The Peroxisomal Acyl-CoA Thioesterase Pte1p from Saccharomyces cerevisiae Is Required for Efficient Degradation of Short Straight Chain and Branched Chain Fatty Acids

• Published in: The Journal of biological chemistry Vol. 281; no. 17; pp. 11729 - 11735
• Main Authors: Maeda, Isamu, Delessert, Syndie, Hasegawa, Seiko, Seto, Yoshiaki, Zuber, Sophie, Poirier, Yves
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 28.04.2006; American Society for Biochemistry and Molecular Biology
• Subjects: Acyl Coenzyme A - metabolism; Fatty Acids - chemistry; Fatty Acids - metabolism; Hydroxybutyrates - metabolism; Kinetics; Oxidation-Reduction; Peroxisomes - enzymology; Saccharomyces cerevisiae; Saccharomyces cerevisiae - metabolism; Thiolester Hydrolases - chemistry; Thiolester Hydrolases - genetics; Thiolester Hydrolases - metabolism
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.M511762200

The role of the Saccharomyces cerevisae peroxisomal acyl-coenzyme A (acyl-CoA) thioesterase (Pte1p) in fatty acid β-oxidation was studied by analyzing the in vitro kinetic activity of the purified protein as well as by measuring the carbon flux through the β-oxidation cycle in vivo using the synthesis of peroxisomal polyhydroxyalkanoate (PHA) from the polymerization of the 3-hydroxyacyl-CoAs as a marker. The amount of PHA synthesized from the degradation of 10-cis-heptadecenoic, tridecanoic, undecanoic, or nonanoic acids was equivalent or slightly reduced in the pte1Δ strain compared with wild type. In contrast, a strong reduction in PHA synthesized from heptanoic acid and 8-methyl-nonanoic acid was observed for the pte1Δ strain compared with wild type. The poor catabolism of 8-methyl-nonanoic acid via β-oxidation in pte1Δ negatively impacted the degradation of 10-cis-heptadecenoic acid and reduced the ability of the cells to efficiently grow in medium containing such fatty acids. An increase in the proportion of the short chain 3-hydroxyacid monomers was observed in PHA synthesized in pte1Δ cells grown on a variety of fatty acids, indicating a reduction in the metabolism of short chain acyl-CoAs in these cells. A purified histidine-tagged Pte1p showed high activity toward short and medium chain length acyl-CoAs, including butyryl-CoA, decanoyl-CoA and 8-methyl-nonanoyl-CoA. The kinetic parameters measured for the purified Pte1p fit well with the implication of this enzyme in the efficient metabolism of short straight and branched chain fatty acyl-CoAs by the β-oxidation cycle.