2034-56-2Relevant articles and documents
The CYPome of sorangium cellulosum so ce56 and identification of CYP109D1 as a new fatty acid hydroxylase
Khatri, Yogan,Hannemann, Frank,Ewen, Kerstin M.,Pistorius, Dominik,Perlova, Olena,Kagawa, Norio,Brachmann, Alexander O.,Mueller, Rolf,Bernhardt, Rita
experimental part, p. 1295 - 1305 (2011/09/20)
The first systematic study of the complete cytochrome P450 complement (CYPome) of Sorangium cellulosum So ce56, which is a producer of important secondary metabolites and has the largest bacterial genome sequenced to date, is presented. We describe the bioinformatic analysis of the So ce56 cytochrome P450 complement consisting of 21 putative P450 genes. Because fatty acids play a pivotal role during the complex life cycle of myxobacteria, we focused our studies on the characterization of fatty acid hydroxylases. Three novel potential fatty acid hydroxylases (CYP109D1, CYP264A1, and CYP266A1) were used for detailed characterization. One of them, CYP109D1 was able to perform subterminal hydroxylation of saturated fatty acids with the support of two autologous and one heterologous electron transfer system(s). The kinetic parameters for the product hydroxylation were derived.
Catalytic hydroxylation in biphasic systems using CYP102A1 mutants
Maurer, Steffen C.,Kuehnel, Katja,Kaysser, Leonard A.,Eiben, Sabine,Schmid, Rolf D.,Urlacher, Vlada B.
, p. 1090 - 1098 (2007/10/03)
Cytochrome P450 monooxygenases are biocatalysts that hydroxylate or epoxidise a wide range of hydrophobic organic substrates. Their technical application is, however, limited to a small number of whole-cell processes. The use of the isolated P450 enzymes is believed to be impractical due to their low stability, stoichiometric need of the expensive cofactor NAD(P)H and low solubility of most substrates in aqueous media. We investigated the behaviour of an isolated bacterial monooxygenase (mutants of CYP102A1) in a biphasic reaction system supported by cofactor recycling with the NADP +-dependent formate dehydrogenase from Pseudomonas sp 101. Using this experimental set-up cyclohexane, octane and myristic acid were hydroxylated. To reduce the process costs a novel NADH-dependent mutant of CYP102A1 was designed. For recycling of NADH an NAD+-dependent FDH was used. The stability of the monooxygenase mutants under the reaction conditions in the biphasic system was quite high as revealed by total turnover numbers of up to 12,850 in the NADPH-dependent cyclohexane hydroxylation and up to 30,000 in the NADH-dependent myristic acid oxidation.