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丙酮丁醇梭菌ATCC 824中分解代谢物阻遏元件的分析

Analysis of the elements of catabolite repression in Clostridium acetobutylicum ATCC 824.

作者信息

Tangney Martin, Galinier Anne, Deutscher Josef, Mitchell Wilfrid J

机构信息

School of Life Sciences, Napier University, Edinburgh, UK.

出版信息

J Mol Microbiol Biotechnol. 2003;6(1):6-11. doi: 10.1159/000073403.

Abstract

The PTSH gene, encoding the phosphotransferase protein HPr, from Clostridium acetobutylicum ATCC 824 was identified from the genome sequence, cloned and shown to complement a PTSH mutant of Escherichia coli. The deduced protein sequence shares significant homology with HPr proteins from other low-GC gram-positive bacteria, although the highly conserved sequence surrounding the Ser-46 phosphorylation site is not well preserved in the clostridial protein. Nevertheless, the HPr was phosphorylated in an ATP-dependent manner in cell-free extracts of C. Acetobutylicum. Furthermore, purified His-tagged HPr from Bacillus Subtilis was also a substrate for the clostridial HPr kinase/phosphorylase. This phosphorylation reaction is a key step in the mechanism of carbon catabolite repression proposed to operate in B. Subtilis and other low-GC gram-positive bacteria. Putative genes encoding the HPr kinase/phosphorylase and the other element of this model, namely the catabolite control protein CcpA, were identified from the C. Acetobutylicum genome sequence, suggesting that a similar mechanism of carbon catabolite repression may operate in this industrially important organism.

摘要

从丙酮丁醇梭菌ATCC 824的基因组序列中鉴定出编码磷酸转移酶蛋白HPr的PTSH基因,进行克隆并证明其可互补大肠杆菌的PTSH突变体。推导的蛋白质序列与其他低GC含量革兰氏阳性细菌的HPr蛋白具有显著同源性,尽管在梭菌蛋白中Ser-46磷酸化位点周围的高度保守序列保存不佳。然而,在丙酮丁醇梭菌的无细胞提取物中,HPr以ATP依赖的方式被磷酸化。此外,从枯草芽孢杆菌纯化的His标签HPr也是梭菌HPr激酶/磷酸酶的底物。这种磷酸化反应是在枯草芽孢杆菌和其他低GC含量革兰氏阳性细菌中提出的碳分解代谢物阻遏机制的关键步骤。从丙酮丁醇梭菌基因组序列中鉴定出编码HPr激酶/磷酸酶以及该模型的另一个元件即分解代谢物控制蛋白CcpA的推定基因,这表明在这种具有重要工业价值的生物体中可能存在类似的碳分解代谢物阻遏机制。

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