P450_BM3 fused to phosphite dehydrogenase allows phosphite-driven selective oxidations.

ABSTRACT: To facilitate the wider application of the NADPH-dependent P450_BM3, we fused the monooxygenase with a phosphite dehydrogenase (PTDH). The resulting monooxygenase-dehydrogenase fusion enzyme acts as a self-sufficient bifunctional catalyst, accepting phosphite as a cheap electron donor for the regeneration of NADPH.The well-expressed fusion enzyme was purified and analyzed in comparison to the parent enzymes. Using lauric acid as substrate for P450_BM3, it was found that the fusion enzyme had similar substrate affinity and hydroxylation selectivity while it displayed a significantly higher activity than the non-fused monooxygenase. Phosphite-driven conversions of lauric acid at restricted NADPH concentrations confirmed multiple turnovers of the cofactor. Interestingly, b

SUBMITTER: Beyer N 

PROVIDER: S-EPMC5320008 | biostudies-literature | 2017 Mar

REPOSITORIES: biostudies-literature