JPH0424987B2 - - Google Patents
Info
- Publication number
- JPH0424987B2 JPH0424987B2 JP21424784A JP21424784A JPH0424987B2 JP H0424987 B2 JPH0424987 B2 JP H0424987B2 JP 21424784 A JP21424784 A JP 21424784A JP 21424784 A JP21424784 A JP 21424784A JP H0424987 B2 JPH0424987 B2 JP H0424987B2
- Authority
- JP
- Japan
- Prior art keywords
- isobutyric acid
- methanol
- culture
- strain
- growth
- 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
Links
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 27
- KQNPFQTWMSNSAP-UHFFFAOYSA-N isobutyric acid Chemical compound CC(C)C(O)=O KQNPFQTWMSNSAP-UHFFFAOYSA-N 0.000 claims description 26
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 7
- 229910052799 carbon Inorganic materials 0.000 claims description 7
- 241000222120 Candida <Saccharomycetales> Species 0.000 claims description 6
- 238000004519 manufacturing process Methods 0.000 claims description 6
- 244000005700 microbiome Species 0.000 claims description 4
- 238000012258 culturing Methods 0.000 claims description 2
- 239000002609 medium Substances 0.000 description 7
- 241000222124 [Candida] boidinii Species 0.000 description 5
- 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 description 4
- 150000001720 carbohydrates Chemical class 0.000 description 4
- 241000894006 Bacteria Species 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 150000003863 ammonium salts Chemical class 0.000 description 2
- 230000001580 bacterial effect Effects 0.000 description 2
- 229960002685 biotin Drugs 0.000 description 2
- 235000020958 biotin Nutrition 0.000 description 2
- 239000011616 biotin Substances 0.000 description 2
- 230000034303 cell budding Effects 0.000 description 2
- XBDQKXXYIPTUBI-UHFFFAOYSA-N dimethylselenoniopropionate Natural products CCC(O)=O XBDQKXXYIPTUBI-UHFFFAOYSA-N 0.000 description 2
- OVBPIULPVIDEAO-LBPRGKRZSA-N folic acid Chemical compound C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 OVBPIULPVIDEAO-LBPRGKRZSA-N 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 230000001766 physiological effect Effects 0.000 description 2
- 239000002244 precipitate Substances 0.000 description 2
- 239000002689 soil Substances 0.000 description 2
- 241000894007 species Species 0.000 description 2
- WLAMNBDJUVNPJU-UHFFFAOYSA-N 2-methylbutyric acid Chemical compound CCC(C)C(O)=O WLAMNBDJUVNPJU-UHFFFAOYSA-N 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- 241000512933 Candida cariosilignicola Species 0.000 description 1
- 241000177202 Chimonobambusa utilis Species 0.000 description 1
- 108010010803 Gelatin Proteins 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
- OVBPIULPVIDEAO-UHFFFAOYSA-N N-Pteroyl-L-glutaminsaeure Natural products C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)NC(CCC(O)=O)C(O)=O)C=C1 OVBPIULPVIDEAO-UHFFFAOYSA-N 0.000 description 1
- 229910002651 NO3 Inorganic materials 0.000 description 1
- NHNBFGGVMKEFGY-UHFFFAOYSA-N Nitrate Chemical compound [O-][N+]([O-])=O NHNBFGGVMKEFGY-UHFFFAOYSA-N 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 1
- 241000235015 Yarrowia lipolytica Species 0.000 description 1
- 241000512904 [Candida] succiphila Species 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 239000004202 carbamide Substances 0.000 description 1
- 235000014633 carbohydrates Nutrition 0.000 description 1
- 235000021466 carotenoid Nutrition 0.000 description 1
