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Characterization of two aldo–keto reductases from Gluconobacter oxydans 621H capable of regio- and stereoselective α-ketocarbonyl reduction

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Abstract

Two cytosolic NADPH-dependent carbonyl reductases from Gluconobacter oxydans 621H, Gox0644 and Gox1615, were heterologously produced in Escherichia coli. The recombinant proteins were purified to homogeneity and characterized. Gox0644 and Gox1615 were dimers with native molecular masses of 66.1 and 74.5 kDa, respectively. The enzymes displayed broad substrate specificities and reduced α-ketocarbonyls at the keto moiety most proximal to the terminus of the alkyl chain to produce alpha-hydroxy carbonyls, as demonstrated by NMR. With respect to stereoselectivity, protein Gox0644 specifically reduced 2,3-pentanedione to 2R-hydroxy-pentane-3-one, whereas Gox1615 produced 2S-hydroxy-pentane-3-one. Both enzymes also reduced 1-phenyl-1,2-propanedione to 2-hydroxy-1-phenylpropane-1-one, which is a key intermediate in the production of numerous pharmaceuticals, such as antifungal azoles and antidepressants. Gox0644 displayed highest activities with 2,3-diones, α-ketoaldehydes, α-keto esters, and 2,5-diketogluconate. Gox1615 was less active with these substrates, but displayed a broader substrate spectrum reducing a variety of α-diketones and aldehydes.

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Acknowledgments

The authors would like to thank Jana Moldenrhauer, Kekulé Institute of Organic Chemistry and Biochemistry, University of Bonn for her valuable technical advice during the synthesis of the chiral NMR solvent shift reagent. Thanks also goes to Dr. Engeser and her team, Kekulé Institute of Organic Chemistry and Biochemistry, University of Bonn for performing MS measurements. This project as supported by funds from Bundesministerium für Bildung und Forschung (BMBF, project no. 0315632A).

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  1. Institute für Mikrobiologie und Biotechnologie, 168 Meckenheimer Allee, 53115, Bonn, Germany

    Paul Schweiger & Uwe Deppenmeier

  2. Institut für Pharmazeutische Biologie, Nußallee 6, 53115, Bonn, Germany

    Harald Gross

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Schweiger, P., Gross, H. & Deppenmeier, U. Characterization of two aldo–keto reductases from Gluconobacter oxydans 621H capable of regio- and stereoselective α-ketocarbonyl reduction. Appl Microbiol Biotechnol 87, 1415–1426 (2010). https://doi.org/10.1007/s00253-010-2607-2

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