Structure-guided engineering of xylitol dehydrogenase cosubstrate specificity

Structure
Andreas H EhrensbergerDavid K Wilson

Abstract

Xylitol dehydrogenase (XDH) is one of several enzymes responsible for assimilating xylose into eukaryotic metabolism and is useful for fermentation of xylose contained in agricultural byproducts to produce ethanol. For efficient xylose utilization at high flux rates, cosubstrates should be recycled between the NAD+-specific XDH and the NADPH-preferring xylose reductase, another enzyme in the pathway. To understand and alter the cosubstrate specificity of XDH, we determined the crystal structure of the Gluconobacter oxydans holoenzyme to 1.9 angstroms resolution. The structure reveals that NAD+ specificity is largely conferred by Asp38, which interacts with the hydroxyls of the adenosine ribose. Met39 stacked under the purine ring and was also located near the 2' hydroxyl. Based on the location of these residues and on sequence alignments with related enzymes of various cosubstrate specificities, we constructed a double mutant (D38S/M39R) that was able to exclusively use NADP+, with no loss of activity.

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Citations

Apr 4, 2015·Enzyme and Microbial Technology·Ranjitha SinghJung-Kul Lee
Feb 4, 2010·Applied Microbiology and Biotechnology·Hee-Jung MoonJung-Kul Lee
Feb 24, 2010·Applied Microbiology and Biotechnology·Manish Kumar TiwariJung-Kul Lee
Jan 20, 2011·Applied Microbiology and Biotechnology·Ye-Wang ZhangJung-Kul Lee
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Jun 24, 2010·Physiological Genomics·Jonathan R ManningJohn J Mullins
Mar 2, 2018·Frontiers in Microbiology·Andrea M Chánique, Loreto P Parra

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