2020 ; Vol.30-2: 271~278
|Author||Gyeong Soo Ko, Quyet Thang Nguyen, Do Hyeon Kim, Jin Kuk Yang|
|Place of duty||Department of Chemistry, College of Natural Sciences, Soongsil University, Seoul 06978, Republic of Korea|
|Title||Biochemical and Molecular Characterization of Glycerol Dehydrogenas from Klebsiella pneumoniae|
J. Microbiol. Biotechnol.2020 ;
|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 83.4oC, 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.|
|Key_word||Glycerol dehydrogenase, gldA, dihydroxyacetone production|
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