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Adaptive laboratory evolution of Escherichia coli lacking cellular byproduct formation for enhanced acetate utilization through compensatory ATP consumption.
Metabolic Engineering ( IF 6.8 ) Pub Date : 2020-09-12 , DOI: 10.1016/j.ymben.2020.09.005
Wonjae Seong 1 , Gui Hwan Han 2 , Hyun Seung Lim 3 , Ji In Baek 4 , Soo-Jung Kim 5 , Donghyuk Kim 6 , Seong Keun Kim 3 , Hyewon Lee 3 , Haseong Kim 1 , Seung-Goo Lee 1 , Dae-Hee Lee 1
Affiliation  

Acetate has attracted great attention as a carbon source to develop economically feasible bioprocesses for sustainable bioproducts. Acetate is a less-preferred carbon source and a well-known growth inhibitor of Escherichia coli. In this study, we carried out adaptive laboratory evolution of an E. coli strain lacking four genes (adhE, pta, ldhA, and frdA) involved in acetyl-CoA consumption, allowing the efficient utilization of acetate as its sole carbon and energy source. Four genomic mutations were found in the evolved strain through whole-genome sequencing, and two major mutations (in cspC and patZ) mainly contributed to efficient utilization of acetate and tolerance to acetate. Transcriptomic reprogramming was examined by analyzing the genome-wide transcriptome with different carbon sources. The evolved strain showed high levels of intracellular ATP by upregulation of genes involved in NADH and ATP biosynthesis, which facilitated the production of enhanced green fluorescent protein, mevalonate, and n-butanol using acetate alone. This new strain, given its high acetate tolerance and high ATP levels, has potential as a starting host for cell factories targeting the production of acetyl-CoA-derived products from acetate or of products requiring high ATP levels.



中文翻译:

缺乏细胞副产物形成的大肠杆菌的适应性实验室进化,通过补偿性 ATP 消耗提高乙酸盐利用率。

醋酸盐作为一种碳源已经引起了极大的关注,以开发可持续生物产品的经济可行的生物工艺。醋酸盐是不太优选的碳源,也是众所周知的大肠杆菌生长抑制剂。在这项研究中,我们对缺乏参与乙酰辅酶 A 消耗的四个基因(adhEptaldhAfrdA)的大肠杆菌菌株进行了适应性实验室进化,从而能够有效利用乙酸盐作为其唯一的碳源和能源。通过全基因组测序在进化菌株中发现了四个基因组突变,以及两个主要突变(在cspCpatZ) 主要有助于醋酸盐的有效利用和对醋酸盐的耐受性。通过分析具有不同碳源的全基因组转录组来检查转录组重编程。进化的菌株通过上调参与 NADH 和 ATP 生物合成的基因显示出高水平的细胞内 ATP,这促进了增强型绿色荧光蛋白、甲羟戊酸和正丁醇的产生。这种新菌株具有高乙酸盐耐受性和高 ATP 水平,有可能成为细胞工厂的起始宿主,这些细胞工厂的目标是从乙酸盐生产乙酰辅酶 A 衍生产品或需要高 ATP 水平的产品。

更新日期:2020-09-30
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