Biocatalyst engineering by assembly of fatty acid transport and oxidation activities for In vivo application of cytochrome P-450BM-3 monooxygenase

Details

Serval ID
serval:BIB_1EDDCDB2BD74
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Biocatalyst engineering by assembly of fatty acid transport and oxidation activities for In vivo application of cytochrome P-450BM-3 monooxygenase
Journal
Applied and Environmental Microbiology
Author(s)
Schneider  S., Wubbolts  M. G., Sanglard  D., Witholt  B.
ISSN
0099-2240 (Print)
Publication state
Published
Issued date
10/1998
Volume
64
Number
10
Pages
3784-90
Notes
Journal Article --- Old month value: Oct
Abstract
The application of whole cells containing cytochrome P-450BM-3 monooxygenase [EC 1.14.14.1] for the bioconversion of long-chain saturated fatty acids to omega-1, omega-2, and omega-3 hydroxy fatty acids was investigated. We utilized pentadecanoic acid and studied its conversion to a mixture of 12-, 13-, and 14-hydroxypentadecanoic acids by this monooxygenase. For this purpose, Escherichia coli recombinants containing plasmid pCYP102 producing the fatty acid monooxygenase cytochrome P-450BM-3 were used. To overcome inefficient uptake of pentadecanoic acid by intact E. coli cells, we made use of a cloned fatty acid uptake system from Pseudomonas oleovorans which, in contrast to the common FadL fatty acid uptake system of E. coli, does not require coupling by FadD (acyl-coenzyme A synthetase) of the imported fatty acid to coenzyme A. This system from P. oleovorans is encoded by a gene carried by plasmid pGEc47, which has been shown to effect facilitated uptake of oleic acid in E. coli W3110 (M. Nieboer, Ph.D. thesis, University of Groningen, Groningen, The Netherlands, 1996). By using a double recombinant of E. coli K27, which is a fadD mutant and therefore unable to consume substrates or products via the beta-oxidation cycle, a twofold increase in productivity was achieved. Applying cytochrome P-450BM-3 monooxygenase as a biocatalyst in whole cells does not require the exogenous addition of the costly cofactor NADPH. In combination with the coenzyme A-independent fatty acid uptake system from P. oleovorans, cytochrome P-450BM-3 recombinants appear to be useful alternatives to the enzymatic approach for the bioconversion of long-chain fatty acids to subterminal hydroxylated fatty acids.
Keywords
*Bacterial Proteins Biotechnology/methods Biotransformation Catalysis Cloning, Molecular Cytochrome P-450 Enzyme System/*metabolism Escherichia coli/*genetics Fatty Acids/metabolism Gas Chromatography-Mass Spectrometry Hydroxy Acids/*metabolism Kinetics Mixed Function Oxygenases/*metabolism Oxidation-Reduction Plasmids Recombinant Proteins/metabolism
Pubmed
Web of science
Create date
25/01/2008 14:40
Last modification date
20/08/2019 12:54
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