- 150000001747 carotenoids Chemical class 0.000 description 1
- 230000000052 comparative effect Effects 0.000 description 1
- ARUVKPQLZAKDPS-UHFFFAOYSA-L copper(II) sulfate Chemical compound [Cu+2].[O-][S+2]([O-])([O-])[O-] ARUVKPQLZAKDPS-UHFFFAOYSA-L 0.000 description 1
- 229910000366 copper(II) sulfate Inorganic materials 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- 229960000304 folic acid Drugs 0.000 description 1
- 235000019152 folic acid Nutrition 0.000 description 1
- 239000011724 folic acid Substances 0.000 description 1
- 238000002290 gas chromatography-mass spectrometry Methods 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 239000002932 luster Substances 0.000 description 1
- 238000000034 method Methods 0.000 description 1
- 230000000877 morphologic effect Effects 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- 235000005985 organic acids Nutrition 0.000 description 1
- 150000002894 organic compounds Chemical class 0.000 description 1
- 239000001814 pectin Substances 0.000 description 1
- 235000010987 pectin Nutrition 0.000 description 1
- 229920001277 pectin Polymers 0.000 description 1
- 235000021317 phosphate Nutrition 0.000 description 1
- 150000003013 phosphoric acid derivatives Chemical class 0.000 description 1
- 239000000049 pigment Substances 0.000 description 1
- 230000002062 proliferating effect Effects 0.000 description 1
- 235000019260 propionic acid Nutrition 0.000 description 1
- ZUFQODAHGAHPFQ-UHFFFAOYSA-N pyridoxine hydrochloride Chemical compound Cl.CC1=NC=C(CO)C(CO)=C1O ZUFQODAHGAHPFQ-UHFFFAOYSA-N 0.000 description 1
- 229960004172 pyridoxine hydrochloride Drugs 0.000 description 1
- 235000019171 pyridoxine hydrochloride Nutrition 0.000 description 1
- 239000011764 pyridoxine hydrochloride Substances 0.000 description 1
- IUVKMZGDUIUOCP-BTNSXGMBSA-N quinbolone Chemical compound O([C@H]1CC[C@H]2[C@H]3[C@@H]([C@]4(C=CC(=O)C=C4CC3)C)CC[C@@]21C)C1=CCCC1 IUVKMZGDUIUOCP-BTNSXGMBSA-N 0.000 description 1
- 229940052714 riboflavin 1 mg Drugs 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- GQTHJBOWLPZUOI-FJXQXJEOSA-M sodium D-pantothenate Chemical compound [Na+].OCC(C)(C)[C@@H](O)C(=O)NCCC([O-])=O GQTHJBOWLPZUOI-FJXQXJEOSA-M 0.000 description 1
- 235000019188 sodium D-pantothenate Nutrition 0.000 description 1
- 239000011756 sodium D-pantothenate Substances 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 150000003467 sulfuric acid derivatives Chemical class 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
- 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
- UAXOELSVPTZZQG-UHFFFAOYSA-N tiglic acid Natural products CC(C)=C(C)C(O)=O UAXOELSVPTZZQG-UHFFFAOYSA-N 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 150000003722 vitamin derivatives Chemical class 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
- NWONKYPBYAMBJT-UHFFFAOYSA-L zinc sulfate Chemical compound [Zn+2].[O-]S([O-])(=O)=O NWONKYPBYAMBJT-UHFFFAOYSA-L 0.000 description 1
- 229910000368 zinc sulfate Inorganic materials 0.000 description 1
- 239000011686 zinc sulphate Substances 0.000 description 1
- 235000009529 zinc sulphate Nutrition 0.000 description 1
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Description
（産業上の利用分野）
本発明はイソ酪酸の製造方法に関する。
（発明の目的）
本発明者らは、メタノールを炭素源とするイソ
酪酸の製造方法について種々検討した結果、カン
デイダ属に属する微生物がメタノールをイソ酪酸
に変換する能力を有することを見出し、本発明に
到達した。
すなわち、本発明の要旨は、カンデイダ属に属
