Discovery and biochemical characterization of a methanol dehydrogenase from Lysinibacillus xylanilyticus

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Title
Discovery and biochemical characterization of a methanol dehydrogenase from Lysinibacillus xylanilyticus
Author(s)
Jin-Young Lee; Sung-Hyun Park; So-Hyung Oh; Jin Ju Lee; Kil Koang Kwon; Su Jin Kim; Minjeong Choi; Eugene RhaHyewon LeeDae-Hee LeeBong Hyun Sung; Soo Jin Yeom; Seung Goo Lee
Bibliographic Citation
Frontiers in Bioengineering and Biotechnology, vol. 8, pp. 67-67
Publication Year
2020
Abstract
Bioconversion of C1 chemicals such as methane and methanol into higher carbon-chain chemicals has been widely studied. Methanol oxidation catalyzed by methanol dehydrogenase (Mdh) is one of the key steps in methanol utilization in bacterial methylotrophy. In bacteria, few NAD+-dependent Mdhs have been reported that convert methanol to formaldehyde. In this study, an uncharacterized Mdh gene from Lysinibacillus xylanilyticus (Lxmdh) was cloned and expressed in Escherichia coli. The maximum alcohol oxidation activity of the recombinant enzyme was observed at pH 9.5 and 55°C in the presence of 10 mM Mg2+. To improve oxidation activity, rational approach-based, site-directed mutagenesis of 16 residues in the putative active site and NAD+-binding region was performed. The mutations S101V, T141S, and A164F improved the enzyme's specific activity toward methanol compared to that of the wild-type enzyme. These mutants show a slightly higher turnover rate than that of wild-type, although their K M values were increased compared to that of wild-type. Consequently, according the kinetic results, S101, T141, and A164 positions may related to the catalytic activity in the active site for methanol dehydrogenation. It should be further studied other mutant variants with high activity for methanol. In conclusion, we characterized a new Lxmdh and its variants that may be potentially useful for the development of synthetic methylotrophy in the future.
Keyword
Lysinibacillus xylanilyticusmethanol dehydrogenasemethanol oxidationmethylotrophyrational enzyme engineering
ISSN
2296-4185
Publisher
Frontiers Media Sa
DOI
http://dx.doi.org/10.3389/fbioe.2020.00067
Type
Article
Appears in Collections:
Division of Biomaterials Research > Synthetic Biology and Bioengineering Research Center > 1. Journal Articles
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