Comparative analysis of two members of the metal ion-containing group III-alcohol dehydrogenases from Dickeya zeae

Journal
Biotechnology letters  
Volume
35
Issue
5
Start Page
725
End Page
733
Citation
Biotechnology Letters 5 (35): 725-733 (2013)
Publisher DOI
10.1007/s10529-013-1137-2
Scopus ID
2-s2.0-84876976947
Publisher
Springer Science + Business Media B.V
Purpose 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.
Subjects
Butanal
Dickeya zeae
Ethanol
Gene duplication
Group III-alcohol dehydrogenase
Substrate specificity
DDC Class
570: Biowissenschaften, Biologie
Funding(s)
Fundamentals for synthetic biological systems  
More Funding Information
Excellence Cluster in the Excellence Initative by the State of Hamburg