JPS63102687A - Production of butanol - Google Patents
Production of butanolInfo
- Publication number
- JPS63102687A JPS63102687A JP24737286A JP24737286A JPS63102687A JP S63102687 A JPS63102687 A JP S63102687A JP 24737286 A JP24737286 A JP 24737286A JP 24737286 A JP24737286 A JP 24737286A JP S63102687 A JPS63102687 A JP S63102687A
- Authority
- JP
- Japan
- Prior art keywords
- butanol
- alcohol
- culture
- ferm
- extractant
- 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
Links
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 title claims abstract description 54
- 238000004519 manufacturing process Methods 0.000 title claims description 8
- 238000000034 method Methods 0.000 claims abstract description 10
- 150000001298 alcohols Chemical class 0.000 claims abstract description 9
- 241000193446 Thermoanaerobacterium thermosaccharolyticum Species 0.000 claims abstract description 7
- 238000000605 extraction Methods 0.000 claims abstract description 5
- 230000003698 anagen phase Effects 0.000 claims abstract description 3
- 125000004432 carbon atom Chemical group C* 0.000 claims description 5
- 241001148471 unidentified anaerobic bacterium Species 0.000 claims description 4
- 235000000346 sugar Nutrition 0.000 claims description 2
- 150000008163 sugars Chemical class 0.000 claims description 2
- GLDOVTGHNKAZLK-UHFFFAOYSA-N octadecan-1-ol Chemical compound CCCCCCCCCCCCCCCCCCO GLDOVTGHNKAZLK-UHFFFAOYSA-N 0.000 abstract description 18
- GOQYKNQRPGWPLP-UHFFFAOYSA-N heptadecan-1-ol Chemical compound CCCCCCCCCCCCCCCCCO GOQYKNQRPGWPLP-UHFFFAOYSA-N 0.000 abstract description 13
- 241000894006 Bacteria Species 0.000 abstract description 12
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 abstract description 6
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 abstract description 6
- BXWNKGSJHAJOGX-UHFFFAOYSA-N hexadecan-1-ol Chemical compound CCCCCCCCCCCCCCCCO BXWNKGSJHAJOGX-UHFFFAOYSA-N 0.000 abstract description 6
- 229910052799 carbon Inorganic materials 0.000 abstract description 5
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 abstract description 4
- 241000193403 Clostridium Species 0.000 abstract description 3
- 150000001720 carbohydrates Chemical class 0.000 abstract description 3
- 229960000541 cetyl alcohol Drugs 0.000 abstract description 3
- 229910052757 nitrogen Inorganic materials 0.000 abstract description 3
- 229910017053 inorganic salt Inorganic materials 0.000 abstract 1
- 230000000050 nutritive effect Effects 0.000 abstract 1
- 238000000855 fermentation Methods 0.000 description 10
- 230000004151 fermentation Effects 0.000 description 10
- 239000002609 medium Substances 0.000 description 4
- 244000005700 microbiome Species 0.000 description 4
- 230000000052 comparative effect Effects 0.000 description 3
- 238000005192 partition Methods 0.000 description 3
- SRBFZHDQGSBBOR-IOVATXLUSA-N D-xylopyranose Chemical compound O[C@@H]1COC(O)[C@H](O)[C@H]1O SRBFZHDQGSBBOR-IOVATXLUSA-N 0.000 description 2
- 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 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- 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 2
- 238000011109 contamination Methods 0.000 description 2
- 239000008103 glucose Substances 0.000 description 2
- HLZKNKRTKFSKGZ-UHFFFAOYSA-N tetradecan-1-ol Chemical compound CCCCCCCCCCCCCCO HLZKNKRTKFSKGZ-UHFFFAOYSA-N 0.000 description 2
- 231100000331 toxic Toxicity 0.000 description 2
- 230000002588 toxic effect Effects 0.000 description 2
