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表达血影蛋白的重组大肠杆菌在微氧条件下优先进行需氧代谢:一项蛋白质组水平的研究。

Recombinant E. coli expressing Vitreoscilla haemoglobin prefers aerobic metabolism under microaerobic conditions: a proteome-level study.

机构信息

Department of Genetic Engineering, School of Biotechnology, Madurai Kamaraj University, Madurai 625 021.

出版信息

J Biosci. 2012 Sep;37(4):617-33. doi: 10.1007/s12038-012-9245-z.

Abstract

Vitreoscilla haemoglobin (VHb) expression in heterologous host was shown to enhance growth and oxygen utilization capabilities under oxygen-limited conditions. The exact mechanism by which VHb enhances the oxygen utilization under oxygen-limiting conditions is still unknown. In order to understand the role of VHb in promoting oxygen utilization, changes in the total protein profile of E. coli expressing the vgb gene under its native promoter was analysed. Two-dimensional difference gel electrophoresis (2D DIGE) was employed to quantify the differentially expressed proteins under oxygen-limiting conditions. Overexpression of proteins involved in aerobic metabolic pathways and suppression of proteins involved in non-oxidative metabolic pathways shown in this study indicates that the cells expressing VHb prefer aerobic metabolic pathways even under oxygen limitation. Under these conditions, the expression levels of proteins involved in central metabolic pathways, cellular adaptation and cell division were also found to be altered. These results imply that Vitreoscilla haemoglobin expression alters aerobic metabolism specifically, in addition to altering proteins involved in other pathways, the significance of which is not clear as of now.

摘要

在异源宿主中表达的血影蛋白(VHb)被证明可以在缺氧条件下增强生长和氧气利用能力。VHb 在缺氧条件下增强氧气利用的确切机制尚不清楚。为了了解 VHb 在促进氧气利用中的作用,分析了在其天然启动子下表达 vgb 基因的大肠杆菌的总蛋白图谱的变化。二维差异凝胶电泳(2D DIGE)用于定量缺氧条件下差异表达的蛋白质。本研究表明,有氧代谢途径中涉及的蛋白质表达增加,非氧化代谢途径中涉及的蛋白质表达受到抑制,这表明表达 VHb 的细胞即使在缺氧条件下也优先选择有氧代谢途径。在这些条件下,还发现参与中心代谢途径、细胞适应和细胞分裂的蛋白质的表达水平也发生了改变。这些结果表明,Vitreoscilla 血红蛋白的表达不仅改变了其他途径中涉及的蛋白质,还特异性地改变了有氧代谢,其意义目前尚不清楚。

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