Construction and characterization of hybrid dehydratases between adenosylcobalamin-dependent diol and glycerol dehydratases

Takahiro Sakai, Ai Yamasaki, Satoshi Toyofuku, Tsuneo Nishiki, Michio Yunoki, Noriaki Komoto, Takamasa Tobimatsu, Tetsuo Toraya

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Adenosylcobalamin-dependent diol dehydratase and glycerol dehydratase are isofunctional enzymes that catalyze the dehydration of 1,2-diols to the corresponding aldehydes. Although they bear different metabolic roles, both enzymes consist of three different subunits and possess a common (αβγ)2 structure. To elucidate the roles of each subunit, we constructed expression plasmids for the hybrid dehydratases between diol dehydratase of Klebsiella oxytoca and glycerol dehydratase of Klebsiella pneumoniae in all the combinations of subunits by gene engineering techniques. All of the hybrid enzymes were produced in Escherichia coli at high levels, but only two hybrid enzymes consisting of the α subunit from glycerol dehydratase and the β subunits from diol dehydratase showed high activity. The substrate specificity, the susceptibility to inactivation by glycerol, and the monovalent cation specificity of the wild type and hybrid enzymes were primarily determined by the origin of their α subunits.

Original languageEnglish
Pages (from-to)102-108
Number of pages7
JournalJournal of Nutritional Science and Vitaminology
Volume53
Issue number2
DOIs
Publication statusPublished - Apr 2007

Fingerprint

glycerol dehydratase
Propanediol Dehydratase
Hydro-Lyases
glycols
glycerol
Enzymes
enzymes
Klebsiella oxytoca
Klebsiella pneumoniae
protein subunits
substrate specificity
Monovalent Cations
aldehydes
cations
plasmids
inactivation
Substrate Specificity
engineering
Dehydration
Aldehydes

Keywords

  • Adenosylcobalamin
  • Coenzyme B
  • Diol dehydratase
  • Glycerol dehydratase
  • Hybrid enzymes

ASJC Scopus subject areas

  • Medicine (miscellaneous)
  • Food Science

Cite this

Construction and characterization of hybrid dehydratases between adenosylcobalamin-dependent diol and glycerol dehydratases. / Sakai, Takahiro; Yamasaki, Ai; Toyofuku, Satoshi; Nishiki, Tsuneo; Yunoki, Michio; Komoto, Noriaki; Tobimatsu, Takamasa; Toraya, Tetsuo.

In: Journal of Nutritional Science and Vitaminology, Vol. 53, No. 2, 04.2007, p. 102-108.

Research output: Contribution to journalArticle

Sakai, Takahiro ; Yamasaki, Ai ; Toyofuku, Satoshi ; Nishiki, Tsuneo ; Yunoki, Michio ; Komoto, Noriaki ; Tobimatsu, Takamasa ; Toraya, Tetsuo. / Construction and characterization of hybrid dehydratases between adenosylcobalamin-dependent diol and glycerol dehydratases. In: Journal of Nutritional Science and Vitaminology. 2007 ; Vol. 53, No. 2. pp. 102-108.
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AU - Yunoki, Michio

AU - Komoto, Noriaki

AU - Tobimatsu, Takamasa

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