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蛋白质N(ε)-赖氨酸乙酰化对大肠杆菌BL21中乙酸代谢的调控

Regulation of acetate metabolism in Escherichia coli BL21 by protein N(ε)-lysine acetylation.

作者信息

Castaño-Cerezo Sara, Bernal Vicente, Röhrig Teresa, Termeer Svenja, Cánovas Manuel

机构信息

Departamento de Bioquímica y Biología Molecular B e Inmunología, Facultad de Química, Campus Regional de Excelencia "Mare Nostrum", Murcia, Spain,

出版信息

Appl Microbiol Biotechnol. 2015 Apr;99(8):3533-45. doi: 10.1007/s00253-014-6280-8. Epub 2014 Dec 20.

Abstract

Acetate production is one of the most striking differences between Escherichia coli K12 and BL21 strains. Transcription of acetate metabolism genes is regulated. Additionally, acetyl-CoA synthetase, which activates acetate to acetyl-CoA, is regulated by post-translational acetylation. The aim of this study was to understand the contribution of reversible protein lysine acetylation to the regulation of acetate metabolism in E. coli BL21. The phenotypic differences between both strains were especially important in the presence of acetate. The high expression of acetyl-CoA synthetase (acs) in glucose exponential phase in BL21 allows the simultaneous consumption of acetate and glucose. Lack of catabolite repression also affected its post-translational regulator, the protein acetyltransferase (patZ). The effect of the deletion of cobB (encoding a sirtuin-like protein deacetylase) and patZ genes depended on the genetic background. The deletion of cobB in both strains increased acetate production and decreased growth rate in acetate cultures. The deletion of patZ in BL21 suppressed acetate overflow in glucose medium and increased the growth rate in acetate cultures. Differences on acetate overflow between BL21 and K12 strains are caused by many overlapping factors. Two major contributing effects were identified: (1) the expression of acs during exponential growth is not repressed in the BL21 strain due to concomitant cAMP production and (2) the acetyl-CoA synthetase activity is more tightly regulated by protein acetylation in BL21 than in the K12. Altogether these differences contribute to the lower acetate overflow and the improved ability of E. coli BL21 to consume this metabolite in the presence of glucose.

摘要

乙酸盐的产生是大肠杆菌K12和BL21菌株之间最显著的差异之一。乙酸盐代谢基因的转录受到调控。此外,将乙酸盐激活为乙酰辅酶A的乙酰辅酶A合成酶受到翻译后乙酰化的调控。本研究的目的是了解可逆蛋白赖氨酸乙酰化对大肠杆菌BL21中乙酸盐代谢调控的作用。在乙酸盐存在的情况下,两种菌株之间的表型差异尤为重要。BL21中葡萄糖指数生长期乙酰辅酶A合成酶(acs)的高表达使得乙酸盐和葡萄糖能够同时被消耗。缺乏分解代谢物阻遏也影响了其翻译后调节因子——蛋白乙酰转移酶(patZ)。cobB(编码一种类沉默调节蛋白脱乙酰酶)和patZ基因缺失的影响取决于遗传背景。两种菌株中cobB的缺失均增加了乙酸盐的产生,并降低了乙酸盐培养基中的生长速率。BL21中patZ的缺失抑制了葡萄糖培养基中乙酸盐的溢流,并提高了乙酸盐培养基中的生长速率。BL21和K12菌株之间乙酸盐溢流的差异是由许多重叠因素引起的。确定了两个主要影响因素:(1)由于伴随产生的环磷酸腺苷,指数生长期acs在BL21菌株中不被抑制;(2)与K12相比,BL21中乙酰辅酶A合成酶的活性受蛋白乙酰化的调控更为严格。总之,这些差异导致了较低的乙酸盐溢流以及大肠杆菌BL21在葡萄糖存在时消耗这种代谢物的能力提高。

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