- ALSTYHKOOCGGFT-KTKRTIGZSA-N (9Z)-octadecen-1-ol Chemical compound CCCCCCCC\C=C/CCCCCCCCO ALSTYHKOOCGGFT-KTKRTIGZSA-N 0.000 description 1
- OWEGMIWEEQEYGQ-UHFFFAOYSA-N 100676-05-9 Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC2C(OC(O)C(O)C2O)CO)O1 OWEGMIWEEQEYGQ-UHFFFAOYSA-N 0.000 description 1
- GUBGYTABKSRVRQ-XLOQQCSPSA-N Alpha-Lactose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@H](O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-XLOQQCSPSA-N 0.000 description 1
- GUBGYTABKSRVRQ-CUHNMECISA-N D-Cellobiose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-CUHNMECISA-N 0.000 description 1
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 description 1
- GUBGYTABKSRVRQ-PICCSMPSSA-N Maltose Natural products O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-PICCSMPSSA-N 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- PYMYPHUHKUWMLA-UHFFFAOYSA-N arabinose Natural products OCC(O)C(O)C(O)C=O PYMYPHUHKUWMLA-UHFFFAOYSA-N 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- SRBFZHDQGSBBOR-UHFFFAOYSA-N beta-D-Pyranose-Lyxose Natural products OC1COC(O)C(O)C1O SRBFZHDQGSBBOR-UHFFFAOYSA-N 0.000 description 1
- GUBGYTABKSRVRQ-QUYVBRFLSA-N beta-maltose Chemical compound OC[C@H]1O[C@H](O[C@H]2[C@H](O)[C@@H](O)[C@H](O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@@H]1O GUBGYTABKSRVRQ-QUYVBRFLSA-N 0.000 description 1
- ALSTYHKOOCGGFT-UHFFFAOYSA-N cis-oleyl alcohol Natural products CCCCCCCCC=CCCCCCCCCO ALSTYHKOOCGGFT-UHFFFAOYSA-N 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 239000000446 fuel Substances 0.000 description 1
- 238000004817 gas chromatography Methods 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 239000008101 lactose Substances 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 239000013028 medium composition Substances 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 229920006395 saturated elastomer Polymers 0.000 description 1
- 239000002689 soil Substances 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
- 229920001221 xylan Polymers 0.000 description 1
- 150000004823 xylans Chemical class 0.000 description 1
Abstract
Description
【発明の詳細な説明】
〔産業上の利用分野〕
本発明はブタノールの製造方法に関し、詳しくは好熱嫌
気性菌を用いて抽出発酵法によりブタノールを製造する
にあたり、抽剤として特定のアルコールを使用すること
によってブタノールを効率よく生産する方法に関する。[Detailed Description of the Invention] [Industrial Application Field] The present invention relates to a method for producing butanol, and more specifically, in producing butanol by an extractive fermentation method using thermophilic anaerobic bacteria, a specific alcohol is used as an extractant. This invention relates to a method for efficiently producing butanol.
〔従来技術及び発明が解決しようとする問題点〕発酵法
によるブタノールの製造においては、生産物であるブタ
ノールが微生物に対して毒性を持ツタメ、ブタノ−少を
高濃度に生産することができない。[Prior Art and Problems to be Solved by the Invention] In the production of butanol by the fermentation method, the product, butanol, is toxic to microorganisms, and it is not possible to produce a high concentration of butanol.
このよう−゛発酵生産物による微生物への阻害を回避す
る手段として、抽出培養が知られている。Extraction culture is known as a means of avoiding such inhibition of microorganisms by fermentation products.
ブタノールの発酵生産を抽出培養により行なう場合の抽
剤としては、炭素数１０〜１４個のアルコール（特開昭
５９−２１６５９１号）、炭素数１６〜１８個の不飽和
アルコールまたは炭素数１６〜２０個の側鎖を有するア
ルコール（特開昭６０＝１７２２９０号）が知られてい
る。When fermentation production of butanol is carried out by extraction culture, extractants include alcohols having 10 to 14 carbon atoms (JP-A-59-216591), unsaturated alcohols having 16 to 18 carbon atoms, or unsaturated alcohols having 16 to 20 carbon atoms. Alcohols having side chains (JP-A-60-172290) are known.
