Biosensor-based directed evolution of methanol dehydrogenase from Lysinibacillus xylanilyticus

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dc.contributor.authorT K Le-
dc.contributor.authorJ B Ju-
dc.contributor.authorHyewon Lee-
dc.contributor.authorJin-Young Lee-
dc.contributor.authorSohyung Oh-
dc.contributor.authorKil Koang Kwon-
dc.contributor.authorBong Hyun Sung-
dc.contributor.authorSeung Goo Lee-
dc.contributor.authorS J Yeom-
dc.date.accessioned2021-02-09T03:30:35Z-
dc.date.available2021-02-09T03:30:35Z-
dc.date.issued2021-
dc.identifier.issn16616596-
dc.identifier.urihttps://oak.kribb.re.kr/handle/201005/24073-
dc.description.abstractMethanol dehydrogenase (Mdh), is a crucial enzyme for utilizing methane and methanol as carbon and energy sources in methylotrophy and synthetic methylotrophy. Engineering of Mdh, especially NAD-dependent Mdh, has thus been actively investigated to enhance methanol conversion. However, its poor catalytic activity and low methanol affinity limit its wider application. In this study, we applied a transcriptional factor-based biosensor for the direct evolution of Mdh from Lysinibacillus xylanilyticus (Lxmdh), which has a relatively high turnover rate and low KM value compared to other wild-type NAD-dependent Mdhs. A random mutant library of Lxmdh was constructed in Escherichia coli and was screened using formaldehyde-detectable biosensors by incubation with low methanol concentrations. Positive clones showing higher fluorescence were selected by fluorescence-activated cell sorting (FACS) system, and their catalytic activities toward methanol were evaluated. The successfully isolated mutants E396V, K318N, and K46E showed high activity, particularly at very low methanol concentrations. In kinetic analysis, mutant E396V, K318N, and K46E had superior methanol conversion efficiency, with 79-, 23-, and 3-fold improvements compared to the wild-type, respectively. These mutant enzymes could thus be useful for engineering synthetic methylotrophy and for enhancing methanol conversion to various useful products.-
dc.publisherMDPI-
dc.titleBiosensor-based directed evolution of methanol dehydrogenase from Lysinibacillus xylanilyticus-
dc.title.alternativeBiosensor-based directed evolution of methanol dehydrogenase from Lysinibacillus xylanilyticus-
dc.typeArticle-
dc.citation.titleInternational Journal of Molecular Sciences-
dc.citation.number3-
dc.citation.endPage1471-
dc.citation.startPage1471-
dc.citation.volume22-
dc.contributor.affiliatedAuthorHyewon Lee-
dc.contributor.affiliatedAuthorJin-Young Lee-
dc.contributor.affiliatedAuthorSohyung Oh-
dc.contributor.affiliatedAuthorKil Koang Kwon-
dc.contributor.affiliatedAuthorBong Hyun Sung-
dc.contributor.affiliatedAuthorSeung Goo Lee-
dc.contributor.alternativeNameLe-
dc.contributor.alternativeName주수빈-
dc.contributor.alternativeName이혜원-
dc.contributor.alternativeName이진영-
dc.contributor.alternativeName오소형-
dc.contributor.alternativeName권길광-
dc.contributor.alternativeName성봉현-
dc.contributor.alternativeName이승구-
dc.contributor.alternativeName염수진-
dc.identifier.bibliographicCitationInternational Journal of Molecular Sciences, vol. 22, no. 3, pp. 1471-1471-
dc.identifier.doi10.3390/ijms22031471-
dc.subject.keywordSynthetic methylotrophy-
dc.subject.keywordMethanol dehydrogenase-
dc.subject.keywordLysinibacillus xylanilyticus-
dc.subject.keywordBiosensor-
dc.subject.keywordScreening-
dc.subject.localLysinibacillus xylanilyticus-
dc.subject.localBiosensor-
dc.subject.localScreening-
dc.description.journalClassY-
Appears in Collections:
Division of Biomaterials Research > Synthetic Biology and Bioengineering Research Center > 1. Journal Articles
Division of Biomaterials Research > 1. Journal Articles
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