An efficient ribitol-specific dehydrogenase from Enterobacter aerogenes

Ranjitha Singh, Raushan Singh, In Won Kim, Sujan Sigdel, Vipin C. Kalia, Yun Chan Kang, Jung Kul Lee

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

An NAD+-dependent ribitol dehydrogenase from Enterobacter aerogenes KCTC 2190 (EaRDH) was cloned and successfully expressed in Escherichia coli. The complete 729-bp gene was amplified, cloned, expressed, and subsequently purified in an active soluble form using nickel affinity chromatography. The enzyme had an optimal pH and temperature of 11.0 and 45°C, respectively. Among various polyols, EaRDH exhibited activity only toward ribitol, with Km, Vmax, and kcat/Km values of 10.3mM, 185Umg-1, and 30.9s-1mM-1, respectively. The enzyme showed strong preference for NAD+ and displayed no detectable activity with NADP+. Homology modeling and sequence analysis of EaRDH, along with its biochemical properties, confirmed that EaRDH belongs to the family of NAD+-dependent ribitol dehydrogenases, a member of short-chain dehydrogenase/reductase (SCOR) family. EaRDH showed the highest activity and unique substrate specificity among all known RDHs. Homology modeling and docking analysis shed light on the molecular basis of its unusually high activity and substrate specificity.

Original languageEnglish
Pages (from-to)56-64
Number of pages9
JournalEnzyme and Microbial Technology
Volume72
DOIs
Publication statusPublished - 2015 May 1

Fingerprint

ribitol 2-dehydrogenase
Ribitol
Enterobacter aerogenes
NAD
Oxidoreductases
Substrate Specificity
Affinity chromatography
Enzymes
Substrates
Nickel
NADP
Affinity Chromatography
Escherichia coli
Sequence Analysis
Polyols
Genes
Temperature

Keywords

  • Enterobacter aerogenes
  • Homology modeling
  • Ribitol dehydrogenase
  • Short-chain dehydrogenase/reductase

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Biochemistry
  • Applied Microbiology and Biotechnology

Cite this

An efficient ribitol-specific dehydrogenase from Enterobacter aerogenes. / Singh, Ranjitha; Singh, Raushan; Kim, In Won; Sigdel, Sujan; Kalia, Vipin C.; Kang, Yun Chan; Lee, Jung Kul.

In: Enzyme and Microbial Technology, Vol. 72, 01.05.2015, p. 56-64.

Research output: Contribution to journalArticle

Singh, Ranjitha ; Singh, Raushan ; Kim, In Won ; Sigdel, Sujan ; Kalia, Vipin C. ; Kang, Yun Chan ; Lee, Jung Kul. / An efficient ribitol-specific dehydrogenase from Enterobacter aerogenes. In: Enzyme and Microbial Technology. 2015 ; Vol. 72. pp. 56-64.
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