しかしながら、これら抽剤を使用するブタノール発酵は
、中温菌を用いる場合に適用されるのであり、高温下で
ブタノール発酵を行なう好熱菌に対しては、これら抽剤
が該好熱菌に対して毒性を示すという問題点がある。However, butanol fermentation using these extractants is applied when mesophilic bacteria are used, and these extractants are effective against thermophilic bacteria that carry out butanol fermentation at high temperatures. The problem is that it is toxic.
一方、雑菌汚染の防止、ブタノール回収コストの低減等
の立場から高温発酵によってブタノールを製造する方法
が望まれている。On the other hand, a method for producing butanol by high-temperature fermentation is desired from the standpoint of preventing bacterial contamination and reducing butanol recovery costs.
本発明者は好熱菌を用いて高温発酵によりブタノールを
製造する場合の抽剤について検討し、特定のアルコール
を使用すれば好熱菌に毒性を示さないことを見出し、本
発明に到達した。The present inventor studied extractants for producing butanol by high-temperature fermentation using thermophilic bacteria, and found that the use of a specific alcohol does not show toxicity to thermophilic bacteria, and has arrived at the present invention.
本発明は好熱嫌気性菌を用いてＰｉ類からブタノールを
抽出培養によって製造する方法において、抽剤として炭
素数１５〜１８個の直鎖飽和アルコールを使用すること
を特徴とするブタノールの製造方法である。The present invention is a method for producing butanol from Pi by extraction culture using thermophilic anaerobic bacteria, which is characterized in that a linear saturated alcohol having 15 to 18 carbon atoms is used as an extractant. It is.
本発明に用いる好熱嫌気性菌としては高温（通常は４５
〜６５℃）において糖類からブタノールを生産する能力
を有するものであればよく、クロストリジウム属に属す
る好熱菌が好ましく、特にクロストリジウム・サーモサ
ッカロリティカムが好適である。このようなりロストリ
ジウム属の微生物として例えば本発明者らが土壌より分
離したクロストリジウム・サーモサフカロリティヵムＢ
−２５８株（ＦＥＲＭ Ｐ−８２７３）、クロストリ
ジウム・サーモサッカロリティカムＣＢ−１６６６株（
ＦＥＲＭ Ｐ−８２７４）、クロストリジウム・サー
モサッカロリティカム６９５７株（ＦＥＲＭ Ｐ−８
０７１）、 クロストリジウム・サーモサッカロリテ
ィカム ＡＴＣＣ７９５６株等が挙げられる。The thermophilic anaerobic bacteria used in the present invention are used at high temperatures (usually 45%
Any bacteria can be used as long as it has the ability to produce butanol from saccharides at a temperature of 65° C. to 65° C., and thermophilic bacteria belonging to the genus Clostridium are preferred, with Clostridium thermosaccharolyticum being particularly preferred. As a microorganism belonging to the genus Rostridia, for example, Clostridium thermosafcalolyticum B, which the present inventors isolated from soil,
-258 strain (FERM P-8273), Clostridium thermosaccharolyticum CB-1666 strain (
FERM P-8274), Clostridium thermosaccharolyticum strain 6957 (FERM P-8
071), Clostridium thermosaccharolyticum ATCC 7956 strain, and the like.
培養に用いる培地は炭素源、窒素源、無機塩類。The culture medium used is a carbon source, nitrogen source, and inorganic salts.
有機栄養素などを含有する通常の培地であり、炭素源と
してはグルコース、セロビオース、シュクロース、ラク
トース、キシロース、マルトース。It is a normal medium containing organic nutrients, etc., and the carbon sources are glucose, cellobiose, sucrose, lactose, xylose, and maltose.
キシラン、デンプン等の糖類が使用される。窒素源等は
微生物が良好に生育してブタノールを効率よく生産しう
るちのを適宜選択すればよい。Sugars such as xylan and starch are used. The nitrogen source may be appropriately selected from a source that allows microorganisms to grow well and efficiently produce butanol.
