Bacterial acyl homoserine lactones triggered non‐native substrate hydroxylation catalysed by directed‐evolution‐derived cytochrome P450BM3 mutants DOI Creative Commons
Yuya Yokoyama, Shinya Ariyasu, Masayuki Karasawa

et al.

ChemCatChem, Journal Year: 2024, Volume and Issue: unknown

Published: Oct. 14, 2024

Abstract We report the directed evolution of cytochrome P450BM3 to efficiently utilize bacterial quorum sensing signalling molecule N ‐decanoyl homoserine lactone (C10‐HSL) as an effective decoy molecule. This represents first important step in our endeavor develop a self‐sufficient decoy‐molecule system whole‐cells that only necessitates addition culture medium and substrate realize hydroxylation non‐native substrates. Following five rounds evolution, mutant P450BM3, presence C10‐HSL, catalyzed benzene at rate 475 min −1 , highest turnover recorded for any P450 enzyme, achieving 46% yield whole‐cell reaction system. High‐resolution X‐ray crystal structure analysis series mutants narrates process, revealing how C10‐HSL is fixed binding pocket permit Finally, introduction synthase gene ExpI into Escherichia coli enabled situ production realizing, time, substrates without need laborious synthesis molecules.

Language: Английский

Bacterial acyl homoserine lactones triggered non‐native substrate hydroxylation catalysed by directed‐evolution‐derived cytochrome P450BM3 mutants DOI Creative Commons
Yuya Yokoyama, Shinya Ariyasu, Masayuki Karasawa

et al.

ChemCatChem, Journal Year: 2024, Volume and Issue: unknown

Published: Oct. 14, 2024

Abstract We report the directed evolution of cytochrome P450BM3 to efficiently utilize bacterial quorum sensing signalling molecule N ‐decanoyl homoserine lactone (C10‐HSL) as an effective decoy molecule. This represents first important step in our endeavor develop a self‐sufficient decoy‐molecule system whole‐cells that only necessitates addition culture medium and substrate realize hydroxylation non‐native substrates. Following five rounds evolution, mutant P450BM3, presence C10‐HSL, catalyzed benzene at rate 475 min −1 , highest turnover recorded for any P450 enzyme, achieving 46% yield whole‐cell reaction system. High‐resolution X‐ray crystal structure analysis series mutants narrates process, revealing how C10‐HSL is fixed binding pocket permit Finally, introduction synthase gene ExpI into Escherichia coli enabled situ production realizing, time, substrates without need laborious synthesis molecules.

Language: Английский

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