Two active forms of UDP-N-acetylglucosamine enolpyruvyl transferase in gram-positive bacteria.

Abstract:

:Gene sequences encoding the enzymes UDP-N-acetylglucosamine enolpyruvyl transferase (MurA) from many bacterial sources were analyzed. It was shown that whereas gram-negative bacteria have only one murA gene, gram-positive bacteria have two distinct genes encoding these enzymes which have possibly arisen from gene duplication. The two murA genes of the gram-positive organism Streptococcus pneumoniae were studied further. Each of the murA genes was individually inactivated by allelic replacement. In each case, the organism was viable despite losing one of its murA genes. However, when attempts were made to construct a double-deletion strain, no mutants were obtained. This indicates that both genes encode active enzymes that can substitute for each other, but that the presence of a MurA function is essential to the organism. The two genes were further cloned and overexpressed, and the enzymes they encode were purified. Both enzymes catalyzed the transfer of enolpyruvate from phosphoenolpyruvate to UDP-N-acetylglucosamine, confirming they are both active UDP-N-acetylglucosamine enolpyruvyl transferases. The catalytic parameters of the two enzymes were similar, and they were both inhibited by the antibiotic fosfomycin.

journal_name

J Bacteriol

journal_title

Journal of bacteriology

authors

Du W,Brown JR,Sylvester DR,Huang J,Chalker AF,So CY,Holmes DJ,Payne DJ,Wallis NG

doi

10.1128/jb.182.15.4146-4152.2000

keywords:

subject

Has Abstract

pub_date

2000-08-01 00:00:00

pages

4146-52

issue

15

eissn

0021-9193

issn

1098-5530

journal_volume

182

pub_type

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