Overexpression of a type 2 diacylglycerol acyltransferase in the marine diatom Phaeodactylum tricornutum enhances lipid production and omega-3 Long Chain Polyunsaturated Fatty Acid content
Abstract Background: Oleaginous microalgae represent a valuable resource for the production of high value molecules. Considering the importance of omega-3 long chain polyunsaturated fatty acids (LC-PUFAs) for human health and nutrition the yields of high value eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) require significant improvement to meet demand, however, the current cost of production remains high. A promising approach is to metabolically engineer strains with enhanced levels of triacylglycerols (TAGs) enriched in EPA and DHA. Results: Recently we have engineered the marine diatom Phaeodactylum tricornutum to accumulate enhanced levels of DHA in TAG. To further improve the incorporation of omega-3 LC-PUFAs in TAG we focused our effort on the identification of a type 2 acyl-CoA:diacylglycerol acyltransferase (DGAT) capable of improving lipid production and the incorporation of DHA in TAG. DGAT is a key enzyme in lipid synthesis. Following a diatom based in vivo screen of candidate DGATs, a native P. tricornutum DGAT2B was taken forward for detailed characterisation. Expression of DGAT2B in P. tricornutum was confirmed by qRT-PCR and the transgenic strain grew successfully in comparison to wildtype. PtDGAT2B has broad substrate specificity with preferences for C16 and LC-PUFAs acyl groups. Moreover, the expression of DGAT2B resulted in higher lipid yields and enhanced levels of DHA in TAG. Furthermore, co-expression with a Δ5-elongase showed how iterative metabolic engineering can be used to increase DHA and TAG content, irrespective of nitrogen treatment.Conclusion: This study provides further insight into lipid metabolism in P. tricornutum and suggests a metabolic engineering approach for the efficient production of EPA and DHA in microalgae.