Global gene expression analysis of glucose overflow metabolism in Escherichia coli and reduction of aerobic acetate formation
Global gene expression analysis of glucose overflow metabolism in Escherichia coli and reduction of aerobic acetate formation
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DOI：
10.1007/s00253-006-0680-3
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发表时间：
2007-02-01
影响因子：
5
通讯作者：
Wendisch, Volker F.
中科院分区：
文献类型：
--
作者：
Veit, Andrea;Polen, Tino;Wendisch, Volker F.
在葡萄糖的有氧生长过程中，大肠杆菌在所谓的溢出代谢中产生醋酸盐。采用DNA微阵列分析，确定了大肠杆菌MG1655在葡萄糖有氧生长过程中以不同醋酸酯形成速率为特征的趋化培养物的整体基因表达模式。相关分析发现，分别编码TCA循环酶琥珀酸脱氢酶、α -酮戊二酸脱氢酶、琥珀酰辅酶A合成酶、乌头酸酶、富马酸酶和苹果酸脱氢酶的10个基因（sdhCDAB、sub、suc、acnB、lpdA、fumC和mdh）和乙酸利用操纵子acs-yjcH-actP基因的表达与乙酸形成呈负相关。通过染色体启动子交换诱变解除sdhCDAB-b0725-sucABCD操纵子的转录控制，获得了由该操纵子编码的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的菌株。与亲本菌株MG1655相比，该菌株在保持高生长和葡萄糖消耗率的同时产生更少的乙酸，并将更多的碳用于二氧化碳的形成。
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.