title: Chromogenic nitrophenolate-based substrates for light-driven hybrid P450 BM3 enzyme assay.
abstract: The incorporation of a p-nitrophenoxy moiety in substrates has enabled the development of colorimetric assays to rapidly screen for O-demethylation activity of P450 enzymes. For the light-driven hybrid P450 BM3 enzymes, where a Ru(II) photosensitizer powers the enzyme upon visible light irradiation, we have investigated a family of p-nitrophenoxy derivatives as useful chromogenic substrates compatible with the light-driven approach. The validation of this assay and its adaptability to a 96-well plate format will enable the screening of the next generation of hybrid P450 BM3 enzymes towards C-H bond functionalization of non-natural substrates.
authors.affiliation: San José State University, Department of Chemistry, One Washington Square, San José, CA 95192-0101, United States.
authors.forename: Quan
authors.initials: Q
authors.lastname: Lam
authors.affiliation: San José State University, Department of Chemistry, One Washington Square, San José, CA 95192-0101, United States.
authors.forename: Alejandro
authors.initials: A
authors.lastname: Cortez
authors.affiliation: San José State University, Department of Chemistry, One Washington Square, San José, CA 95192-0101, United States.
authors.forename: Thanh Truc
authors.initials: TT
authors.lastname: Nguyen
authors.affiliation: San José State University, Department of Chemistry, One Washington Square, San José, CA 95192-0101, United States. Electronic address: mallory.kato@gmail.com.
authors.forename: Mallory
authors.initials: M
authors.lastname: Kato
authors.affiliation: San José State University, Department of Chemistry, One Washington Square, San José, CA 95192-0101, United States. Electronic address: lionel.cheruzel@sjsu.edu.
authors.forename: Lionel
authors.initials: L
authors.lastname: Cheruzel
mesh_terms: D002863:Chromogenic Compounds / Q000737:chemistry* / Q000378:metabolism; D003577:Cytochrome P-450 Enzyme System / Q000737:chemistry / Q000378:metabolism*; D057075:Enzyme Assays; D008027:Light; D009596:Nitrophenols / Q000737:chemistry*; D010777:Photochemistry; D017433:Protein Structure, Secondary
chemical_list: D002863:Chromogenic Compounds; D009596:Nitrophenols; D003577:Cytochrome P-450 Enzyme System
keywords: Chromogenic substrates; Colorimetric assay; Hybrid P450 BM3 enzymes; Light-driven biocatalysis; p-Nitrophenoxy derivatives