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大肠杆菌中葡萄糖溢流代谢的全基因组表达分析及需氧乙酸生成的减少

Global gene expression analysis of glucose overflow metabolism in Escherichia coli and reduction of aerobic acetate formation.

作者信息

Veit Andrea, Polen Tino, Wendisch Volker F

机构信息

Institute of Biotechnology 1, Research Center Juelich, 52428 Juelich, Germany.

出版信息

Appl Microbiol Biotechnol. 2007 Feb;74(2):406-21. doi: 10.1007/s00253-006-0680-3. Epub 2006 Nov 25.

Abstract

During aerobic growth on glucose, Escherichia coli produces acetate in the so-called overflow metabolism. DNA microarray analysis was used to determine the global gene expression patterns of chemostat cultivations of E. coli MG1655 that were characterized by different acetate formation rates during aerobic growth on glucose. A correlation analysis identified that expression of ten genes (sdhCDAB, sucB, sucC, acnB, lpdA, fumC and mdh) encoding the TCA cycle enzymes succinate dehydrogenase, alpha-ketoglutarate dehydrogenase, succinyl-CoA synthetase, aconitase, fumarase and malate dehydrogenase, respectively, and of the acs-yjcH-actP operon for acetate utilization correlated negatively with acetate formation. Relieving transcriptional control of the sdhCDAB-b0725-sucABCD operon by chromosomal promoter exchange mutagenesis yielded a strain with increased specific activities of the TCA cycle enzymes succinate dehydrogenase, alpha-ketoglutarate dehydrogenase and succinyl-CoA synthetase, which are encoded by this operon. The resulting strain produced less acetate and directed more carbon towards carbon dioxide formation than the parent strain MG1655 while maintaining high growth and glucose consumption rates.

摘要

在以葡萄糖为底物进行有氧生长期间,大肠杆菌在所谓的溢流代谢过程中产生乙酸盐。利用DNA微阵列分析来确定大肠杆菌MG1655恒化培养物的全局基因表达模式,这些培养物的特征是在以葡萄糖为底物进行有氧生长期间具有不同的乙酸盐形成速率。相关性分析确定,分别编码三羧酸循环(TCA)酶琥珀酸脱氢酶、α-酮戊二酸脱氢酶、琥珀酰辅酶A合成酶、乌头酸酶、延胡索酸酶和苹果酸脱氢酶的十个基因(sdhCDAB、sucB、sucC、acnB、lpdA、fumC和mdh)以及用于乙酸盐利用的acs-yjcH-actP操纵子的表达与乙酸盐形成呈负相关。通过染色体启动子交换诱变解除对sdhCDAB-b0725-sucABCD操纵子的转录控制,得到了一种菌株,该菌株中由该操纵子编码的TCA循环酶琥珀酸脱氢酶、α-酮戊二酸脱氢酶和琥珀酰辅酶A合成酶的比活性增加。与亲本菌株MG1655相比,所得菌株产生的乙酸盐更少,并且将更多的碳导向二氧化碳的形成,同时保持高生长速率和葡萄糖消耗速率。

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