Development of Bacillus methanolicus methanol dehydrogenase with improved formaldehyde reduction activity

Abstract Methanol dehydrogenase (MDH), an NAD+-dependent oxidoreductase, reversibly converts formaldehyde to methanol. This activity is a key step for both toxic formaldehyde elimination and methanol production in bacterial methylotrophy. We mutated decameric Bacillus methanolicus MDH by directed ev...

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Autores principales:	Jiyeun Yi, Jinhyuk Lee, Bong Hyun Sung, Du-Kyeong Kang, GyuTae Lim, Jung-Hoon Bae, Seung-Goo Lee, Sun Chang Kim, Jung-Hoon Sohn
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2018
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/2bea7df7aed04a27bf1f7afbbc10fee8
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spelling	oai:doaj.org-article:2bea7df7aed04a27bf1f7afbbc10fee82021-12-02T15:09:03ZDevelopment of Bacillus methanolicus methanol dehydrogenase with improved formaldehyde reduction activity10.1038/s41598-018-31001-82045-2322https://doaj.org/article/2bea7df7aed04a27bf1f7afbbc10fee82018-08-01T00:00:00Zhttps://doi.org/10.1038/s41598-018-31001-8https://doaj.org/toc/2045-2322Abstract Methanol dehydrogenase (MDH), an NAD+-dependent oxidoreductase, reversibly converts formaldehyde to methanol. This activity is a key step for both toxic formaldehyde elimination and methanol production in bacterial methylotrophy. We mutated decameric Bacillus methanolicus MDH by directed evolution and screened mutants for increased formaldehyde reduction activity in Escherichia coli. The mutant with the highest formaldehyde reduction activity had three amino acid substitutions: F213V, F289L, and F356S. To identify the individual contributions of these residues to the increased reduction activity, the activities of mutant variants were evaluated. F213V/F289L and F213V/F289L/F356S showed 25.3- and 52.8-fold higher catalytic efficiency (k cat/K m) than wild type MDH, respectively. In addition, they converted 5.9- and 6.4-fold more formaldehyde to methanol in vitro than the wild type enzyme. Computational modelling revealed that the three substituted residues were located at MDH oligomerization interfaces, and may influence oligomerization stability: F213V aids in dimer formation, and F289L and F356S in decamer formation. The substitutions may stabilise oligomerization, thereby increasing the formaldehyde reduction activity of MDH.Jiyeun YiJinhyuk LeeBong Hyun SungDu-Kyeong KangGyuTae LimJung-Hoon BaeSeung-Goo LeeSun Chang KimJung-Hoon SohnNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 8, Iss 1, Pp 1-8 (2018)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Jiyeun Yi Jinhyuk Lee Bong Hyun Sung Du-Kyeong Kang GyuTae Lim Jung-Hoon Bae Seung-Goo Lee Sun Chang Kim Jung-Hoon Sohn Development of Bacillus methanolicus methanol dehydrogenase with improved formaldehyde reduction activity
description	Abstract Methanol dehydrogenase (MDH), an NAD+-dependent oxidoreductase, reversibly converts formaldehyde to methanol. This activity is a key step for both toxic formaldehyde elimination and methanol production in bacterial methylotrophy. We mutated decameric Bacillus methanolicus MDH by directed evolution and screened mutants for increased formaldehyde reduction activity in Escherichia coli. The mutant with the highest formaldehyde reduction activity had three amino acid substitutions: F213V, F289L, and F356S. To identify the individual contributions of these residues to the increased reduction activity, the activities of mutant variants were evaluated. F213V/F289L and F213V/F289L/F356S showed 25.3- and 52.8-fold higher catalytic efficiency (k cat/K m) than wild type MDH, respectively. In addition, they converted 5.9- and 6.4-fold more formaldehyde to methanol in vitro than the wild type enzyme. Computational modelling revealed that the three substituted residues were located at MDH oligomerization interfaces, and may influence oligomerization stability: F213V aids in dimer formation, and F289L and F356S in decamer formation. The substitutions may stabilise oligomerization, thereby increasing the formaldehyde reduction activity of MDH.
format	article
author	Jiyeun Yi Jinhyuk Lee Bong Hyun Sung Du-Kyeong Kang GyuTae Lim Jung-Hoon Bae Seung-Goo Lee Sun Chang Kim Jung-Hoon Sohn
author_facet	Jiyeun Yi Jinhyuk Lee Bong Hyun Sung Du-Kyeong Kang GyuTae Lim Jung-Hoon Bae Seung-Goo Lee Sun Chang Kim Jung-Hoon Sohn
author_sort	Jiyeun Yi
title	Development of Bacillus methanolicus methanol dehydrogenase with improved formaldehyde reduction activity
title_short	Development of Bacillus methanolicus methanol dehydrogenase with improved formaldehyde reduction activity
title_full	Development of Bacillus methanolicus methanol dehydrogenase with improved formaldehyde reduction activity
title_fullStr	Development of Bacillus methanolicus methanol dehydrogenase with improved formaldehyde reduction activity
title_full_unstemmed	Development of Bacillus methanolicus methanol dehydrogenase with improved formaldehyde reduction activity
title_sort	development of bacillus methanolicus methanol dehydrogenase with improved formaldehyde reduction activity
publisher	Nature Portfolio
publishDate	2018
url	https://doaj.org/article/2bea7df7aed04a27bf1f7afbbc10fee8
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Development of Bacillus methanolicus methanol dehydrogenase with improved formaldehyde reduction activity

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