title: Evolution of acyl-ACP-thioesterases and β-ketoacyl-ACP-synthases revealed by protein-protein interactions.
abstract: The fatty acid synthase (FAS) is a conserved primary metabolic enzyme complex capable of tolerating cross-species engineering of domains for the development of modified and overproduced fatty acids. In eukaryotes, acyl-acyl carrier protein thioesterases (TEs) off-load mature cargo from the acyl carrier protein (ACP), and plants have developed TEs for short/medium-chain fatty acids. We showed that engineering plant TEs into the green microalgaChlamydomonas reinhardtiidoes not result in the predicted shift in fatty acid profile. Since fatty acid biosynthesis relies on substrate recognition and protein-protein interactions between the ACP and its partner enzymes, we hypothesized that plant TEs and algal ACP do not functionally interact. Phylogenetic analysis revealed major evolutionary differences between FAS enzymes, including TEs and ketoacyl synthases (KSs), in which the former is present only in some species, whereas the latter is present in all, and has a common ancestor. In line with these results, TEs appeared to be selective towards their ACP partners whereas KSs showed promiscuous behavior across bacterial, plant and algal species. Based on phylogenetic analyses,in silicodocking,in vitromechanistic crosslinking andin vivoalgal engineering, we propose that phylogeny can predict effective interactions between ACPs and partner enzymes.
authors.affiliation: Department of Chemistry and Biochemistry, University of California-San Diego, La Jolla, CA 92093-0358, USA.
authors.forename: Joris
authors.initials: J
authors.lastname: Beld
authors.affiliation: Department of Chemistry and Biochemistry, University of California-San Diego, La Jolla, CA 92093-0358, USA.
authors.forename: Jillian L
authors.initials: JL
authors.lastname: Blatti
authors.affiliation: Department of Chemistry and Biochemistry, University of California-San Diego, La Jolla, CA 92093-0358, USA.
authors.forename: Craig
authors.initials: C
authors.lastname: Behnke
authors.affiliation: Department of Chemistry and Biochemistry, University of California-San Diego, La Jolla, CA 92093-0358, USA.
authors.forename: Michael
authors.initials: M
authors.lastname: Mendez
authors.affiliation: Department of Chemistry and Biochemistry, University of California-San Diego, La Jolla, CA 92093-0358, USA.
authors.forename: Michael D
authors.initials: MD
authors.lastname: Burkart
keywords: Chlamydomonas reinhardtii; acyl carrier protein; algae; bacteria; evolution; fatty acid synthase
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