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Phosphite‐driven Self‐sufficient Cytochrome P450
Author(s) -
Watanabe Hiroshi,
Hirakawa Hidehiko,
Nagamune Teruyuki
Publication year - 2013
Publication title -
chemcatchem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.497
H-Index - 106
eISSN - 1867-3899
pISSN - 1867-3880
DOI - 10.1002/cctc.201300445
Subject(s) - nad+ kinase , ferredoxin , chemistry , pseudomonas putida , dehydrogenase , biocatalysis , reagent , cytochrome , oxidizing agent , enzyme , combinatorial chemistry , catalysis , stereochemistry , biochemistry , organic chemistry , reaction mechanism
Cytochrome P450s (P450s) are attractive for biotechnology applications because P450s can catalyze regio‐ and stereo‐selective oxidations of non‐activated hydrocarbons. Many bacterial P450s require electron transfer from ferredoxins, which are in turn reduced by specific ferredoxin reductases and NAD(P)H. Stoichiometric consumption of NADH, an expensive reagent, is a major challenge for practical applications. A phosphite dehydrogenase (PTDH) that catalyzes the oxidation of phosphite to phosphate using NAD + , is an attractive choice of enzyme for NADH regeneration. In this study, we incorporated PTDH into an artificial heterotrimeric complex of Pseudomonas putida P450, putidaredoxin and putidaredoxin reductase, and the resulting complex almost completely hydroxylated 1 m m d ‐camphor using 10 m M phosphite in the presence of 20 μ M NAD + . This phosphite‐driven self‐sufficient P450 complex could provide a useful prototype for industry‐scale oxidizing biocatalysts.