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Deconstruction of the CYP153A6 Alkane Hydroxylase System: Limitations and Optimization of In Vitro Alkane Hydroxylation

Catalysts. 2018-08; 
Svenja Kochius, Jacqueline Van Marwijk, Ana Cristina Ebrecht, Diederik Johannes Opperman and Martha Sophia Smit
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Bacterial Expression System The complete open reading frames of both the ferredoxin (Fdx) and ferredoxin reductase were codon optimized and synthesized by Genscript (Piscataway, NJ, USA) Get A Quote

摘要

Some of the most promising results for bacterial alkane hydroxylation to alcohols have been obtained with the cytochrome P450 monooxygenase CYP153A6. CYP153A6 belongs to the class I CYPs and is generally expressed from an operon that also encodes the ferredoxin (Fdx) and ferredoxin reductase (FdR) which transfer electrons to CYP153A6. In this study, purified enzymes (CYP, Fdx, FdR and dehydrogenases for cofactor regeneration) were used to deconstruct the CYP153A6 system into its separate components, to investigate the factors limiting octane hydroxylation in vitro. Proteins in the cytoplasm (cell-free extract) were found to better enhance and stabilize hydroxylase activity compared to bovine serum albumin (BSA)... More

关键词

cytochrome P450 monooxygenase; biocatalysis; CYP153A6; alkane hydroxylation; cell-free extract biotransformations; 1-octanol; cofactor regeneration