PMID: 2493267Feb 23, 1989Paper

Isolation and characterisation of the glycerol dehydrogenase from Bacillus stearothermophilus

Biochimica Et Biophysica Acta
P SpencerM G Gore

Abstract

A protocol for the rapid purification of the glycerol dehydrogenase (glycerol: NAD+ 2-oxidoreductase, EC 1.1.1.6) from the thermophile Bacillus stearothermophilus has been developed using a combination of chromatographic techniques including affinity chromatography on a Sepharose-immobilised triazine dye (Procion red, HE3B, ICI). Substrate specificity has been examined and Km values determined. The protein has been shown to have an oligomeric Mr of approx. 180,000 and consists of four identical subunits of Mr 42,000. Exposure to chelating agents (e.g., EDTA) leads to total loss of activity; the EDTA-inactivated enzyme can be reactivated by Zn2+ and requires 1 mol equivalent of zinc per subunit for full catalytic activity. Other divalent cations such as Cd2+ and Co2+ will reactivate the apo-enzyme but yields an enzyme of lower specific activity. The enzyme binds 1 equivalent of NADH per subunit and during catalysis transfers the 4-pro-R hydride from the nicotinamide ring of the reduced-coenzyme to the substrate. Glycerol increases the dissociation constant for the interaction between NADH and Zn-metallo-glycerol dehydrogenase (ZnGDH) but has no effect on the equilibrium between NADH and metal-depleted enzyme.

References

Feb 17, 1975·Biochemical and Biophysical Research Communications·M G Gore, C Greenwood
Mar 1, 1978·Chemical & Pharmaceutical Bulletin·M SugiuraF Hirata
Apr 1, 1973·The Biochemical Journal·R A Stinson, J J Holbrook
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Citations

Apr 17, 2010·Applied Microbiology and Biotechnology·Tomohiko MatsuzawaKaoru Takegawa
Jan 8, 1999·Biochimica Et Biophysica Acta·N S PrestonM G Gore
Nov 9, 2011·Proceedings of the National Academy of Sciences of the United States of America·Qingzhao WangK T Shanmugam
Apr 3, 2004·Proceedings. Biological Sciences·M Claire Horner-DevineBrendan J M Bohannan
May 7, 2005·Archaea : an International Microbiological Journal·Jin-Suk Han, Kazuhiko Ishikawa
Mar 19, 2014·Acta Crystallographica. Section F, Structural Biology Communications·Paul Musille, Eric Ortlund
Mar 7, 2019·Acta Crystallographica. Section F, Structural Biology Communications·Jun ZhangYan Zhang
Jan 1, 1994·Critical Reviews in Microbiology·M F Reid, C A Fewson
Mar 26, 2014·Applied Microbiology and Biotechnology·Justin BeauchampClaire Vieille
Oct 9, 2002·Journal of Enzyme Inhibition and Medicinal Chemistry·Anjali Pandey, Leela Iyengar
Oct 7, 2020·Protein Science : a Publication of the Protein Society·Diane ChauliacKeelnatham T Shanmugam
Dec 6, 1994·Biochemistry·B N Leichus, J S Blanchard

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