title: Versatility of acyl-acyl carrier protein synthetases.
abstract: The acyl carrier protein (ACP) requires posttranslational modification with a 4'-phosphopantetheine arm for activity, and this thiol-terminated modification carries cargo between enzymes in ACP-dependent metabolic pathways. We show that acyl-ACP synthetases (AasSs) from different organisms are able to load even, odd, and unnatural fatty acids onto E. coli ACP in vitro. Vibrio harveyi AasS not only shows promiscuity for the acid substrate, but also is active upon various alternate carrier proteins. AasS activity also extends to functional activation in living organisms. We show that exogenously supplied carboxylic acids are loaded onto ACP and extended by the E. coli fatty acid synthase, including unnatural fatty acid analogs. These analogs are further integrated into cellular lipids. In vitro characterization of four different adenylate-forming enzymes allowed us to disambiguate CoA-ligases and AasSs, and further in vivo studies show the potential for functional application in other organisms.
authors.affiliation: Department of Chemistry and Biochemistry, University of California San Diego, 9500 Gilman Drive, 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, 9500 Gilman Drive, La Jolla, CA 92093-0358, USA.
authors.forename: Kara
authors.initials: K
authors.lastname: Finzel
authors.affiliation: Department of Chemistry and Biochemistry, University of California San Diego, 9500 Gilman Drive, La Jolla, CA 92093-0358, USA. Electronic address: mburkart@ucsd.edu.
authors.forename: Michael D
authors.initials: MD
authors.lastname: Burkart
mesh_terms: D017360:Arabidopsis / Q000201:enzymology; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D019730:Carbon-Sulfur Ligases / Q000235:genetics / Q000378:metabolism*; D003066:Coenzyme A Ligases / Q000378:metabolism; D004926:Escherichia coli / Q000378:metabolism; D005227:Fatty Acids / Q000737:chemistry / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D013379:Substrate Specificity; D046939:Synechocystis / Q000201:enzymology; D016963:Thermus thermophilus / Q000201:enzymology; D014733:Vibrio / Q000201:enzymology*
chemical_list: D001426:Bacterial Proteins; D005227:Fatty Acids; D011994:Recombinant Proteins; D019730:Carbon-Sulfur Ligases; D003066:Coenzyme A Ligases; C018749:long-chain-fatty-acid-(acyl-carrier-protein) ligase