| 产异丁醇关键基因在大肠杆菌中表达的研究 |
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| 引用本文： | 潘超强,高强,郑春阳,孟庆艳,段强,冯少龙.产异丁醇关键基因在大肠杆菌中表达的研究[J].微生物学通报,2012,39(7):0912-0920. |
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| 作者姓名： | 潘超强 高强 郑春阳 孟庆艳 段强 冯少龙 |
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| 作者单位： | 1. 天津科技大学生物工程学院工业发酵微生物教育部重点实验室工业酶国家工程实验室 天津 300457
2. 天津强微特生物科技有限公司 天津 300384 |
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| 基金项目： | 国家973计划项目(No. 2007CB714305, No. 2011CB707401); 国家863计划项目(No. 2012AA021302); 天津市滨海新区自主创新重大项目(No. 2011-BK120014) |
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| 摘 要： | 目的]改造大肠杆菌缬氨酸合成途径,使其能够代谢合成异丁醇.方法]将乳酸乳球菌(Lactococcus lactis) 1.2829的2-酮异戊酸脱羧酶基因(kivD)和醇脱氢酶基因(adhA)串联克隆到大肠杆菌DH5α宿主中表达.结果]经过改造的宿主菌发酵24 h后异丁醇产量为0.12 g/L.酶活测定实验发现,kivD和adhA基因在宿主菌中均得到表达,但由于KivD的低表达量导致宿主菌最终的异丁醇合成能力偏低.通过研究温度和pH对KivD和AdhA酶活的影响,最终选定二者的最适温度为30℃,最适pH为6.5. 结论]通过向宿主菌导入外源异丁醇合成基因能够改造其自身代谢途径,从而合成异丁醇.
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| 关 键 词： | 2-酮异戊酸脱羧酶 醇脱氢酶 大肠杆菌DH5α 异丁醇 |
Coexpression of two essential isobutanol synthesis genes in Escherichia coli |
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| PAN Chao-Qiang,GAO Qiang,ZHENG Chun-Yang,MENG Qing-Yan,DUAN Qiang and FENG Shao-Long.Coexpression of two essential isobutanol synthesis genes in Escherichia coli[J].Microbiology,2012,39(7):0912-0920. |
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| Authors: | PAN Chao-Qiang GAO Qiang ZHENG Chun-Yang MENG Qing-Yan DUAN Qiang and FENG Shao-Long |
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| Institution: | 1. Key Laboratory of Industrial Fermentation Microbiology, Ministry of Education, College of Biotechnology, Tianjin University of Science & Technology, National Engineering Laboratory for Industry Enzymes, Tianjin 300457, China;2. Robustnique Corporation Ltd., Tianjin 300384, China;1. Key Laboratory of Industrial Fermentation Microbiology, Ministry of Education, College of Biotechnology, Tianjin University of Science & Technology, National Engineering Laboratory for Industry Enzymes, Tianjin 300457, China |
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| Abstract: | Objective] E.coli DH5α modified the valine biosynthesis pathway to biosynthesize isobutanol.Methods] The 2-ketoisovalerate decarboxylase gene(kivD) and alcohol dehydro-genase gene(adhA) of Lactococcus lactis 1.2829 were tandemly cloned and expressed in E.coli DH5α.Results] The yield of isobatanol by the engineered E.coli was only 0.12 g/L by 24 h fermentation.Further results revealed that the insufficient KivD activity is the bottleneck for the isobutanol biosynthesis.A series of experiments also showed that the optimal tempera-ture and pH for both KivD and AdhA are 30 °C and pH 6.5,respectively.Conclusion] Iso-batanol fermentation by cloning and expressing essential genes in host is feasible. |
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| Keywords: | 2-Ketoisovalerate decarboxylase Alcohol dehydrogenase E coli DH5α Isobutanol |
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