JPH0358275B2 - - Google Patents
Info
- Publication number
- JPH0358275B2 JPH0358275B2 JP61275310A JP27531086A JPH0358275B2 JP H0358275 B2 JPH0358275 B2 JP H0358275B2 JP 61275310 A JP61275310 A JP 61275310A JP 27531086 A JP27531086 A JP 27531086A JP H0358275 B2 JPH0358275 B2 JP H0358275B2
- Authority
- JP
- Japan
- Prior art keywords
- pyruvate
- producing
- pyruvic acid
- biotin
- torulopsis
- 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.)
- Expired - Lifetime
Links
- LCTONWCANYUPML-UHFFFAOYSA-N Pyruvic acid Chemical compound CC(=O)C(O)=O LCTONWCANYUPML-UHFFFAOYSA-N 0.000 claims description 20
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 claims description 18
- LCTONWCANYUPML-UHFFFAOYSA-M Pyruvate Chemical compound CC(=O)C([O-])=O LCTONWCANYUPML-UHFFFAOYSA-M 0.000 claims description 14
- 229940076788 pyruvate Drugs 0.000 claims description 14
- 238000000034 method Methods 0.000 claims description 10
- 229940107700 pyruvic acid Drugs 0.000 claims description 10
- JZRWCGZRTZMZEH-UHFFFAOYSA-N Thiamine Natural products CC1=C(CCO)SC=[N+]1CC1=CN=C(C)N=C1N JZRWCGZRTZMZEH-UHFFFAOYSA-N 0.000 claims description 9
- 229960002685 biotin Drugs 0.000 claims description 9
- 235000020958 biotin Nutrition 0.000 claims description 9
- 239000011616 biotin Substances 0.000 claims description 9
- 241000222120 Candida <Saccharomycetales> Species 0.000 claims description 8
- 238000000855 fermentation Methods 0.000 claims description 8
- 230000004151 fermentation Effects 0.000 claims description 8
- 241000894006 Bacteria Species 0.000 claims description 7
- 235000019157 thiamine Nutrition 0.000 claims description 7
- 239000011721 thiamine Substances 0.000 claims description 7
- 238000012258 culturing Methods 0.000 claims description 5
- 238000004519 manufacturing process Methods 0.000 claims description 5
- KYMBYSLLVAOCFI-UHFFFAOYSA-N thiamine Chemical compound CC1=C(CCO)SCN1CC1=CN=C(C)N=C1N KYMBYSLLVAOCFI-UHFFFAOYSA-N 0.000 claims description 5
- 229960003495 thiamine Drugs 0.000 claims description 5
- 241000222126 [Candida] glabrata Species 0.000 claims description 4
- PVNIIMVLHYAWGP-UHFFFAOYSA-N Niacin Chemical compound OC(=O)C1=CC=CN=C1 PVNIIMVLHYAWGP-UHFFFAOYSA-N 0.000 description 7
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 6
- LXNHXLLTXMVWPM-UHFFFAOYSA-N pyridoxine Chemical compound CC1=NC=C(CO)C(CO)=C1O LXNHXLLTXMVWPM-UHFFFAOYSA-N 0.000 description 6
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 5
- 229960003512 nicotinic acid Drugs 0.000 description 5
- 235000001968 nicotinic acid Nutrition 0.000 description 5
- 239000011664 nicotinic acid Substances 0.000 description 5
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 4
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- 239000001888 Peptone Substances 0.000 description 3
- 108010080698 Peptones Proteins 0.000 description 3
- KWYUFKZDYYNOTN-UHFFFAOYSA-M Potassium hydroxide Chemical compound [OH-].[K+] KWYUFKZDYYNOTN-UHFFFAOYSA-M 0.000 description 3
- 239000006227 byproduct Substances 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 235000019319 peptone Nutrition 0.000 description 3
- 235000008160 pyridoxine Nutrition 0.000 description 3
- 239000011677 pyridoxine Substances 0.000 description 3
- 239000002994 raw material Substances 0.000 description 3
- 150000003839 salts Chemical class 0.000 description 3
- 229940088594 vitamin Drugs 0.000 description 3
- 229930003231 vitamin Natural products 0.000 description 3
- 235000013343 vitamin Nutrition 0.000 description 3
- 239000011782 vitamin Substances 0.000 description 3
- 229940011671 vitamin b6 Drugs 0.000 description 3
- NLXLAEXVIDQMFP-UHFFFAOYSA-N Ammonia chloride Chemical compound [NH4+].[Cl-] NLXLAEXVIDQMFP-UHFFFAOYSA-N 0.000 description 2
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 2
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-N 0.000 description 2
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 2
- 229940024606 amino acid Drugs 0.000 description 2
- 150000001413 amino acids Chemical class 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 229910052799 carbon Inorganic materials 0.000 description 2
- 238000007796 conventional method Methods 0.000 description 2
- 239000000284 extract Substances 0.000 description 2
- 229910052742 iron Inorganic materials 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Chemical compound [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 description 2
- ZUFQODAHGAHPFQ-UHFFFAOYSA-N pyridoxine hydrochloride Chemical compound Cl.CC1=NC=C(CO)C(CO)=C1O ZUFQODAHGAHPFQ-UHFFFAOYSA-N 0.000 description 2
