Engineering of versatile redox partner fusions that support monooxygenase activity of functionally diverse cytochrome P450s.

ABSTRACT: Most bacterial cytochrome P450 monooxygenases (P450s or CYPs) require two redox partner proteins for activity. To reduce complexity of the redox chain, the Bacillus subtilis flavodoxin YkuN (Y) was fused to the Escherichia coli flavodoxin reductase Fpr (R), and activity was tuned by placing flexible (GGGGS)_n or rigid ([E/L]PPPP)_n linkers (n = 1-5) in between. P-linker constructs typically outperformed their G-linker counterparts, with superior performance of YR-P5, which carries linker ([E/L]PPPP)₅. Molecular dynamics simulations demonstrated that ([E/L]PPPP)_n linkers are intrinsically rigid, whereas (GGGGS)_n linkers are highly flexible and biochemical experiments suggest a higher degree of separation between the fusion partners in cas

SUBMITTER: Bakkes PJ 

PROVIDER: S-EPMC5575160 | biostudies-literature | 2017 Aug

REPOSITORIES: biostudies-literature