Cytochrome P450 mutant

In a recent investigation to develop novel cytochrome P450 biocatalysts, DNA shuffling was used to produce chimeric cytochrome P450s mutants with enhanced biocatalytic activities, which were then co-expressed with NADPH-cytochrome reductase in E. coli to form an efficient system, in this case demonstrated to be effective for indole oxidation [69]. 

Scheme 10.8 Biosynthesis of epothilone. Individual PKS domains are represented as circles and individual NRPS domains as hexagons. Acyl carrier proteins (ACPs) and thiola-tion domains (T) are posttranslationally modified by a phos-phopantetheinyl group to which the biosynthetic intermediates are covalently bound throughout the chain assembly. The thioesterase domain (TE) cyclizes the fully assembled carbon chain to give the 16-membered lactone. Following dehydration of Cl 2—Cl 3 to give epothilones C and D, the final step in epothilone biosynthesis is the epoxidation of the C12=C13 double bond by the cytochrome P450 enzyme P450epol<. KS ketosyn-thase KS(Y) active-site tyrosine mutant of KS AT acyltransfer-ase C condensation domain A adenylation domain ...

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Harlow, G.R. and Halpert, J.R. (1998) Analysis of human cytochrome P450 3A4 cooperativity construction and characterization of a site-directed mutant that displays hyperbolic steroid hydroxylation kinetics. Proceedings of the National Academy of Sciences of the United States of America, 95 (12), 6636-6641. 

Kobayashi, Y., Fang, X., Szklarz, G. D., and Halpert, J. R. (1998) Probing the active site of cytochrome P450 2B1 metabolism of 7- alkoxycoumarins by the wild type and five site-directed mutants. Biochemistry 37, 6679-6688. 

OS158 Hong, Z., M. Ueguchi-Tanaka, K. Umemura, Set al. A rice brassino-steroids-deficient mutant, ebisu-dwarf (d2), is caused by a loss of function of a new member of cytochrome P450. Plant Cell 2003 15(12) 2900-2910. 

De vlieger, J., Krabbe, J. G., Commandeur, J. N. M., Vermeulen, N. P. E., Niessen, W. M. A., and Loftus, N. (2007). Characterisation of metabolites generated by mutant cytochromes P450 enzymes using a 3D ion trap-time of flight mass spectrometer. In Proceedings of the 55th ASMS Conference on Mass Spectrometry and Allied Topics. ASMS, Indianapolis, IN. 

Skoda RC, Gonzalez FJ, Demierre A, Meyer UA. Two mutant alleles of the human cytochrome P450 dbl gene (P450 II Dl) associated with genetically deficient metabolism of debrisoquine and other drugs. Proc Natl Acad Sci USA 1988 85 5240-5243. 

Melet A, Marques-Soares C, Schoch GA, et al. Analysis of human cytochrome P450 2C8 substrate specificity using a substrate pharmacophore and site-directed mutants. Biochemistry 2004 43 15379-15392. 

DeVoss JJ, Sibbesen O, Zhang ZP, et al. Substrate docking algorithms and prediction of the substrate specificity of cytochrome P450(cam) and its L244A mutant. J Am Chem Soc 1997 119 5489-5498. 

Khan KK, He YQ, Domanski TL, et al. Midazolam oxidation by cytochrome P450 3 A4 and active-site mutants an evaluation of multiple binding sites and of the metabolic pathway that leads to enzyme inactivation. Mol Pharmacol 2002 61(3) 495-506. 

Narasimhulu, S., and Willcox, J. K. (2001) Temperature-jump relaxation kinetics of substrate-induced spin-state transition in cytochrome P450 (Comparison of the wild-type and C334A mutant P450CAM and P4502B4), Arch. Bioch. and Bioph. 388, 198-206. 

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