さらに、本発明では培養にあたり抽剤として炭素数１５
〜１８個の直鎖飽和アルコールを使用する。これらアル
コールのうちではセチルアルコール、ヘプタデシルアル
コール、ステアリルアルコールが好適であり、これらを
単独でもしくは２種以上を組合せて使用する。抽剤とし
てのアルコールの添加量は培養液量の２０〜１００％が
適当であり、培養開始時から微生物の対数増殖期中期（
ブタノールが５ ｇ／ｌ程度蓄積したとき）までの間に
全量を１回で、もしくは複数回に分割して加える。Furthermore, in the present invention, a carbon number of 15 is used as an extractant for culturing.
~18 straight chain saturated alcohols are used. Among these alcohols, cetyl alcohol, heptadecyl alcohol, and stearyl alcohol are preferred, and these may be used alone or in combination of two or more. The appropriate amount of alcohol to be added as an extractant is 20 to 100% of the culture solution volume, from the start of culture to the mid-logarithmic growth phase (
Add the entire amount at once or in multiple portions until the amount of butanol has accumulated to about 5 g/l.
好熱菌の培養条件は使用する菌株によって異なるが、通
常４０〜６５℃、好ましくは５５〜６０”Ｃ，Ｉ）Ｈ５
〜８、好ましくは５．５〜７に調節して嫌気的条件下で
行なう。培養期間については、目的トするアルコール類
が十分量生産されるまで行なえばよく、通常は１〜１０
日間、好ましくは２〜５日間である。Culture conditions for thermophilic bacteria vary depending on the strain used, but are usually 40-65°C, preferably 55-60"C, I) H5
-8, preferably 5.5-7, and carried out under anaerobic conditions. Regarding the culture period, it is sufficient to carry out the culture until a sufficient amount of the desired alcohol is produced, which is usually 1 to 10 hours.
2 to 5 days, preferably 2 to 5 days.
次に、本発明を実施例により説明するが、本発明はこれ
らによって制限されるものではない。Next, the present invention will be explained by examples, but the present invention is not limited thereto.
実施例１
第１表に示した培地■にクロストリジウム・サーモサッ
カロリティカム Ｂ−２５８株（ＦＥＲＭＰ−８２７３
）を接種し、６０℃で１８時間嫌気的に液体培養した。Example 1 Clostridium thermosaccharolyticum strain B-258 (FERMP-8273
) was inoculated and cultured in liquid form anaerobically at 60°C for 18 hours.
この培養液を第１表に示した培地１１１０ｍ／！に２％
の割合で接種し、ステアリルアルコール（分配係数５．
５）２．５ｇを加えて５０ｍ１！ネジロ試験管中で６０
℃にて５日間嫌気的に振とう培養した。This culture solution was used in the medium shown in Table 1, 1110 m/! 2% to
Stearyl alcohol (partition coefficient 5.
5) Add 2.5g and make 50ml! 60 in Nejiro test tube
The culture was carried out anaerobically with shaking at ℃ for 5 days.
得られた培養液を遠心分離して除菌後、リン酸（３３ｍ
ｇ／ｍｆｆ）で酸性にし、ガスクロマトグラフにより生
産物を分析した。ブタノールの全生産量はブタノールの
各アルコールと水中の分配係数から計算により求めた。The obtained culture solution was centrifuged to remove bacteria, and then phosphoric acid (33 m
g/mff) and the product was analyzed by gas chromatography. The total production of butanol was calculated from the partition coefficient of butanol between each alcohol and water.
結果を第２表に示す。The results are shown in Table 2.
実施例２
実施例１においてステアリルアルコールの代すにセチル
アルコール（分配係数４．５）５ｇを加えたこと以外は
実施例１と同様に行なった。結果を第２表に示す。Example 2 The same procedure as in Example 1 was carried out except that 5 g of cetyl alcohol (partition coefficient 4.5) was added instead of stearyl alcohol. The results are shown in Table 2.
実施例３
実施例１において培地Ｈの代りに培地■を用い、ステア
リルアルコールを加えずに培養を開始し、２日後にグル
コース０．５ｇとステアリルアルコ−ル
行なった。結果を第２表に示す。Example 3 In Example 1, medium 1 was used instead of medium H, culture was started without adding stearyl alcohol, and 2 days later, 0.5 g of glucose and stearyl alcohol were added. The results are shown in Table 2.