- 229960004172 pyridoxine hydrochloride Drugs 0.000 description 2
- 235000019171 pyridoxine hydrochloride Nutrition 0.000 description 2
- 239000011764 pyridoxine hydrochloride Substances 0.000 description 2
- 235000011121 sodium hydroxide Nutrition 0.000 description 2
- DAEPDZWVDSPTHF-UHFFFAOYSA-M sodium pyruvate Chemical compound [Na+].CC(=O)C([O-])=O DAEPDZWVDSPTHF-UHFFFAOYSA-M 0.000 description 2
- 230000001954 sterilising effect Effects 0.000 description 2
- 238000004659 sterilization and disinfection Methods 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- LWIHDJKSTIGBAC-UHFFFAOYSA-K tripotassium phosphate Chemical compound [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 description 2
- PAWQVTBBRAZDMG-UHFFFAOYSA-N 2-(3-bromo-2-fluorophenyl)acetic acid Chemical compound OC(=O)CC1=CC=CC(Br)=C1F PAWQVTBBRAZDMG-UHFFFAOYSA-N 0.000 description 1
- PWKSKIMOESPYIA-UHFFFAOYSA-N 2-acetamido-3-sulfanylpropanoic acid Chemical compound CC(=O)NC(CS)C(O)=O PWKSKIMOESPYIA-UHFFFAOYSA-N 0.000 description 1
- 241000221198 Basidiomycota Species 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 1
- 240000008042 Zea mays Species 0.000 description 1
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 1
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 239000003905 agrochemical Substances 0.000 description 1
- 239000003513 alkali Substances 0.000 description 1
- 235000019270 ammonium chloride Nutrition 0.000 description 1
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 1
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 1
- 235000011130 ammonium sulphate Nutrition 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- 229910000019 calcium carbonate Inorganic materials 0.000 description 1
- 235000010216 calcium carbonate Nutrition 0.000 description 1
- 239000004202 carbamide Substances 0.000 description 1
- 235000013877 carbamide Nutrition 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 235000014633 carbohydrates Nutrition 0.000 description 1
- 238000012790 confirmation Methods 0.000 description 1
- 235000005822 corn Nutrition 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000006911 enzymatic reaction Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 239000000796 flavoring agent Substances 0.000 description 1
- 235000019634 flavors Nutrition 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 108010067653 lactate dehydratase Proteins 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 238000004811 liquid chromatography Methods 0.000 description 1
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 1
- 235000019341 magnesium sulphate Nutrition 0.000 description 1
- WPBNNNQJVZRUHP-UHFFFAOYSA-L manganese(2+);methyl n-[[2-(methoxycarbonylcarbamothioylamino)phenyl]carbamothioyl]carbamate;n-[2-(sulfidocarbothioylamino)ethyl]carbamodithioate Chemical compound [Mn+2].[S-]C(=S)NCCNC([S-])=S.COC(=O)NC(=S)NC1=CC=CC=C1NC(=S)NC(=O)OC WPBNNNQJVZRUHP-UHFFFAOYSA-L 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 229930014626 natural product Natural products 0.000 description 1
- 229910017464 nitrogen compound Inorganic materials 0.000 description 1
- 150000002830 nitrogen compounds Chemical class 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- 235000005985 organic acids Nutrition 0.000 description 1
- 150000002897 organic nitrogen compounds Chemical class 0.000 description 1
- 229910052697 platinum Inorganic materials 0.000 description 1
- 235000011118 potassium hydroxide Nutrition 0.000 description 1
- 229910000160 potassium phosphate Inorganic materials 0.000 description 1
- 235000011009 potassium phosphates Nutrition 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 238000011002 quantification Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- IFGCUJZIWBUILZ-UHFFFAOYSA-N sodium 2-[[2-[[hydroxy-(3,4,5-trihydroxy-6-methyloxan-2-yl)oxyphosphoryl]amino]-4-methylpentanoyl]amino]-3-(1H-indol-3-yl)propanoic acid Chemical compound [Na+].C=1NC2=CC=CC=C2C=1CC(C(O)=O)NC(=O)C(CC(C)C)NP(O)(=O)OC1OC(C)C(O)C(O)C1O IFGCUJZIWBUILZ-UHFFFAOYSA-N 0.000 description 1
- 229940054269 sodium pyruvate Drugs 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- DPJRMOMPQZCRJU-UHFFFAOYSA-M thiamine hydrochloride Chemical compound Cl.[Cl-].CC1=C(CCO)SC=[N+]1CC1=CN=C(C)N=C1N DPJRMOMPQZCRJU-UHFFFAOYSA-M 0.000 description 1
- 229960000344 thiamine hydrochloride Drugs 0.000 description 1
- 235000019190 thiamine hydrochloride Nutrition 0.000 description 1
- 239000011747 thiamine hydrochloride Substances 0.000 description 1
