Discovery of an Epidermal Stearoyl-Acyl Carrier Protein Thioesterase

Plant epicuticular, or surface, waxes are synthesized primarily, if not exclusively, by epidermal cells. The epicuticular wax constitutes almost 20% of the chloroform-extractable lipids in developing leek leaf and is derived predominantly from saturated fatty acids. The significant requirement for saturated fatty acids in epidermal tissues led us to investigate whether or not epidermal extracts have thioesterase activities that prefer saturated acyl-acyl carrier protein (ACP) substrates, rather than the 18:1-ACP more commonly hydrolyzed by total leaf extracts. Epidermal extracts from Brassica, pea, and leek exhibited higher activities toward saturated acyl-ACPs relative to 18:1-ACP when compared to total leaf or leaf parenchymal extracts. We identified and purified a stearoyl-ACP (18:0-ACP)-specific thioesterase from leek epidermal extracts which could be separated from 18:1-ACP thioesterase using hydroxyapatite chromatography. The stearoyl-ACP thioesterase exhibited a high preference for 18:0-ACP, having less than 10% of the 18:0-ACP hydrolyzing activity when presented with 18:1-ACP, 16:0-ACP, or 18:0-CoA substrates. The stearoyl-ACP thioesterase was predominantly, if not exclusively, expressed in epidermis and may play a role in generating the saturated fatty acid pool required for wax production.