Expression of a type 2 diacylglycerol acyltransferase from Thalassiosira pseudonana in yeast leads to incorporation of docosahexaenoic acid β-oxidation intermediates into triacylglycerol.

Glycerolipids of the marine diatom Thalassiosira pseudonana are enriched particularly with eicosapentaenoic acid ( EPA), and also with an appreciable level of docosahexaenoic acid ( DHA). The present study describes the functional characterization of a type 2 diacylglycerol acyltransferase ( DGAT2, EC 2.3.1.20) from T. pseudonana, designated Tp DGAT2, which catalyzes the final step of triacylglycerol ( TAG) synthesis. Heterologous expression of this gene restored TAG formation in a yeast mutant devoid of TAG biosynthesis. Tp DGAT2 was also shown to exert a large impact on the fatty acid profile of TAG. Its expression caused a 10-12% increase of 18:1 and a concomitant decrease of 16:0 relative to that of At DGAT1( Arabidopsis thaliana). Furthermore, in the presence of the very-long-chain polyunsaturated fatty acids ( VLCPUFA) EPA and DHA, TAG formed by Tp DGAT2 displayed three- to six-fold increases in the percentage of VLCPUFA relative to that of At DGAT1 even though Tp DGAT2 conferred much lower TAG-synthetic activities than Arabidopsis DGAT1. Strikingly, when fed DHA, the yeast mutant expressing Tp DGAT2 incorporated high levels of EPA and DHA isomers derived from DHA β-oxidation. In contrast, no such phenomenon occurred in the cells expressing At DGAT1. These results suggested that, in addition to the role in breaking down storage lipids, yeast peroxisomes also contribute to lipid synthesis by recycling acyl-CoAs when a fatty acyl assembly system is available to capture and utilize the fatty acyl pools generated via β-oxidation. Our study hence illustrated a case where the efficiency and substrate preference of an acyltransferase can elicit far reaching metabolic adjustments that affect TAG composition. [ABSTRACT FROM AUTHOR]