し、イソ酪酸を生産する能力を有する微生物を、
炭素源としてメタノールを使用して培養し、培養
物からイソ酪酸を得ることを特徴とするイソ酪酸
の製造方法にある。
（発明の構成）
以下、本発明を詳細に説明する。
本発明において使用される微生物はカンデイダ
（Candida）属に属し、イソ酪酸を生産する能力
を有するものであり、たとえば、カンデイダ ボ
イデイニイ（Candida boidinii）Ａ−1917−３
（微工研菌寄第7853号）が挙げられる。
この菌株は、土壌より分離されたものであり、
その菌学的性状は次の通りである。
各培地における生育状態
(1) YM液体培地、30℃、３日間の生育状態
生育は良好で培養液面に環状の皮膜が形成
される。培養液底部に沈澱を生ずる。
(2) YM平板寒天上の性質
30℃、３日間の培養で、生育は良好。コロ
ニーは、バター質、白色を呈する。
コロニー表面は平滑；周縁は全縁〜やや波
状、隆起状態は台状；輝光状の光沢を呈す
る。細胞形態は、卵形〜楕円形、大きさ２−
10.3×1.3−3.3（平均3.6×2.5）μｍ、増殖様
式は多極出芽、偽菌糸を形成する。
子のう胞子は形成されない。
(3) 1.5％メタノール含有液体培地、30℃、３
日間の生育状態。
生育は良好で培養液面に皮膜は殆ど形成さ
れない。培養液底部に少量の沈澱が形成され
る。
生理的性質
生育の範囲 PH２〜９
温度 10〜30℃37℃で生育せず。
硝酸塩の同化：あり
ゼラチン液化能：なし
デンプン類似物質の生成：なし
50％グルコース上での生育（YM）：なし
NaCl耐性：８−９％
メタノール資化性：あり
ペクチン資化性：あり
ビタミン要求性：ビオチンパントテン酸−
Ca
(Industrial Application Field) The present invention relates to a method for producing isobutyric acid. (Object of the Invention) As a result of various studies on the production method of isobutyric acid using methanol as a carbon source, the present inventors discovered that microorganisms belonging to the genus Candeida have the ability to convert methanol into isobutyric acid. reached. That is, the gist of the present invention is to use a microorganism that belongs to the genus Candida and has the ability to produce isobutyric acid.
A method for producing isobutyric acid, which comprises culturing using methanol as a carbon source and obtaining isobutyric acid from the culture. (Structure of the Invention) The present invention will be described in detail below. The microorganism used in the present invention belongs to the genus Candida and has the ability to produce isobutyric acid, for example, Candida boidinii A-1917-3.
(Feikoken Bibori No. 7853). This strain was isolated from soil,
Its mycological properties are as follows. Growth status in each medium (1) Growth status in YM liquid medium, 30℃, 3 days Growth is good, with a ring-shaped film forming on the surface of the culture medium. A precipitate forms at the bottom of the culture solution. (2) Properties on YM plate agar Growth is good when cultured at 30℃ for 3 days. Colonies are buttery and white in color. The colony surface is smooth; the periphery is entirely to slightly wavy, and the ridges are plate-shaped; it exhibits a luminous luster. Cell morphology is oval to oval, size 2-
10.3 x 1.3-3.3 (average 3.6 x 2.5) μm, growth mode is multipolar budding, forming pseudohyphae. Ascospores are not formed. (3) Liquid medium containing 1.5% methanol, 30℃, 3
Daily growth status. Growth is good and almost no film is formed on the surface of the culture solution. A small amount of precipitate will form at the bottom of the culture solution. Physiological properties Growth range PH2-9 Temperature 10-30℃, does not grow at 37℃. Assimilation of nitrate: Yes Gelatin liquefaction ability: No Production of starch-like substances: No Growth on 50% glucose (YM): No NaCl tolerance: 8-9% Methanol assimilation: Yes Pectin assimilation: Yes Vitamin requirement Characteristic: biotin pantothenic acid-
Ca
【表】【table】
【表】【table】
【表】
属レベルの同定
本菌株（Ａ−1917−３）は、カロチノイド色
素を生産しない、偽菌糸を常に形成する、多極
出芽によつて増殖する、子のう胞子を形成しな
いことからロダーの“ザ・イースト”（２版・
1970）に記載されているカンデイダ
（Candida）属に帰属することが判明した。
種の同定
リー及び駒形（Lee＆Komagata）（1980）
のメタノール資化菌の分類学的研究
“Taxonomical study of methanol−
assimilatingyedsts”によればカンデイダ属の
中にメタノール資化菌種として５種を認めてい
る。（C.boidinii，C.cariosilignicola，C.
