Construction and functional analysis of a whole-cell biocatalyst based on CYP108N7

- CYP108N7 was characterized as the first reported CYP108N subfamily member.
- CYP108N7 accepted surrogate redox partners.
- CYP108N7 was expressed together with redox partners and GDH in E.coli.
- CYP108N7 catalyzed epoxidation, hydroxylation, demethylation and dehalogenation.
- CYP108N7 catalyzed asymmetric sulfoxidation with excellent stereoselectivity.

Cytochrome P450 enzymes are versatile biocatalysts with great potential in biotechnology. A new bacterial P450 was identified from the genome of Rhodococcus wratislaviensis NBRC 100605 and annotated as CYP108N7. The enzyme accepted the ferredoxin and ferredoxin reductase from spinach as surrogate redox partners for improved electron transfer efficiency. It was heterologous expressed in Escherichia coli together with the redox partners and a glucose dehydrogenase which supplied the reduced cofactor NADPH. The resulting whole-cell biocatalyst catalyzed a variety of reactions including sulfoxidation, epoxidation, hydroxylation, demethylation and dehalogenation. Remarkable stereoselectivity was observed in asymmetric sulfoxidation reaction, which could deliver chiral sulfoxides with >99% ee from thioanisole and derivatives.