CYP94A5, a new cytochrome P450 from Nicotiana tabacum is able to catalyze the oxidation of fatty acids to the omega-alcohol and to the corresponding diacid.

摘要: A full length cDNA encoding a new cytochrome P450-dependent fatty acid hydroxylase (CYP94A5) was isolated from tobacco library. CYP94A5 expressed in S. cerevisiae strain WAT11 containing P450 reductase Arabidopsis thaliana necessary for catalytic activity of enzymes. When incubated 10 min presence NADPH with microsomes recombinant yeast, 9,10-epoxystearic converted into one major metabolite identified by GC/MS as 18-hydroxy-9,10-epoxystearic acid. The kinetic parameters the reaction were Km,app = 0.9 +/- 0.2 microM and Vmax,app 27 1 nmol x min(-1) nmol(-1) P450. Increasing incubation time to h led formation compound 9,10-epoxy-octadecan-1,18-dioic diacid also produced microsomal incubations Metabolites not yeast transformed control plasmid lacking their production inhibited antibodies raised against reductase, demonstrating involvement reactions. present study describes able catalyze complete set reactions oxidizing terminal methyl group corresponding carboxyl. This is enantioselective: after synthetic racemic mixture acid, chirality residual epoxide 40/60 favor 9R,10S enantiomer. catalyzed omega-hydroxylation saturated unsaturated acids aliphatic chain ranging C12 C18.