title: Synthesis of chiral 2-alkanols from n-alkanes by a P. putida whole-cell biocatalyst.
abstract: The cytochrome P450 monooxygenase CYP154A8 from Nocardia farcinica was previously found to catalyze hydroxylation of linear alkanes (C7 -C9 ) with a high regio- and stereoselectivity. The objective of this study was to integrate CYP154A8 along with suitable redox partners into a whole-cell system for the production of chiral 2-alkanols starting from alkanes. Both recombinant Escherichia coli and Pseudomonas putida whole-cell biocatalysts tested for this purpose showed the ability to produce chiral alkanols, but a solvent tolerant P. putida strain demonstrated several advantages in the applied biphasic reaction system. The optimized P. putida whole-cell system produced ∼16 mM (S)-2-octanol with 87% ee from octane, which is more than sevenfold higher than the previously described system with isolated enzymes. The achieved enantiopurity of the product could further be increased up to 99% ee by adding an alcohol dehydrogenase (ADH) to the alkane-oxidizing P. putida whole-cell systems. By using this setup for the individual conversions of heptane, octane or nonane, 2.6 mM (S)-2-heptanol with 91% ee, 5.4 mM (S)-2-octanol with 97% ee, or 5.5 mM (S)-2-nonanol with 97% ee were produced, respectively. The achieved concentrations of chiral 2-alkanols are the highest reported for a P450-based whole-cell system so far. Biotechnol. Bioeng. 2016;113: 1845-1852. © 2016 Wiley Periodicals, Inc.
authors.affiliation: Institute of Biochemistry, Heinrich-Heine University Düsseldorf, 40225, Düsseldorf, Germany.
authors.forename: Florian
authors.initials: F
authors.lastname: Tieves
authors.affiliation: Institute of Biochemistry, Heinrich-Heine University Düsseldorf, 40225, Düsseldorf, Germany.
authors.forename: Isabelle N
authors.initials: IN
authors.lastname: Erenburg
authors.affiliation: Institute of Biochemistry, Heinrich-Heine University Düsseldorf, 40225, Düsseldorf, Germany.
authors.forename: Osama
authors.initials: O
authors.lastname: Mahmoud
authors.affiliation: Institute of Biochemistry, Heinrich-Heine University Düsseldorf, 40225, Düsseldorf, Germany. vlada.urlacher@hhu.de.
authors.forename: Vlada B
authors.initials: VB
authors.lastname: Urlacher
mesh_terms: D000438:Alcohols / Q000032:analysis / Q000378:metabolism; D000473:Alkanes / Q000378:metabolism; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D060847:Metabolic Engineering / Q000379:methods*; D009615:Nocardia / Q000201:enzymology / Q000235:genetics; D000442:Octanols / Q000032:analysis / Q000378:metabolism*; D010084:Oxidation-Reduction; D016958:Pseudomonas putida / Q000235:genetics / Q000378:metabolism*; D013237:Stereoisomerism
chemical_list: D000438:Alcohols; D000473:Alkanes; D000442:Octanols; C067943:2-octanol; D003577:Cytochrome P-450 Enzyme System
keywords: 2-octanol; ADH; CYP154A8; P. putida; P450; alkanes