Cloning and characterization of a ribitol dehydrogenase from Zymomonas mobilis

Applied Microbiology and Biotechnology
Hee-Jung MoonJung-Kul Lee

Abstract

Ribitol dehydrogenase (RDH) catalyzes the conversion of ribitol to D-ribulose. A novel RDH gene was cloned from Zymomonas mobilis subsp. mobilis ZM4 and overexpressed in Escherichia coli BL21(DE3). DNA sequence analysis revealed an open reading frame of 795 bp, capable of encoding a polypeptide of 266 amino acid residues with a calculated molecular mass of 28,426 Da. The gene was overexpressed in E. coli BL21(DE3) and the protein was purified as an active soluble form using glutathione S-transferase affinity chromatography. The molecular mass of the purified enzyme was estimated to be approximately 28 kDa by sodium dodecyl sulfate-polyacrylamide gel and approximately 58 KDa with gel filtration chromatography, suggesting that the enzyme is a homodimer. The enzyme had an optimal pH and temperature of 9.5 and 65 degrees C, respectively. Unlike previously characterized RDHs, Z. mobilis RDH (ZmRDH) showed an unusual dual coenzyme specificity, with a k(cat) of 4.83 s(-1) for NADH (k(cat)/K(m) = 27.3 s(-1) mM(-1)) and k(cat) of 2.79 s(-1) for NADPH (k(cat)/K(m) = 10.8 s(-1) mM(-1)). Homology modeling and docking studies of NAD+ and NADP+ into the active site of ZmRDH shed light on the dual coenzyme specificity of ZmRDH.

References

Sep 1, 1974·The Biochemical Journal·S S TaylorB S Hartley
May 9, 1995·Biochemistry·H JörnvallD Ghosh
Jun 7, 1994·Proceedings of the National Academy of Sciences of the United States of America·K I VarugheseJ M Whiteley
Jan 1, 1997·Advances in Experimental Medicine and Biology·U C OppermannH Jörnvall
Jan 1, 1999·Bioorganic & Medicinal Chemistry Letters·L S Jeong, S J Yoo
Aug 17, 1999·Journal of Protein Chemistry·M I Homsi-BrandeburgoB S Hartley
Jan 27, 2000·World Review of Nutrition and Dietetics·H BertelsenB Buemann
Dec 1, 2001·Biochemical and Biophysical Research Communications·C FillingU Oppermann
Apr 27, 2002·The Journal of Biological Chemistry·Charlotta FillingUdo Oppermann
Feb 27, 2003·Chemico-biological Interactions·Udo OppermannHans Jörnvall
Dec 14, 2004·Nature Biotechnology·Jeong-Sun SeoHyen Sam Kang
Mar 15, 2006·Structure·Andreas H EhrensbergerDavid K Wilson

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