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J Biotechnol. 2014 Dec 10;191:86-92. doi: 10.1016/j.jbiotec.2014.08.022. Epub 2014 Aug 26.

Biooxidation of n-butane to 1-butanol by engineered P450 monooxygenase under increased pressure.

Author information

1
Institute of Technical Biochemistry, Universitaet Stuttgart, Allmandring 31, 70569 Stuttgart, Germany(1).
2
BASF SE, 67056 Ludwigshafen, Germany.
3
Institute of Technical Biochemistry, Universitaet Stuttgart, Allmandring 31, 70569 Stuttgart, Germany(1). Electronic address: bernhard.hauer@uni-stuttgart.de.

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.

KEYWORDS:

1-Butanol; Butane hydroxylation; CYP153A; Fusion protein; High pressure bioconversion

PMID:
25169664
DOI:
10.1016/j.jbiotec.2014.08.022
[Indexed for MEDLINE]

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