DNA microarray of global transcription factor mutant reveals membrane-related proteins involved in n-butanol tolerance in Escherichia coli.
DNA microarray of global transcription factor mutant reveals membrane-related proteins involved in n-butanol tolerance in Escherichia coli.
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全转录因子突变体的DNA微阵列揭示了大肠杆菌中参与正丁醇耐受的膜相关蛋白
DOI：
10.1186/s13068-016-0527-9
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发表时间：
2016
影响因子：
6.3
通讯作者：
Ni Y
中科院分区：
文献类型：
--
作者：
Si HM;Zhang F;Wu AN;Han RZ;Xu GC;Ni Y
背景：大肠杆菌已被探索作为丁醇等生物燃料生产的平台宿主菌株。然而，丁醇的严重毒性被认为是大肠杆菌生产丁醇的主要限制之一。因此，本研究的目的是构建耐丁醇的大肠杆菌菌株并阐明其耐受机制。结果：通过全局转录机器工程（gTME）方法分离出一株具有σ（70）突变、能够耐受2%（v/v）丁醇的重组大肠杆菌菌株。 DNA 微阵列用于评估丁醇耐受菌株 B8 的转录组谱。与野生型菌株相比，鉴定出 329 个差异表达基因（197 个上调，132 个下调）（p < 0.05；FC ≥ 2）。这些基因涉及碳水化合物代谢、能量代谢、双组分信号转导系统、氧化应激反应、脂质和细胞膜生物发生和外排泵。结论：几种膜相关蛋白被证明参与大肠杆菌的丁醇耐受。两个下调基因 yibT 和 yghW 被确定能够通过调节膜脂肪酸组成来影响丁醇耐受性。另一个下调的基因 ybjC 编码预测的内膜蛋白。此外，一些上调的基因，如gcl和glcF，有助于补充乙醛酸和TCA循环的代谢中间体，以增强能量供应。我们的结果可以作为构建作为生物燃料生产者的大肠杆菌菌株平台的实用策略。
Background:Escherichia coli has been explored as a platform host strain for biofuels production such as butanol. However, the severe toxicity of butanol is considered to be one major limitation for butanol production from E. coli. The goal of this study is therefore to construct butanol-tolerant E. coli strains and clarify the tolerance mechanisms.Results:A recombinant E. coli strain harboring σ(70) mutation capable of tolerating 2 % (v/v) butanol was isolated by the global transcription machinery engineering (gTME) approach. DNA microarrays were employed to assess the transcriptome profile of butanol-tolerant strain B8. Compared with the wild-type strain, 329 differentially expressed genes (197 up-regulated and 132 down-regulated) (p < 0.05; FC ≥ 2) were identified. These genes are involved in carbohydrate metabolism, energy metabolism, two-component signal transduction system, oxidative stress response, lipid and cell envelope biogenesis and efflux pump.Conclusions:Several membrane-related proteins were proved to be involved in butanol tolerance of E. coli. Two down-regulated genes, yibT and yghW, were identified to be capable of affecting butanol tolerance by regulating membrane fatty acid composition. Another down-regulated gene ybjC encodes a predicted inner membrane protein. In addition, a number of up-regulated genes, such as gcl and glcF, contribute to supplement metabolic intermediates for glyoxylate and TCA cycles to enhance energy supply. Our results could serve as a practical strategy for the construction of platform E. coli strains as biofuel producer.