期刊
JOURNAL OF BACTERIOLOGY
卷 187, 期 15, 页码 5061-5066出版社
AMER SOC MICROBIOLOGY
DOI: 10.1128/JB.187.15.5061-5066.2005
关键词
-
类别
资金
- Biotechnology and Biological Sciences Research Council [BB/C513918/1] Funding Source: researchfish
- Biotechnology and Biological Sciences Research Council [BB/C513918/1] Funding Source: Medline
In some bacteria, salicylate is synthesized using the enzymes isochorismate synthase and isochorismate pyruvate lyase. In contrast, gene inactivation and complementation experiments with Yersinia enterocolitica suggest the synthesis of salicylate in the biosynthesis of the siderophore yersiniabactin involves a single protein, Irp9, which converts chorismate directly into salicylate. In the present study, Irp9 was for the first time heterologously expressed in Escherichia coli as a hexahistidine fusion protein, purified to near homogeneity, and characterized biochemically. The recombinant protein was found to be a dimer, each subunit of which has a molecular mass of 50 kDa. Enzyme assays, reverse-phase high-pressure liquid chromatography and H-1 nuclear magnetic resonance (NMR) spectroscopic analyses confirmed that Irp9 is a salicylate synthase and converts chorismate to salicylate with a K for chorismate of 4.2 mu M and a k(cat) of 8 min(-1). The reaction was shown to proceed through the intermediate isochorismate, which was detected directly using H-1 NMR spectroscopy.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据