Artie McFerrin Department of Chemical Engineering, Texas A & M University, College Station, TX 77843-3122, USA.
J Appl Microbiol. 2010 Jun;108(6):2088-102. doi: 10.1111/j.1365-2672.2009.04611.x. Epub 2009 Oct 30.
To complement our proteome study, whole-transcriptome analyses were utilized here to identify proteins related to degrading cis-1,2-dichloroethylene (cis-DCE).
Metabolically engineered Escherichia coli strains were utilized expressing an evolved toluene ortho-monooxygenase along with either (i) glutathione S-transferase and altered gamma-glutamylcysteine synthetase or (ii) a rationally engineered epoxide hydrolase. cis-DCE degradation induced 30 known stress genes and 32 uncharacterized genes. Because of the reactive cis-DCE epoxides formed, we hypothesized that some of these uncharacterized genes may be related to a variety of stresses. Using isogenic mutants, IbpB, YchH, YdeI, YeaR, YgiW, YoaG and YodD were related to hydrogen peroxide, cadmium and acid stress. Additional whole-transcriptome studies with hydrogen peroxide stress using the most hydrogen peroxide-sensitive mutants, ygiW and ychH, identified that FliS, GalS, HcaR, MglA, SufE, SufS, Tap, TnaB, YhcN and YjaA are also involved in the stress response of E. coli to hydrogen peroxide, cadmium and acid, as well as are involved in biofilm formation.
Seventeen proteins are involved in the stress network for this organism, and YhcN and YchH were shown to be important for the degradation of cis-DCE.
Six previously uncharacterized proteins (YchH, YdeI, YgiW, YhcN, YjaA and YodD) were shown to be stress proteins.
为了补充我们的蛋白质组学研究，本研究利用全转录组分析来鉴定与降解顺-1,2-二氯乙烯（cis-DCE）相关的蛋白质。
利用表达进化的甲苯邻位单加氧酶的代谢工程大肠杆菌菌株，同时表达（i）谷胱甘肽 S-转移酶和改变的γ-谷氨酰半胱氨酸合成酶，或（ii）合理设计的环氧化物水解酶。cis-DCE 降解诱导了 30 个已知的应激基因和 32 个未鉴定的基因。由于形成了反应性的 cis-DCE 环氧化物，我们假设其中一些未鉴定的基因可能与多种应激有关。使用同基因突变体，IbpB、YchH、YdeI、YeaR、YgiW、YoaG 和 YodD 与过氧化氢、镉和酸应激有关。使用最易受过氧化氢影响的突变体 ygiW 和 ychH 进行的额外全转录组研究表明，FliS、GalS、HcaR、MglA、SufE、SufS、Tap、TnaB、YhcN 和 YjaA 也参与了大肠杆菌对过氧化氢、镉和酸的应激反应，以及生物膜形成。
该生物体的应激网络涉及 17 种蛋白质，并且 YhcN 和 YchH 被证明对 cis-DCE 的降解很重要。
六个以前未鉴定的蛋白质（YchH、YdeI、YgiW、YhcN、YjaA 和 YodD）被证明是应激蛋白。