比較例１
）Ｊｌ［においてステアリルアルコールを加えずに培養
したこと以外は実施例１と同様に行なった。結果を第２
表に示す。Comparative Example 1) The same procedure as in Example 1 was carried out except that the culture was carried out in Jl without adding stearyl alcohol. Second result
Shown in the table.
比較例２
実施例１においてステアリルアルコールの７１にオレイ
ンアルコールを用いたこと以外は実施例１と同様に行な
った。結果を第２表に示す。Comparative Example 2 The same procedure as in Example 1 was conducted except that oleic alcohol was used as 71 of the stearyl alcohol in Example 1. The results are shown in Table 2.
比較例３
実施例１においてステアリルアルコールの代りにテトラ
デカノールを用いたこと以外は実施例１と同様に行なっ
た。結果を第２表に示す。Comparative Example 3 The same procedure as in Example 1 was conducted except that tetradecanol was used instead of stearyl alcohol. The results are shown in Table 2.
１、／
第１表（培地組成）
〔発明の効果〕
本発明によれば、抽剤を用いる高温発酵によってブタノ
ールを効率よく製造することができる。1./Table 1 (Medium Composition) [Effects of the Invention] According to the present invention, butanol can be efficiently produced by high temperature fermentation using an extractant.
高温で発酵を行なうため、ブタノールを経済的に回収す
ることができ、発酵槽の冷却コストの低減、雑菌による
汚染の防止等を図ることができる。また、ブタノールは
工業用、燃料用等として利用される。Since fermentation is carried out at a high temperature, butanol can be recovered economically, the cost of cooling the fermenter can be reduced, and contamination by various bacteria can be prevented. In addition, butanol is used for industrial purposes, fuel, etc.
Claims (3)
培養によって製造する方法において、抽剤として炭素数
１５〜１８個の直鎖飽和アルコールを使用することを特
徴とするブタノールの製造方法。(1) A method for producing butanol from sugars by extraction culture using thermophilic anaerobic bacteria, which is characterized in that a linear saturated alcohol having 15 to 18 carbon atoms is used as an extractant.
ロリティカムである特許請求の範囲第１項記載の方法。(2) The method according to claim 1, wherein the thermophilic anaerobe is Clostridium thermosaccharolyticum.
添加する特許請求の範囲第１項記載の方法。(3) The method according to claim 1, wherein the extractant is added between the start of culture and the middle of the logarithmic growth phase.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP24737286A JPS63102687A (en) | 1986-10-20 | 1986-10-20 | Production of butanol |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP24737286A JPS63102687A (en) | 1986-10-20 | 1986-10-20 | Production of butanol |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPS63102687A true JPS63102687A (en) | 1988-05-07 |
| JPH044871B2 JPH044871B2 (en) | 1992-01-29 |
Family
ID=17162449
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP24737286A Granted JPS63102687A (en) | 1986-10-20 | 1986-10-20 | Production of butanol |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS63102687A (en) |
Cited By (15)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US7541173B2 (en) | 2006-06-15 | 2009-06-02 | E.I. Du Pont De Nemours And Company | Solvent tolerant microorganisms and methods of isolation |
| US7659104B2 (en) | 2006-05-05 | 2010-02-09 | E.I. Du Pont De Nemours And Company | Solvent tolerant microorganisms and methods of isolation |
| JP2010161990A (en) * | 2009-01-16 | 2010-07-29 | Nippon Shokubai Co Ltd | Method for sterilizing culture medium and method for producing 1-butanol by fermentation |
| US7851188B2 (en) | 2005-10-26 | 2010-12-14 | Butamax(Tm) Advanced Biofuels Llc | Fermentive production of four carbon alcohols |
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| US8236994B2 (en) | 2006-10-31 | 2012-08-07 | Metabolic Explorer | Process for the biological production of 1,3-propanediol from glycerol with high yield |
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-
1986
- 1986-10-20 JP JP24737286A patent/JPS63102687A/en active Granted
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| Publication number | Publication date |
|---|---|
| JPH044871B2 (en) | 1992-01-29 |
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