title: Biooxidation of n-butane to 1-butanol by engineered P450 monooxygenase under increased pressure.
abstract: In addition to the traditional 1-butanol production by hydroformylation of gaseous propene and by fermentation of biomass, the cytochrome P450-catalyzed direct terminal oxidation of n-butane into the primary alcohol 1-butanol constitutes an alternative route to provide the high demand of this basic chemical. Moreover the use of n-butane offers an unexploited ubiquitous feed stock available in large quantities. Based on protein engineering of CYP153A from Polaromonas sp. JS666 and the improvement of the native redox system, a highly ω-regioselective (>96%) fusion protein variant (CYP153AP.sp.(G254A)-CPRBM3) for the conversion of n-butane into 1-butanol was developed. Maximum yield of 3.12g/L butanol, of which 2.99g/L comprise for 1-butanol, has been obtained after 20h reaction time. Due to the poor solubility of n-butane in an aqueous system, a high pressure reaction assembly was applied to increase the conversion. After optimization a maximum product content of 4.35g/L 1-butanol from a total amount of 4.53g/L butanol catalyzed by the self-sufficient fusion monooxygenase has been obtained at 15bar pressure. In comparison to the CYP153A wild type the 1-butanol concentration was enhanced fivefold using the engineered monooxygenase whole cell system by using the high-pressure reaction assembly.
authors.affiliation: Institute of Technical Biochemistry, Universitaet Stuttgart, Allmandring 31, 70569 Stuttgart, Germany(1).
authors.forename: Bernd A
authors.initials: BA
authors.lastname: Nebel
authors.affiliation: Institute of Technical Biochemistry, Universitaet Stuttgart, Allmandring 31, 70569 Stuttgart, Germany(1).
authors.forename: Daniel
authors.initials: D
authors.lastname: Scheps
authors.affiliation: Institute of Technical Biochemistry, Universitaet Stuttgart, Allmandring 31, 70569 Stuttgart, Germany(1).
authors.forename: Sumire
authors.initials: S
authors.lastname: Honda Malca
authors.affiliation: Institute of Technical Biochemistry, Universitaet Stuttgart, Allmandring 31, 70569 Stuttgart, Germany(1).
authors.forename: Bettina M
authors.initials: BM
authors.lastname: Nestl
authors.affiliation: BASF SE, 67056 Ludwigshafen, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Breuer
authors.affiliation: BASF SE, 67056 Ludwigshafen, Germany.
authors.forename: Hans-Günter
authors.initials: HG
authors.lastname: Wagner
authors.affiliation: BASF SE, 67056 Ludwigshafen, Germany.
authors.forename: Boris
authors.initials: B
authors.lastname: Breitscheidel
authors.affiliation: BASF SE, 67056 Ludwigshafen, Germany.
authors.forename: Detlef
authors.initials: D
authors.lastname: Kratz
authors.affiliation: Institute of Technical Biochemistry, Universitaet Stuttgart, Allmandring 31, 70569 Stuttgart, Germany(1). Electronic address: bernhard.hauer@uni-stuttgart.de.
authors.forename: Bernhard
authors.initials: B
authors.lastname: Hauer
mesh_terms: D020001:1-Butanol / Q000138:chemical synthesis* / Q000737:chemistry; D002073:Butanes / Q000138:chemical synthesis* / Q000737:chemistry; D003577:Cytochrome P-450 Enzyme System / Q000737:chemistry* / Q000235:genetics; D005285:Fermentation; D010084:Oxidation-Reduction; D011312:Pressure; D015202:Protein Engineering
chemical_list: D002073:Butanes; C046888:butane; D020001:1-Butanol; D003577:Cytochrome P-450 Enzyme System; C435228:cytochrome P-450 CYP153 (Acinetobacter)
keywords: 1-Butanol; Butane hydroxylation; CYP153A; Fusion protein; High pressure bioconversion