lipolytica，C.succiphila，C.utilis）。
これらの５種は糖類の発酵能パターン及び糖
類の資化性パターンによつて主に識別されてい
る。
本株菌（Ａ−1917−３）の糖類の発酵性試験
及び上記の炭素源の資化性パターンはロダー
（1970）のザ・イースト及びリーと駒形（1980）
によつて記載されているカンデイダ・ボイデイ
ニイ（Candida boidinii）の諸性質とよく合致
する。
よつて本菌株（Ａ−1917−３）はカンデイ
ダ・ボイデイニイ（Candida boidinii）と同定
された。また本菌株（Ａ−1917−３）とカンデ
イダ・ボイデイニイ（Candida boidinii）の
Authentic strain（IAM12269株）の形態上の特
徴、糖類の発酵能、炭素源の資化性及び他の生
理的性質について比較検討したところ、本菌株
（Ａ−1917−３）の諸性質はカンデイダ・ボイ
デイニイ（IAM12269）のそれらともよく一致
した。
本発明において使用される培地としては、主炭
素源としてメタノールを含むものであれば、特に
制限されない。
炭素源としては、メタノール以外に、種々の炭
水化物、有機酸等をさらに添加してもよく、窒素
源としては、有機アンモニウム塩、無機アンモニ
ウム塩、尿素等を用いることができる。
また、必要に応じ、無機物として各種リン酸
塩、硫酸塩等を使用することができ、必要に応じ
各種有機栄養物を添加することもできる。
培養は、通常12時間〜14日間程度、好気的条件
下に行なわれる。
培地のPHは４−10、温度は20−40℃程度から選
ばれる。
イソ酪酸の生産に際しては、増殖菌体、休止菌
体のいずれをも用いることができる。
培養物からイソ酪酸の採取、精製に際しては一
般に有機化合物の採取、精製に用いられている方
法を採用することができる。
（実施例）
以下、実施例により、本発明をさらに説明す
る。
なお、実施例における物質の同定はガスクロマ
トグラフ−質量分析等により標品と比較して行な
つた。
実施例 １
メタノール40ｇ、NH4Cl4ｇ、K2HPO41ｇ、
Na2HPO41ｇ、MgSO4・7H2O0.5ｇ、塩酸ピリ
ドキシン１mg、ビオチン0.01mg、塩酸チアミン１
mg、リボフラビン１mg、Ｄ−パントテン酸ナトリ
ウム１mg、葉酸0.01mg、FeSO4・7H2O1mg、
ZnSO4・7H2O1mg、CuSO4・5H2O0.1mg、
MnCl2・4H2O0.04mg、水１からなるPH7.0の培
地を調整した。この培地50mlを500ml容コルベン
に分注し、120℃で10分間殺菌した。
天然の土壌から分離したカンデイダ・ボイデイ
ニイＡ−1917−３菌の斜面培養（30℃、３〜６日
間）菌の一白金耳を上記コルベンに接種し、112
往復／分の往復振とう機で30℃10日間培養を行な
い、この培養物50mlよりイソ酪酸1.5mgを得た
（同時にプロピオン酸、イソ吉草酪、２−メチル
酪酸を得た）。[Table] Identification at the genus level This strain (A-1917-3) does not produce carotenoid pigments, always forms pseudohyphae, grows by multipolar budding, and does not form ascospores. “The East” (2nd edition/
It was found that it belonged to the genus Candida, which was described in 1970). Species identification Lee & Komagata (1980)
“Taxonomical study of methanol−
According to "Assimilating Yedsts", five species of methanol-assimilating bacteria are recognized within the genus Candeida (C.boidinii, C.cariosilignicola, C.
lipolytica, C. succiphila, C. utilis). These five types are mainly distinguished by their saccharide fermentability patterns and saccharide assimilation patterns. The saccharide fermentability test of this strain (A-1917-3) and the carbon source assimilation pattern described above are based on the yeast of Roder (1970) and Lee and Komagata (1980).
