Modification of the monomer composition of polyhydroxyalkanoate synthesized in Saccharomyces cerevisiae expressing variants of the beta-oxidation-associated multifunctional enzyme.

Details

Serval ID
serval:BIB_F7D26095E568
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Modification of the monomer composition of polyhydroxyalkanoate synthesized in Saccharomyces cerevisiae expressing variants of the beta-oxidation-associated multifunctional enzyme.
Journal
Applied and Environmental Microbiology
Author(s)
Marchesini S., Erard N., Glumoff T., Hiltunen J.K., Poirier Y.
ISSN
0099-2240 (Print)
ISSN-L
0099-2240
Publication state
Published
Issued date
2003
Volume
69
Number
11
Pages
6495-6499
Language
english
Abstract
Expression by Saccharomyces cerevisiae of a polyhydroxyalkanoate (PHA) synthase modified at the carboxy end by the addition of a peroxisome targeting signal derived from the last 34 amino acids of the Brassica napus isocitrate lyase (ICL) and containing the terminal tripeptide Ser-Arg-Met resulted in the synthesis of PHA. The ability of the terminal peptide Ser-Arg-Met and of the 34-amino-acid peptide from the B. napus ICL to target foreign proteins to the peroxisome of S. cerevisiae was demonstrated with green fluorescent protein fusions. PHA synthesis was found to be dependent on the presence of both the enzymes generating the beta-oxidation intermediate 3-hydroxyacyl-coenzyme A (3-hydroxyacyl-[CoA]) and the peroxin-encoding PEX5 gene, demonstrating the requirement for a functional peroxisome and a beta-oxidation cycle for PHA synthesis. Using a variant of the S. cerevisiae beta-oxidation multifunctional enzyme with a mutation inactivating the B domain of the R-3-hydroxyacyl-CoA dehydrogenase, it was possible to modify the PHA monomer composition through an increase in the proportion of the short-chain monomers of five and six carbons.
Keywords
3-Hydroxyacyl CoA Dehydrogenases/genetics, Acyltransferases/genetics, Acyltransferases/metabolism, Enoyl-CoA Hydratase/genetics, Fatty Acids/metabolism, Gene Expression Regulation, Fungal, Genetic Engineering/methods, Multienzyme Complexes/genetics, Mutation, Oxidation-Reduction, Peroxisomes/enzymology, Polyesters/chemistry, Saccharomyces cerevisiae/enzymology, Saccharomyces cerevisiae/genetics
Pubmed
Web of science
Open Access
Yes
Create date
24/01/2008 19:42
Last modification date
20/08/2019 16:24
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