Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Biochemical and Molecular Characterization of Glycerol Dehydrogenase from Klebsiella pneumoniae

  • Ko, Gyeong Soo (Department of Chemistry, College of Natural Sciences, Soongsil University) ;
  • Nguyen, Quyet Thang (Department of Chemistry, College of Natural Sciences, Soongsil University) ;
  • Kim, Do Hyeon (Department of Chemistry, College of Natural Sciences, Soongsil University) ;
  • Yang, Jin Kuk (Department of Chemistry, College of Natural Sciences, Soongsil University)
  • Received : 2019.09.30
  • Accepted : 2019.10.21
  • Published : 2020.02.28
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Abstract

Glycerol dehydrogenase (GlyDH) catalyzes the oxidation of glycerol to dihydroxyacetone (DHA), which is the first step in the glycerol metabolism pathway. GlyDH has attracted great interest for its potential industrial applications, since DHA is a precursor for the synthesis of many commercially valuable chemicals and various drugs. In this study, GlyDH from Klebsiella pneumoniae (KpGlyDH) was overexpressed in E. coli and purified to homogeneity for biochemical and molecular characterization. KpGlyDH exhibits an exclusive preference for NAD+ over NADP+. The enzymatic activity of KpGlyDH is maximal at pH 8.6 and pH 10.0. Of the three common polyol substrates, KpGlyDH showed the highest kcat/Km value for glycerol, which is three times higher than for racemic 2,3-butanediol and 32 times higher than for ethylene glycol. The kcat value for glycerol oxidation is notably high at 87.1 ± 11.3 sec-1. KpGlyDH was shown to exist in an equilibrium between two different oligomeric states, octamer and hexadecamer, by size-exclusion chromatography analysis. KpGlyDH is structurally thermostable, with a Tm of 83.4℃, in thermal denaturation experiment using circular dichroism spectroscopy. The biochemical and biophysical characteristics of KpGlyDH revealed in this study should provide the basis for future research on its glycerol metabolism and possible use in industrial applications.

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