Contribution of CoA ligases to benzenoid biosynthesis in petunia flowers.

Abstract

Biosynthesis of benzoic acid from Phe requires shortening of the side chain by two carbons, which can occur via the β-oxidative or nonoxidative pathways. The first step in the β-oxidative pathway is cinnamoyl-CoA formation, likely catalyzed by a member of the 4-coumarate:CoA ligase (4CL) family that converts a range of trans-cinnamic acid derivatives into the corresponding CoA thioesters. Using a functional genomics approach, we identified two potential CoA-ligases from petunia (Petunia hybrida) petal-specific cDNA libraries. The cognate proteins share only 25% amino acid identity and are highly expressed in petunia corollas. Biochemical characterization of the recombinant proteins revealed that one of these proteins (Ph-4CL1) has broad substrate specificity and represents a bona fide 4CL, whereas the other is a cinnamate:CoA ligase (Ph-CNL). RNA interference suppression of Ph-4CL1 did not affect the petunia benzenoid scent profile, whereas downregulation of Ph-CNL resulted in a decrease in emission of benzylbenzoate, phenylethylbenzoate, and methylbenzoate. Green fluorescent protein localization studies revealed that the Ph-4CL1 protein is localized in the cytosol, whereas Ph-CNL is in peroxisomes. Our results indicate that subcellular compartmentalization of enzymes affects their involvement in the benzenoid network and provide evidence that cinnamoyl-CoA formation by Ph-CNL in the peroxisomes is the committed step in the β-oxidative pathway.

DOI: 10.1105/tpc.112.097519

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@article{Klempien2012ContributionOC, title={Contribution of CoA ligases to benzenoid biosynthesis in petunia flowers.}, author={Antje Klempien and Yasuhisa Kaminaga and Anthony V Qualley and Dinesh A. Nagegowda and Joshua R Widhalm and Irina Orlova and Ajit K. Shasany and Goro Taguchi and Christine M. Kish and Bruce R. Cooper and John Charles D'Auria and David Rhodes and Eran Pichersky and Natalia Dudareva}, journal={The Plant cell}, year={2012}, volume={24 5}, pages={2015-30} }