Molecular Characterization of an NADPH-Dependent Acetoin Reductase/2,3-Butanediol Dehydrogenase from Clostridium beijerinckii NCIMB 8052

Author:

Raedts John1,Siemerink Marco A. J.1,Levisson Mark1,van der Oost John1,Kengen Servé W. M.1

Affiliation:

1. Laboratory of Microbiology, Wageningen University and Research Centre, Wageningen, The Netherlands

Abstract

ABSTRACT Acetoin reductase is an important enzyme for the fermentative production of 2,3-butanediol, a chemical compound with a very broad industrial use. Here, we report on the discovery and characterization of an acetoin reductase from Clostridium beijerinckii NCIMB 8052. An in silico screen of the C. beijerinckii genome revealed eight potential acetoin reductases. One of them ( CBEI_1464 ) showed substantial acetoin reductase activity after expression in Escherichia coli . The purified enzyme ( C. beijerinckii acetoin reductase [Cb-ACR]) was found to exist predominantly as a homodimer. In addition to acetoin (or 2,3-butanediol), other secondary alcohols and corresponding ketones were converted as well, provided that another electronegative group was attached to the adjacent C-3 carbon. Optimal activity was at pH 6.5 (reduction) and 9.5 (oxidation) and around 68°C. Cb-ACR accepts both NADH and NADPH as electron donors; however, unlike closely related enzymes, NADPH is preferred ( K m , 32 μM). Cb-ACR was compared to characterized close homologs, all belonging to the “threonine dehydrogenase and related Zn-dependent dehydrogenases” (COG1063). Metal analysis confirmed the presence of 2 Zn 2+ atoms. To gain insight into the substrate and cofactor specificity, a structural model was constructed. The catalytic zinc atom is likely coordinated by Cys 37 , His 70 , and Glu 71 , while the structural zinc site is probably composed of Cys 100 , Cys 103 , Cys 106 , and Cys 114 . Residues determining NADP specificity were predicted as well. The physiological role of Cb-ACR in C. beijerinckii is discussed.

Publisher

American Society for Microbiology

Subject

Ecology,Applied Microbiology and Biotechnology,Food Science,Biotechnology

Reference59 articles.

1. Lopez ContrerasAM, KuitW, SiemerinkMAJ, KengenSWM, SpringerJ and ClaassenPAM. 2010. Production of longer-chain alcohols from biomass—butanol, isopropanol and 2,3-butanediol, p 415–460. In WaldronK (ed), Bioalcohol production. Woodhead Publishing Ltd, Cambridge, United Kingdom.

2. Acetone-butanol fermentation revisited

3. Agitation and pressure effects on acetone-butanol fermentation

4. Microbial 2,3-butanediol production: A state-of-the-art review

5. Acetoin Metabolism in Bacteria

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