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Synthesis of chiral 2‐alkanols from n ‐alkanes by a P. putida whole‐cell biocatalyst
Author(s) -
Tieves Florian,
Erenburg Isabelle N.,
Mahmoud Osama,
Urlacher Vlada B.
Publication year - 2016
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.25953
Subject(s) - pseudomonas putida , biocatalysis , chemistry , octane , monooxygenase , hydroxylation , stereoselectivity , alkane , alcohol dehydrogenase , nonane , enantiomeric excess , stereochemistry , organic chemistry , alcohol , cytochrome p450 , enantioselective synthesis , enzyme , catalysis , reaction mechanism
The cytochrome P450 monooxygenase CYP154A8 from Nocardia farcinica was previously found to catalyze hydroxylation of linear alkanes (C 7 –C 9 ) 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.