Semirationally Engineering an Efficient P450 Peroxygenase for Regio- and Enantioselective Hydroxylation of Steroids

ACS Catalysis, Journal Year: 2025, Volume and Issue: unknown, P. 2977 - 2986

Published: Feb. 5, 2025

Language: Английский

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Use of Whole Cells and Cell‐Free Extracts of Catalase‐Deficient E. coli for Peroxygenase‐Catalyzed Reactions

Biotechnology and Bioengineering, Journal Year: 2025, Volume and Issue: unknown

Published: Feb. 25, 2025

ABSTRACT Unspecific peroxygenases (UPOs) and cytochrome P450 monooxygenases (CYPs) with peroxygenase activity are becoming the preferred biocatalysts for oxyfunctionalization reactions. While whole cells (WCs) or cell‐free extracts (CFEs) of Escherichia coli often cofactor‐dependent monooxygenase reactions, hydrogen peroxide (H 2 O ) driven reactions generally performed purified enzymes, because catalases produced by E. expected to quickly degrade H . We used CRISPR/Cas system delete catalase encoding chromosomal genes, katG , katE from BL21‐Gold(DE3) obtain a catalase‐deficient strain. A short UPO, Dca two CYP peroxygenases, Ssca CYP_E284A CYP102A1_21B3, were compare strains expression subsequent sulfoxidation, epoxidation, benzylic hydroxylation activity. 10 mM was depleted within min after addition WCs CFEs wild‐type strain, at least 60% remained 24 h in lower turnover frequencies, benefited most use Comparison buffer versus strain revealed that as well proteins. also screen three SscaCYP, CYP_E284A, CYP_E284I against substrates comparing consumption substrate product formation. Finally, enzyme‐substrate pair highest activity, CYP_E284I, trans ‐β‐methylstyrene, preparative scale reaction WCs. Use instead enzymes can greatly benefit high‐throughput screening enzyme libraries while they be

Language: Английский

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Manfred T. Reetz – A Pioneer in Chemical Synthesis

Molecular Catalysis, Journal Year: 2024, Volume and Issue: 565, P. 114374 - 114374

Published: July 8, 2024

Language: Английский

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