- 229960004441 tyrosine Drugs 0.000 description 1
- 239000012138 yeast extract Substances 0.000 description 1
Description
【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
するものである。DETAILED DESCRIPTION OF THE INVENTION <Industrial Field of 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、
各種医・農薬などの有効な合成原料であるのみな
らず酵素法によるＬ−トリプトフアン、Ｌ−シス
テイン、Ｌ−チロシンなどのアミノ酸合成の主要
原料である。よつて安価に製造し得れば、種々の
合成原料として有用である。 Pyruvate is an important intermediate in biological metabolism,
It is not only an effective raw material for the synthesis of various medicines and agricultural chemicals, but also a main raw material for the synthesis of amino acids such as L-tryptophan, L-cysteine, and L-tyrosine by enzymatic methods. Therefore, if it can be produced at low cost, it is 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ヤンデイダ属など
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57−159492号公報等）。<Prior art> Conventionally, as a method for producing pyruvic acid by a fermentation method, a method using yeasts such as Satucharomyces and Candeida, basidiomycetes, or special bacteria is known (Japanese Patent Publication No. 57-796,
(Japanese Unexamined Patent Publication No. 159492/1983, etc.).
＜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
つたり、または、収率・収量が十分でなかつたり
したため工業的に有利な方法とは言えなかつた。<Problems to be Solved by the Present Invention> However, such conventional methods cannot be said to be industrially advantageous because they produce a large amount of by-products or the yield/yield is insufficient.
＜問題点を解決するための手段および作用＞
したがつ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、トルロプシス属に属し、生育のためにチア
ミンおよびビオチンを必要と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 effects for solving the problems> Therefore, the present inventors conducted intensive research with the aim of finding a novel pyruvate-producing bacterium using a fermentation method that can solve the above problems. The present inventors have discovered that yeast, which belongs to the genus Torulopsis and requires thiamine and biotin for growth, exhibits excellent effects, and has thus arrived at the present invention.
すなわち、本発明の上記目的は、トルロプシス
属に属し、生育のためにチアミンおよびビオチン
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
採取することにより、達成されるのである。 That is, the above object of the present invention is to produce and accumulate pyruvate in a medium by culturing pyruvate-producing bacteria that belong to the genus Torulopsis and require thiamine and biotin for growth, and to collect the pyruvate. , will be achieved.
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
つた。 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.
本発明に用いられるピルビン酸生産菌はピルビ
ン酸生成能を有するトルロプシス属に属する酵母
菌であつて生育のためにチアミンおよびビオチン
を必要とするものであれば、いかなるものであつ
てもよい。さらに、他の各種のビタミン、例えば
ナイアシン、ピリドキシンなどを単独もしくは併
せて要求するものであつてもよい。 The pyruvate-producing bacteria used in the present invention may be any yeast that belongs to the genus Torulopsis and has the ability to produce pyruvate and requires thiamine and biotin for growth. Furthermore, various other vitamins such as niacin and pyridoxine may be required alone or in combination.
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）（IFO 0005）（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・メタノロベツセンス
（Torulopsis methanolovescens）（ATCC26176）
（チアミンおよびビオチン要求）などが挙げられ
る。 Examples of pyruvate-producing bacteria used in the present invention include Torulopsis glabrata (IFO 0005) (thiamine,
biotin, pyridoxine and niacin requirements),
Torulopsis methanolovescens (ATCC26176)
(thiamin and biotin requirements), etc.
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、硫
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、さ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。 The medium used in the present invention may contain moderate amounts of carbon sources, nitrogen sources, inorganic salts, vitamins, etc. commonly used in fermentation, but carbon sources include carbohydrates such as glucose, organic acids, Ethanol, methanol, and other substances that 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 sulfate, 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 extracts and corn extracts containing these are used. A medium supplemented with 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、PH
の低下が起こるので炭酸カルシウム、苛性ソー
ダ、苛性カリなどのアルカリでPH３〜７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℃〜32℃が適当である。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、フエ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 culture, as pyruvate is produced and accumulated, the pH decreases.
