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Immobilization of Multi-biocatalysts in Alginate Beads for Cofactor Regeneration and Improved Reusability
Published on: April 22, 2016
Construction and functional analysis of a whole-cell biocatalyst based on CYP108N7
1Key Laboratory of Environmental and Applied Microbiology, Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu 610041, China; Environmental Microbiology Key Laboratory of Sichuan Province, Chengdu 610041, China; University of the Chinese Academy of Sciences, Beijing 100049, China.
Abstract:
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.
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