(R,R)-Butane-2,3-diol Dehydrogenase from Bacillus clausii DSM 8716T: Cloning and Expression of the bdhA-Gene, and Initial Characterization of Enzyme

  • The gene encoding a putative (R,R)-butane-2,3-diol dehydrogenase (bdhA) from Bacillus clausii DSM 8716T was isolated, sequenced and expressed in Escherichia coli. The amino acid sequence of the encoded protein is only distantly related to previously studied enzymes (identity 33–43%) and exhibited some uncharted peculiarities. An N-terminally StrepII-tagged enzyme variant was purified and initially characterized. The isolated enzyme catalyzed the (R)-specific oxidation of (R,R)- and meso-butane-2,3-diol to (R)- and (S)-acetoin with specific activities of 12 U/mg and 23 U/mg, respectively. Likewise, racemic acetoin was reduced with a specific activity of up to 115 U/mg yielding a mixture of (R,R)- and meso-butane-2,3-diol, while the enzyme reduced butane-2,3-dione (Vmax 74 U/mg) solely to (R,R)-butane-2,3-diol via (R)-acetoin. For these reactions only activity with the co-substrates NADH/NAD+ was observed. The enzyme accepted a selection of vicinal diketones, α-hydroxy ketones and vicinal diols as alternative substrates. Although the physiological function of the enzyme in B. clausii remains elusive, the data presented herein clearly demonstrates that the encoded enzyme is a genuine (R,R)-butane-2,3-diol dehydrogenase with potential for applications in biocatalysis and sensor development.

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Author:Lukas Muschallik, Denise MolinnusORCiD, Johannes BongaertsORCiD, Martina Pohl, Torsten WagnerORCiD, Michael Josef SchöningORCiD, Petra SiegertORCiD, Thorsten Selmer
DOI:https://doi.org/10.1016/j.jbiotec.2017.07.020
ISSN:0168-1656
Parent Title (English):Journal of Biotechnology
Publisher:Elsevier
Place of publication:Amsterdam
Document Type:Article
Language:English
Year of Completion:2017
Volume:258
First Page:41
Last Page:50
Link:https://doi.org/10.1016/j.jbiotec.2017.07.020
Zugriffsart:campus
Institutes:FH Aachen / Fachbereich Chemie und Biotechnologie
FH Aachen / Fachbereich Medizintechnik und Technomathematik
FH Aachen / INB - Institut für Nano- und Biotechnologien
collections:Verlag / Elsevier