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 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

SUBMITTER: Yi J 

PROVIDER: S-EPMC6102214 | biostudies-literature | 2018 Aug

REPOSITORIES: biostudies-literature