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- Development of Bacillus methanolicus methanol dehydrogenase with improved formaldehyde reduction activity
- Jiyeun Yi; Jinhyuk Lee; Bong Hyun Sung; Du Kyeong Kang; Gyu Tae Lim; Jung Hoon Bae; Seung Goo Lee; S C Kim; Jung Hoon Sohn
- Bibliographic Citation
- Scientific Reports, vol. 8, pp. 12483-12483
- Publication Year
- 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 (kcat/Km) 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.
- Springer-Nature Pub Group
- Appears in Collections:
- Division of Biomedical Research > Disease Target Structure Research Center > 1. Journal Articles
Synthetic Biology and Bioengineering Research Institute > Synthetic Biology Research Center > 1. Journal Articles
Synthetic Biology and Bioengineering Research Institute > 1. Journal Articles
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