Therefore, adjusting the pH to 3 to 7 with an alkali such as calcium carbonate, caustic soda, or caustic potash is effective for pyruvic acid production. A suitable temperature during culturing is 22°C to 32°C. After culturing,
Pyruvate accumulated in the system can be isolated and collected by conventional methods. For example, methods such as acidic ether extraction, phenylhydrazonation, and precipitation isolation may also be employed.
＜実施例＞ 以下、実施例によつ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イーによる方法およ
び乳酸脱水酵素による方法によつて行つた。以下
の分析結果については上記両分析法ともよく合致
しており、同じ分析数値を示した。 Confirmation and quantification of pyruvic acid produced in the Examples were performed by a method using high performance liquid chromatography and a method using lactate dehydratase. 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
グリコース10％、ペプトン１％、KH2P040.1
％、MzSO4・7H2O0.05％、Fe2＋2PPm、チアミ
ン塩酸塩2μｇ／、ピリドキシン塩酸塩100μ
ｇ／、ニコチン酸100μｇ／、PH5.0からなる
培地30mlを500ml容振盪フラスコに分注滅菌後、
別に滅菌したCaCO34％を添加し、トルロプシ
ス・グラブラータ（IFC 0005）を１白金耳植菌
した後、30℃で91時間振盪培養した。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あ
つた。培養後ピルビン酸を定量したところ40.7
ｇ／であり、残糖は22ｇ／であつた。副生物
は液体クロマトグラフイーでは検出されなかつ
た。この培養液１を除菌後、上澄液に塩酸を加
えPH2.0とし、１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100ml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23.7ｇ（純度
97％）を得た。Example 1 Glycose 10%, peptone 1%, KH 2 P0 4 0.1
%, MzSO 4・7H 2 O 0.05%, Fe 2 +2PPm, thiamine hydrochloride 2μg/, pyridoxine hydrochloride 100μ
After sterilization, dispense 30 ml of a medium containing g/g/, 100 μg/nicotinic acid, and pH 5.0 into a 500 ml shaking flask.
Separately, 4% sterilized CaCO 3 was added, one platinum loop of Torulopsis glabrata (IFC 0005) was inoculated, and the mixture was cultured with shaking at 30° C. for 91 hours. Even without adding biotin, the amount contained in peptone was sufficient. After culturing, pyruvate was quantified and found to be 40.7.
g/, and the residual sugar was 22 g/. No by-products were detected by liquid chromatography. After sterilization of this culture solution 1, hydrochloric acid was added to the supernatant to adjust the pH to 2.0, and the mixture was extracted with ethyl ether from 1.
Next, the pH was neutralized to 6.0 with caustic soda and concentrated under reduced pressure at 40°C to a volume of about 100 ml. Ethanol was added dropwise to this concentrated solution and 23.7g of sodium pyruvate (purity
97%).
実施例 ２
実施例１において使用した培地中のグリコース
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0.5μｇ／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・メタノロベツセンス（ATCC
26176）に変えて実施例１と同様の方法で培養し
た。Example 2 Using the same medium as in Example 1 except that the glycose in the medium was reduced to 5% and the pyridoxine hydrochloride and nicotinic acid replaced with 0.5 μg/biotin, Torulopsis glabrata was grown into Torulopsis methanolobetsu. Sense (ATCC
26176) and cultured in the same manner as in Example 1.
72時間後、培養液中のピルビン酸を定量したと
ころ20.5ｇ／であり、残糖はなかつた。 After 72 hours, the amount of pyruvic acid in the culture solution was determined to be 20.5 g/day, and there was no residual sugar.
＜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。<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 (1)
育のためにチアミンおよびビオチンを必要とする
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。 ２ ピルビン酸生産菌がトルロプシス
（Torulopsis）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
ツセンス（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。[Scope of Claims] 1. Producing and accumulating pyruvate in a medium by culturing pyruvate-producing bacteria that belong to the genus Torulopsis and require thiamine and biotin for growth, and collecting the same. A method for producing pyruvic acid by a fermentation method, characterized by: 2. A method for producing pyruvic acid by the fermentation method according to claim 1, wherein the pyruvic acid-producing bacteria belong to the genus Torulopsis glabrata species or the genus Torulopsis species methanolovescens.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP61-42583 | 1986-02-27 | ||
| JP4258386 | 1986-02-27 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPS62275688A JPS62275688A (en) | 1987-11-30 |
| JPH0358275B2 true 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) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0389620B1 (en) * | 1987-08-21 | 1992-12-16 | Toray Industries, Inc. | Process for preparing pyruvic acid by fermentation |
Citations (2)
| 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 |
-
1986
- 1986-11-20 JP JP27531086A patent/JPS62275688A/en active Granted
Patent Citations (2)
| 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 |
Also Published As
| Publication number | Publication date |
|---|---|
| JPS62275688A (en) | 1987-11-30 |
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