title: Oxygenation cascade analysis in conversion of n-octane catalyzed by cytochrome P450 CYP102A3 mutants at the P331 site.
abstract: To improve the hydroxylation efficiency of cytochrome P450 CYP102A3 to substrate n-octane, a previously reported triple-mutant F88V/S188Q/A330V with altered regioselectivity was selected and site-saturation mutagenesis was performed at its P331 site, which is adjacent to one of the active sites A330. Using whole-cell biotransformations to analyze the created mutants, four better mutants P331A, P331T, P331S, and P331V were achieved and exhibited significantly improved conversion rates toward n-octane, which are 33%, 33%, 22% and 28%, respectively, whereas the activity of P331R was greatly reduced and P331K gave almost zero conversion of n-octane. Besides the main product octanols, different octanones and 1,7-octanediol were also detected for some of the mutants. The above results demonstrated that the P331 site of CYP102A3 also plays an important role in the n-octane oxidation and CYP102A3 is a functionally flexible biocatalyst that can be optimized for a variety of industrial applications.
authors.affiliation: Institute of Chemical and Engineering Sciences, Jurong Island, 627833 Singapore.
authors.forename: Songhe
authors.initials: S
authors.lastname: Wang
authors.affiliation: Institute of Chemical and Engineering Sciences, Jurong Island, 627833 Singapore.
authors.forename: Zhibin
authors.initials: Z
authors.lastname: Liu
mesh_terms: D001412:Bacillus subtilis / Q000201:enzymology* / Q000235:genetics; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics; D020134:Catalytic Domain; D003577:Cytochrome P-450 Enzyme System / Q000737:chemistry* / Q000235:genetics; D009154:Mutation; D009795:Octanes / Q000737:chemistry*; D010084:Oxidation-Reduction; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics
chemical_list: D001426:Bacterial Proteins; D009795:Octanes; D011994:Recombinant Proteins; D003577:Cytochrome P-450 Enzyme System; C445019:CYP102A3 protein, Bacillus subtilis; C026728:octane
keywords: cytochrome P450 monooxygenase; hydroxylation; n-octane; site-saturation mutagenesis