This agrees well with the properties of Candida boidinii described by. Therefore, this strain (A-1917-3) was identified as Candida boidinii. In addition, this strain (A-1917-3) and Candida boidinii
A comparative study of the morphological characteristics, sugar fermentation ability, carbon source assimilation ability, and other physiological properties of the authentic strain (IAM12269 strain) revealed that the properties of this strain (A-1917-3) were similar to that of Candida. It also matched well with those of Boideinii (IAM12269). The medium used in the present invention is not particularly limited as long as it contains methanol as a main carbon source. As a carbon source, in addition to methanol, various carbohydrates, organic acids, etc. may be further added, and as a nitrogen source, organic ammonium salts, inorganic ammonium salts, urea, etc. can be used. Furthermore, various phosphates, sulfates, etc. can be used as inorganic substances, and various organic nutrients can also be added as necessary. Cultivation is usually carried out under aerobic conditions for about 12 hours to 14 days. The pH of the medium is selected from 4-10, and the temperature is selected from about 20-40°C. In the production of isobutyric acid, both proliferating bacterial cells and dormant bacterial cells can be used. When collecting and purifying isobutyric acid from the culture, methods generally used for collecting and purifying organic compounds can be employed. (Example) Hereinafter, the present invention will be further explained with reference to Examples. The substances in the Examples were identified by comparing them with standard samples by gas chromatography-mass spectrometry and the like. Example 1 Methanol 40g, NH 4 Cl4g, K 2 HPO 4 1g,
Na 2 HPO 4 1g, MgSO 4・7H 2 O 0.5g, pyridoxine hydrochloride 1mg, biotin 0.01mg, thiamine hydrochloride 1
mg, riboflavin 1 mg, sodium D-pantothenate 1 mg, folic acid 0.01 mg, FeSO 4 7H 2 O 1 mg,
ZnSO4・7H2O1mg , CuSO4・5H2O0.1mg ,
A medium with a pH of 7.0 consisting of 0.04 mg of MnCl 2 4H 2 O and 1 part water was prepared. 50 ml of this medium was dispensed into a 500 ml Colben and sterilized at 120°C for 10 minutes. Slant culture of Candida boideinii A-1917-3 bacteria isolated from natural soil (30°C, 3-6 days) A loopful of the bacteria was inoculated into the above Colben.
Culture was carried out at 30°C for 10 days using a reciprocating shaker at a reciprocating rate of 10 days, and 1.5 mg of isobutyric acid was obtained from 50 ml of this culture (propionic acid, isovalerbutyric acid, and 2-methylbutyric acid were also obtained).
Claims (1)
を生産する能力を有する微生物を、炭素源として
メタノールを使用して培養し、培養物からイソ酪
酸を得ることを特徴とするイソ酪酸の製造方法。1. A method for producing isobutyric acid, which comprises culturing a microorganism belonging to the genus Candida and having the ability to produce isobutyric acid using methanol as a carbon source, and obtaining isobutyric acid from the culture.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP21424784A JPS6192581A (en) | 1984-10-15 | 1984-10-15 | Production of isobutyric acid |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP21424784A JPS6192581A (en) | 1984-10-15 | 1984-10-15 | Production of isobutyric acid |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPS6192581A JPS6192581A (en) | 1986-05-10 |
| JPH0424987B2 true JPH0424987B2 (en) | 1992-04-28 |
Family
ID=16652605
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP21424784A Granted JPS6192581A (en) | 1984-10-15 | 1984-10-15 | Production of isobutyric acid |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS6192581A (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0197589A (en) * | 1987-10-07 | 1989-04-17 | Fujitsu Ltd | Determination system of fundamental attitude of multi-joint robot |
| CN117821274A (en) * | 2023-12-22 | 2024-04-05 | 华南师范大学 | Application of candida boidinii in denitrification |
-
1984
- 1984-10-15 JP JP21424784A patent/JPS6192581A/en active Granted
Also Published As
| Publication number | Publication date |
|---|---|
| JPS6192581A (en) | 1986-05-10 |
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