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林丽华,郭媛,裴建新,庞浩,黄日波.产丁醇大肠杆菌工程菌的构建[J].广西科学,2014,21(1):42-46. [点击复制]
- LIN Li-hua,GUO Yuan,PEI Jian-xin,PANG Hao,HUANG Ri-bo.The Construction of Recombinant Escherichia coli Producing Butanol[J].Guangxi Sciences,2014,21(1):42-46. [点击复制]
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| 产丁醇大肠杆菌工程菌的构建 |
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林丽华1,2, 郭媛1, 裴建新1, 庞浩1,2, 黄日波1,2
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| (1.广西科学院, 非粮生物质酶解国家重点实验室, 国家非粮生物质能源工程技术研究中心, 广西生物质产业化工程院, 广西生物炼制重点实验室, 广西南宁 530007;2.广西大学生命科学与技术学院, 广西南宁 530004) |
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| 摘要: |
| [目的]为了在大肠杆菌(Escherichia coli)中导入改良的丁醇合成途径,使非生产菌株大肠杆菌具备产丁醇的能力。[方法]克隆大肠杆菌乙酰转移酶基因atoB和丙酮丁醇梭菌(Clostridium acetobutylicum)丁醇合成途径关键酶基因(crt、hbd、adhE),构建多顺反子表达质粒pSE380-atoB-adhE-crt-hbd;克隆齿垢密螺旋体(Treponema denticola)反式烯酰辅酶A还原酶基因ter,构建表达质粒pSTV29-ter,并将双质粒导入到大肠杆菌。[结果]构建的工程菌能半厌氧发酵产微量丁醇,产量为0.08g/L。[结论]大肠杆菌中的丁醇合成途径导入成功,构建了产丁醇的大肠杆菌工程菌。 |
| 关键词: 正丁醇 大肠杆菌 丙酮丁醇梭菌 齿垢密螺旋体 半厌氧发酵 |
| DOI： |
| 投稿时间：2013-08-05修订日期：2013-08-26 |
| 基金项目:广西科技合作与交流计划项目(桂科合1347004-1),广西科技攻关项目(桂科攻10123007-3),广西科学院基本科研业务费项目(12YJ25SW05、13YJ22SW02),国家自然科学基金(21366007)资助。 |
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| The Construction of Recombinant Escherichia coli Producing Butanol |
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LIN Li-hua1,2, GUO Yuan1, PEI Jian-xin1, PANG Hao1,2, HUANG Ri-bo1,2
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| (1.Guangxi Academy of Sciences, State Key Laboratory of Non-Food Biomass and Enzyme Technology, National Engineering Research Center for Non-food Biorefinery, Guangxi Biomass Industrialization Engineering Institute, Guangxi Key Laboratory of Biorefinery, Nanning, Guangxi, 530007, China;2.College of Life Science & Technology of Guangxi University) |
| Abstract: |
| [Objective] A modified 1-butanol pathway was constructed in non-native producer Escherichia coli to produce butanol.[Method] By cloning the acetyltransferase gene from E.coli, and the key butanol synthetic pathway genes crt, hbd and adhE from Clostridium acetobutylicum, the polycistron expression plasmid pSE380-atoB-adhE-crt-hbd was constructed;by cloning the trans-2-enoyl-CoA reductase gene ter from Treponema denticola, the expression plasmid pSTV29-ter was constructed.[Result] The recombinant E.coli containing the double plasmids was fermented in micro-aerobic and the maximal concentration of butanol was 0.08g/L at 24h.[Conclusion] The butanol synthetic pathway was expressed in E.coli and the recombinant strain of producing butanol was constructed successfully. |
| Key words: 1-butanol Escherichia coli Clostridium acetobutylicum Treponema denticola micro-aerobic fermentation |
