Thermotoga maritima TM0298 is a highly thermostable mannitol dehydrogenase

Applied Microbiology and Biotechnology
Seung Hoon SongClaire Vieille

Abstract

Thermotoga maritima TM0298 is annotated as an alcohol dehydrogenase, yet it shows high identity and similarity to mesophilic mannitol dehydrogenases. To investigate this enzyme further, its gene was cloned and expressed in Escherichia coli. The purified recombinant enzyme was most active on fructose and mannitol, making it the first known hyperthermophilic mannitol dehydrogenase. T. maritima mannitol dehydrogenase (TmMtDH) is optimally active between 90 and 100 degrees C and retains 63% of its activity at 120 degrees C but shows no detectable activity at room temperature. Its kinetic inactivation follows a first-order mechanism, with half-lives of 57 min at 80 degrees C and 6 min at 95 degrees C. Although TmMtDH has a higher V (max) with NADPH than with NADH, its catalytic efficiency is 2.2 times higher with NADH than with NADPH and 33 times higher with NAD(+) than with NADP(+). This cofactor specificity can be explained by the high density of negatively charged residues (Glu193, Asp195, and Glu196) downstream of the NAD(P) interaction site, the glycine motif. We demonstrate that TmMtDH contains a single catalytic zinc per subunit. Finally, we provide the first proof of concept that mannitol can be produced directly from glucos...Continue Reading

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Citations

Jul 7, 2009·Applied Microbiology and Biotechnology·Seung Hoon Song, Claire Vieille
Nov 11, 2010·Applied Microbiology and Biotechnology·Badal C Saha, F Michael Racine
Feb 25, 2011·Applied Microbiology and Biotechnology·Verena KallnikUwe Deppenmeier
Sep 29, 2011·Applied Microbiology and Biotechnology·Song-Yan HuangJian-Jiang Zhong
Aug 14, 2012·Journal of Bacteriology·Irina A RodionovaDmitry A Rodionov
Mar 26, 2014·Applied Microbiology and Biotechnology·Justin BeauchampClaire Vieille
Jan 23, 2015·Bioengineered·Justin Beauchamp, Claire Vieille
Oct 18, 2014·Applied Microbiology and Biotechnology·Isabel Oroz-GuineaEduardo García-Junceda
Dec 31, 2014·Applied and Environmental Microbiology·Agnès GroisillierThierry Tonon
Jan 1, 2013·ISRN Biotechnology·Sheelendra Mangal BhattSuresh Kumar Srivastava
Nov 26, 2020·Food Research International·Marwa Yagoub Farag KokoLi Yang

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