Molecular cloning, sequence analysis and functional characterization of the gene kdsA, encoding 3-deoxy-D-manno-2-octulosonate-8-phosphate synthase of Chlamydia psittaci 6BC

被引：12

作者：

Brabetz, W

Brade, H

机构：

[1] Division of Biochemical Microbiology, Research Center Borstel, Center for Medicine and Biosciences

[2] Division of Biochemical Microbiology, Research Center Borstel, D-23845 Borstel

来源：

EUROPEAN JOURNAL OF BIOCHEMISTRY | 1997年 / 244卷 / 01期

关键词：

3-deoxy-D-manno-2-octulosonate-8-phosphate synthase; lipopolysaccharide biosynthesis; Chlamydia;

D O I：

10.1111/j.1432-1033.1997.00066.x

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

The kdsA gene encoding 3-deoxy-D-manno-2-octulosenate-8-phosphate (Kdo-8-P) synthase of Chlamydia psittaci 6BC was cloned by complementing the temperature-sensitive kdsA mutant Salmonella enterica serovar Typhimurium AG701i50. The sequence analysis of a recombinant DNA fragment revealed an open reading frame of 807 nucleotides which codes for a polypeptide of 269 amino acids with a high degree of similarity to known KdsA proteins. In addition, alignments of Kdo-8-P synthases with bacterial and fungal 3-deoxy-D-arabino-2-heptulosonate-7-phosphate (Dha-7-P) synthases suggested that both classes of enzymes are structurally related and may belong to a family of 2-keto-3-deoxy-aldonic acid synthases. The chlamydial protein was overexpressed and functionally characterized in vitro to synthesize Kdo-8-P from D-arabinose 5-phosphate and phosphoenolpyruvate. A chlamydial DNA region upstream of the gene exhibiting similarities to the consensus sequence of sigma(70) promoters of Escherichia coli was responsible for the heterologous expression of kdsA.

引用

页码：66 / 73

页数：8

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