Elleuche, SkanderSkanderElleucheKlippel, BarbaraBarbaraKlippelHeyde, Amélie von derAmélie von derHeydeAntranikian, GarabedGarabedAntranikian2020-07-162020-07-162013-01-30Biotechnology Letters 5 (35): 725-733 (2013)http://hdl.handle.net/11420/6861Purpose of work: 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. © 2013 Springer Science+Business Media Dordrecht.en1573-677620135725733Springer Science + Business Media B.VButanalDickeya zeaeEthanolGene duplicationGroup III-alcohol dehydrogenaseSubstrate specificityBiowissenschaften, BiologieJournal Article10.1007/s10529-013-1137-2Other