Overexpression, purification and characterization of Mycobacterium bovis BCG alcohol dehydrogenase.

Alcohol Dehydrogenase/genetics/isolation & purification/metabolism; Base Sequence; DNA Primers; Kinetics; Mycobacterium bovis/enzymology; Phenotype; Phylogeny

Abstract :

[en] A previous study of the effect of zinc deprivation on Mycobacterium bovis BCG pointed out the potential importance of an alcohol dehydrogenase for maintaining the hydrophobic character of the cell envelope. In this report, the effect of the overexpression of the M. bovis BCG alcohol dehydrogenase (ADH) in Mycobacterium smegmatis and M. bovis BCG is described. The purification of the enzyme was performed to apparent homogeneity from overexpressing M. bovis BCG cells and its kinetic parameters were determined. The enzyme showed a strong preference for both aromatic and aliphatic aldehydes while the corresponding alcohols were processed 100-1000-fold less efficiently. The best kcat/Km values were found with benzaldehyde > 3-methoxybenzaldehyde > octanal > coniferaldehyde. A phylogenetic analysis clearly revealed that the M. bovis BCG ADH together with the ADHs from Bacillus subtilis and Helicobacter pylori formed a sister group of the class C medium-chain alcohol dehydrogenases, the plant cinnamyl alcohol dehydrogenases (CADs). Comparison of the kinetic properties of our ADH with some related class C enzymes indicated that the mycobacterial enzyme substrate profile resembled that of the CADs involved in plant defence rather than those implicated in lignification. A possible role for the M. bovis BCG ADH in the biosynthesis of the lipids composing the mycobacterial cell envelope is proposed.

Disciplines :

Biochemistry, biophysics & molecular biology

Author, co-author :

Wilkin, J. M.

Soetaert, K.

Stelandre, M.

Buyssens, P.

Castillo Cabello, Gabriel ; Université de Liège - ULiège > Département des sciences de la vie > Département des sciences de la vie

Demoulin, Vincent

Bottu, G.

Laneelle, M. A.

Daffe, M.

De Bruyn, J.

Language :

English

Title :

Overexpression, purification and characterization of Mycobacterium bovis BCG alcohol dehydrogenase.

Publication date :

1999

Journal title :

European Journal of Biochemistry

ISSN :

0014-2956

eISSN :

1432-1033

Publisher :

Blackwell Science, Oxford, United Kingdom

Volume :

262

Issue :

Pages :

299-307

Peer reviewed :

Peer Reviewed verified by ORBi

Available on ORBi :

since 09 November 2011

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