Characterization and heterologous expression of three DGATs from oil palm (Elaeis guineensis) mesocarp in Saccharomyces cerevisiae

Abstract Oil palm (Elaeis guineensis) can accumulate up to 88% oil in fruit mesocarp. A previous transcriptome study of oil palm fruits indicated that genes coding for three diacylglycerol acyltransferases (DGATs), designated as EgDGAT1_3, EgDGAT2_2 and EgWS/DGAT_1 (according to Rosli et al., 2018) were highly expressed in mesocarp during oil accumulation. In the present study, the corresponding open reading frames were isolated, and characterized by heterologous expression in the mutant yeast H1246, which is devoid of neutral lipid synthesis. Expression of EgDGAT1_3 or EgDGAT2_2 could restore TAG synthesis, confirming that both proteins are true DGAT. In contrast, expression of EgWS/DGAT_1 resulted in the synthesis of fatty acid isoamyl esters (FAIEs) with saturated long-chain and very-long-chain fatty acids. In the presence of exogenously supplied fatty alcohols, EgWS/DGAT_1 was able to produce wax esters, indicating that EgWS/DGAT_1 codes for an acyltransferase with wax ester synthase but no DGAT activity. Finally, the complete wax ester biosynthetic pathway was reconstituted in yeast by coexpressing EgWS/DGAT_1 with a fatty acyl reductase from Tetrahymena thermophila. Altogether, our results characterized two novel DGATs from oil palm as well as a putative wax ester synthase that preferentially using medium chain fatty alcohols and saturated very-long chain fatty acids as substrates.