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Characterization of the alanine racemases from two Mycobacteria

Characterization of the alanine racemases from two Mycobacteria Abstractd-Alanine is a necessary precursor in the biosynthesis of the bacterial peptidoglycan. The naturally occurring l-alanine isomer is racemized to its d-form through the action of a class of enzymes called alanine racemases. These enzymes are ubiquitous among prokaryotes, and with very few exceptions are absent in eukaryotes, making them a logical target for the development of novel antibiotics. The alanine racemase gene from both Mycobacterium tuberculosis and M. avium was amplified by PCR and cloned in Escherichia coli. Overexpression of the proteins in the E. coli BL21 system, both as native and as His-tagged recombinant products, has been achieved. The proteins have been purified to electrophoretic homogeneity and analyzed biochemically. A d-alanine requiring double knock-out mutant of E. coli (alr, dadX) was constructed and the cloned genes were able to complement its deficiencies. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png FEMS Microbiology Letters Oxford University Press

Characterization of the alanine racemases from two Mycobacteria

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References (23)

Publisher
Oxford University Press
Copyright
© 2001 Federation of European Microbiological Societies
ISSN
0378-1097
eISSN
1574-6968
DOI
10.1111/j.1574-6968.2001.tb10547.x
pmid
11267762
Publisher site
See Article on Publisher Site

Abstract

Abstractd-Alanine is a necessary precursor in the biosynthesis of the bacterial peptidoglycan. The naturally occurring l-alanine isomer is racemized to its d-form through the action of a class of enzymes called alanine racemases. These enzymes are ubiquitous among prokaryotes, and with very few exceptions are absent in eukaryotes, making them a logical target for the development of novel antibiotics. The alanine racemase gene from both Mycobacterium tuberculosis and M. avium was amplified by PCR and cloned in Escherichia coli. Overexpression of the proteins in the E. coli BL21 system, both as native and as His-tagged recombinant products, has been achieved. The proteins have been purified to electrophoretic homogeneity and analyzed biochemically. A d-alanine requiring double knock-out mutant of E. coli (alr, dadX) was constructed and the cloned genes were able to complement its deficiencies.

Journal

FEMS Microbiology Letters – Oxford University Press

Published: Mar 9, 2001

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