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学科主题微生物学
Construction of E. coli NAD+ auxotrophic strains and the biotechnological application thereof
Zhou YJ(周雍进); Hou SH(侯淑华); Zhang SF(张素芳); Zhao ZB(赵宗保)
会议文集Book of Abstracts
会议名称Asian Congress on Biotechnology
会议日期2011-5-11
2011
会议地点上海
页码26-0
出版者待补充
出版地待补充
合作性质分会口头报告
部门归属1816
主办者亚洲生物技术联合会
英文摘要NAD+ and its reduced form NADH are essential cofactors in biological systems. They function as cofactors in over 300 redox reactions in vivo1. The level of NAD+ and NADH is tightly controlled by a variety of mechanisms including their biosynthesis and salvage. Therefore, it is difficult to answer some fundamental questions such as the minimal NAD+ level for cell growth and the biological consequences of abnormal activity of a specific NAD+-dependent enzyme. We expressed the NTT4 gene from Chlamydiae UWE25 in Escherichia coli BW25113, as the NTT4 protein was reported as a NAD+ transporter that can specifically transport intact NAD+ across cytoplasmic membrane2. We knocked out the nadC gene responsible for de novo biosynthesis of NAD+ and constructed the strain E. coli BW25113 (ΔnadC, NTT4). It was found that NAD+ in the culture media could significantly promote the growth of BW25113 (ΔnadC, NTT4), suggesting that the NTT4 protein was functional in E. coli (Fig A, B). We then disrupted the other two genes, nadD and nadE, and obtained the strains E. coli BW25113 (ΔnadD, NTT4) and BW25113 (ΔnadE, NTT4). Cell growth of these two strains are depending on exogenous NAD+ supplemented in the media, suggesting that we have successfully engineered E. coli to hold an NAD+ auxotrophic phenotype. We are carrying out a number of experiments using these NAD+ auxotrophic strains to address some interesting questions which may not be able to do otherwise. Results will be discussed during the conference.; NAD+ and its reduced form NADH are essential cofactors in biological systems. They function as cofactors in over 300 redox reactions in vivo1. The level of NAD+ and NADH is tightly controlled by a variety of mechanisms including their biosynthesis and salvage. Therefore, it is difficult to answer some fundamental questions such as the minimal NAD+ level for cell growth and the biological consequences of abnormal activity of a specific NAD+-dependent enzyme. We expressed the NTT4 gene from Chlamydiae UWE25 in Escherichia coli BW25113, as the NTT4 protein was reported as a NAD+ transporter that can specifically transport intact NAD+ across cytoplasmic membrane2. We knocked out the nadC gene responsible for de novo biosynthesis of NAD+ and constructed the strain E. coli BW25113 (ΔnadC, NTT4). It was found that NAD+ in the culture media could significantly promote the growth of BW25113 (ΔnadC, NTT4), suggesting that the NTT4 protein was functional in E. coli (Fig A, B). We then disrupted the other two genes, nadD and nadE, and obtained the strains E. coli BW25113 (ΔnadD, NTT4) and BW25113 (ΔnadE, NTT4). Cell growth of these two strains are depending on exogenous NAD+ supplemented in the media, suggesting that we have successfully engineered E. coli to hold an NAD+ auxotrophic phenotype. We are carrying out a number of experiments using these NAD+ auxotrophic strains to address some interesting questions which may not be able to do otherwise. Results will be discussed during the conference.
语种英语
文献类型会议论文
条目标识符http://cas-ir.dicp.ac.cn/handle/321008/116038
专题中国科学院大连化学物理研究所
通讯作者Zhao ZB(赵宗保)
推荐引用方式
GB/T 7714
Zhou YJ,Hou SH,Zhang SF,et al. Construction of E. coli NAD+ auxotrophic strains and the biotechnological application thereof[C]. 待补充:待补充,2011:26-0.
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