JPH0424989B2 - - Google Patents
Info
- Publication number
- JPH0424989B2 JPH0424989B2 JP21424884A JP21424884A JPH0424989B2 JP H0424989 B2 JPH0424989 B2 JP H0424989B2 JP 21424884 A JP21424884 A JP 21424884A JP 21424884 A JP21424884 A JP 21424884A JP H0424989 B2 JPH0424989 B2 JP H0424989B2
- Authority
- JP
- Japan
- Prior art keywords
- methyl formate
- methanol
- growth
- strain
- culture
- 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
- TZIHFWKZFHZASV-UHFFFAOYSA-N methyl formate Chemical compound COC=O TZIHFWKZFHZASV-UHFFFAOYSA-N 0.000 claims description 26
- 241000222120 Candida <Saccharomycetales> Species 0.000 claims description 7
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 7
- 229910052799 carbon Inorganic materials 0.000 claims description 7
- 238000004519 manufacturing process Methods 0.000 claims description 6
- 244000005700 microbiome Species 0.000 claims description 4
- 238000012258 culturing Methods 0.000 claims 1
- 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
- 241000222124 [Candida] boidinii Species 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
- 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
- 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
- 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
- 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
- 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
- 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
- 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）
本発明はギ酸メチル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ギ酸
メチル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ギ酸メ
チル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、ギ酸メチル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、
培養物からギ酸メチルを得ることを特徴とするギ
酸メチルの製造方法にある。
（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、ギ酸メチルを生産する能
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 methyl formate. (Object of the Invention) As a result of various studies on the production method of methyl formate using methanol as a carbon source, the present inventors discovered that microorganisms belonging to the genus Candida have the ability to convert methanol into methyl formate. reached. That is, the gist of the present invention is to culture a microorganism belonging to the genus Candida and having the ability to produce methyl formate using methanol as a carbon source,
A method for producing methyl formate characterized by obtaining methyl formate from a 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 methyl formate.
Candida boidinii A-1917-3
(Feikoken Bibori No. 7853). This strain was isolated from soil, and its mycological properties are as follows. Growth status in each medium (1) Growth status in YM liquid medium, 30°C, 3 days. Growth is good, and a ring-shaped film is formed on the surface of the culture solution. A precipitate forms at the bottom of the culture solution. (2) Properties on YM plate agar When cultured at 30°C for 3 days, growth is good, and the colonies are buttery and white in color. The colony surface is smooth; the periphery is entirely to slightly wavy; the ridge is plate-like; it has a luminous luster. Cell morphology is oval to oval, size 2-10.3×1.3-
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 No growth at temperatures of 10-30℃ and 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。
本菌株（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 boidnii）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。
ギ酸メチルの生産に際しては、増殖菌体、休止
菌体のいずれをも用いることができる。
培養物からギ酸メチルの採取、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℃３日間培養を行ない、この培養
物50mlよりギ酸メチル２mg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. Roder’s “The East” (2
It was found that it belongs to the genus Candida, which is described in the 1970 edition. Species identification Lee & Komagata (1980)
“Taxonomical study of methanol−
According to the genus Candeida, there are five species of methanol-assimilating bacteria (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 boidnii
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 various properties of this strain (A-1917-3) were candeida
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. Moreover, 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 4-10, the temperature is 20-
Selected from around 40℃. In the production of methyl formate, both proliferating bacterial cells and dormant bacterial cells can be used. When collecting and purifying methyl formate 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. Candida boideinii A-1917 isolated from natural soil
- Slant culture of 3 bacteria (30℃, 3-6 days) A loopful of the bacteria was inoculated into the above Colben, cultured for 3 days at 30℃ using a reciprocating shaker at 112 cycles/min, and 50ml of this culture was grown. 2 mg of methyl formate was obtained.
Claims (1)
ル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、培養物からギ酸
メチルを得ることを特徴とするギ酸メチルの製造
方法。1. A method for producing methyl formate, which comprises culturing a microorganism belonging to the genus Candida and having the ability to produce methyl formate using methanol as a carbon source, and obtaining methyl formate from the culture.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP21424884A JPS6192584A (en) | 1984-10-15 | 1984-10-15 | Production of methyl formate |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP21424884A JPS6192584A (en) | 1984-10-15 | 1984-10-15 | Production of methyl formate |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPS6192584A JPS6192584A (en) | 1986-05-10 |
| JPH0424989B2 true JPH0424989B2 (en) | 1992-04-28 |
Family
ID=16652621
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP21424884A Granted JPS6192584A (en) | 1984-10-15 | 1984-10-15 | Production of methyl formate |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS6192584A (en) |
-
1984
- 1984-10-15 JP JP21424884A patent/JPS6192584A/en active Granted
Also Published As
| Publication number | Publication date |
|---|---|
| JPS6192584A (en) | 1986-05-10 |
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Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| EXPY | Cancellation because of completion of term |