Comparative analysis of two members of the metal ion-containing group III-alcohol dehydrogenases from Dickeya zeae.
A pair of NAD(+)- and NADP(+)-dependent group III-alcohol dehydrogenases was characterized from the enterobacterium, Dickeya zeae, to expand our understanding of the distribution and biochemical properties of this interesting group of enzymes. Two putative group III-alcohol dehydrogenases (ADHs) were identified in the genome of Dickeya zeae. Amino acid alignments and phylogenetic analysis revealed that Adh3.1 and Adh3.2 are only distantly related (~25 % identity at the protein level). Both proteins were purified to homogeneity after heterologous expression in E. coli. A specific activity of 1.8 U/mg was measured for the NAD(+)-dependent enzyme Adh3.1 with ethanol used as substrate, while NADPH-dependent Adh3.2 preferred butanal (29.1 U/mg) as substrate. Maximum activity for Adh3.1 was at 50 °C and pH 10 and for Adh3.2 at 70 °C and pH 6. Cell viability assays were used to confirm activity towards butanal and glyoxals. Biochemical characterization and phylogenetic analyses led to the hypothesis that Adh3.1 and Adh3.2 are probably the result of an ancient gene duplication event followed by functional diversification.
Institute of Technical Microbiology, Hamburg University of Technology (TUHH), Kasernenstr. 12, 21073 Hamburg, Germany. skander.elleuche@tuhh.de
Skander
S
Elleuche
Barbara
B
Klippel
Amélie
A
von der Heyde
Garabed
G
Antranikian
D000445:Aldehyde Oxidoreductases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D000447:Aldehydes / Q000378:metabolism / Q000494:pharmacology; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D004755:Enterobacteriaceae / Q000201:enzymology* / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D000431:Ethanol / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D050296:Microbial Viability / Q000187:drug effects; D010802:Phylogeny; D011765:Pyruvaldehyde / Q000378:metabolism / Q000494:pharmacology; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013379:Substrate Specificity; D013696:Temperature
D000447:Aldehydes; D001426:Bacterial Proteins; D011994:Recombinant Proteins; D000431:Ethanol; D011765:Pyruvaldehyde; C020145:formaldehyde dehydrogenase (glutathione); D000445:Aldehyde Oxidoreductases; C018475:butyraldehyde