにビオチ
ンを必要とする特許請求の範囲第1項記載の発酵法によ
るピルビン酸の製造法。
(2) A method for producing pyruvic acid by the fermentation method according to claim 1, wherein the pyruvic acid-producing bacteria further requires biotin for growth.
(3)ビルピン酸生産菌がトルロプシス(Torulo
psis)属グラブラータ(glabrata)種また
はトルロプシス(Torulopsis)属メタノロベ
ッセンス(methanolovescens)種に属
する特許請求の範囲第1項記載の発酵法によるピルビン
酸の製造法。
(3) Virupic acid producing bacteria are Torulopsis (Torulo).
A method for producing pyruvic acid by the fermentation method according to claim 1, which belongs to the genus Torulopsis glabrata species or the genus Torulopsis species methanolovescens.
JP27531086A 1986-02-27 1986-11-20 Production of pyruvic acid by fermentation Granted JPS62275688A (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
JP4258386 1986-02-27
JP61-42583 1986-02-27

Publications (2)

Publication Number Publication Date
JPS62275688A true JPS62275688A (en) 1987-11-30
JPH0358275B2 JPH0358275B2 (en) 1991-09-04

Family

ID=12640089

Family Applications (1)

Application Number Title Priority Date Filing Date
JP27531086A Granted JPS62275688A (en) 1986-02-27 1986-11-20 Production of pyruvic acid by fermentation

Country Status (1)

Country Link
JP (1) JPS62275688A (en)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0389620A1 (en) * 1987-08-21 1990-10-03 Toray Industries, Inc. Process for preparing pyruvic acid by fermentation

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS6214789A (en) * 1985-07-12 1987-01-23 Natl Food Res Inst Production of pyruvic acid
JPS635080A (en) * 1986-06-25 1988-01-11 バイエル・アクチエンゲゼルシヤフト 2-trifluoromethyl-benzimidazole

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS6214789A (en) * 1985-07-12 1987-01-23 Natl Food Res Inst Production of pyruvic acid
JPS635080A (en) * 1986-06-25 1988-01-11 バイエル・アクチエンゲゼルシヤフト 2-trifluoromethyl-benzimidazole

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0389620A1 (en) * 1987-08-21 1990-10-03 Toray Industries, Inc. Process for preparing pyruvic acid by fermentation

Also Published As

Publication number Publication date
JPH0358275B2 (en) 1991-09-04

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