Triacylglycerol is the major component of vegetable oil, and acyl-CoA:diacylglycerol acyltransferase (DGAT) catalyzes the final step in its acyl-CoA dependent biosynthesis. In plants and microalgae, DGAT activity primarily resides in membrane-binding DGAT1 and DGAT2, with contributions from other DGATs such as bifunctional wax ester synthase/DGATs. This seminar will discuss the biochemical characterization of canola DGAT1, the generation of performance-enhanced DGAT1 variants through directed evolution, and their biotechnological potential. Additionally, the seminar will also discuss the characterization and improvement of microalgal DGATs, bridging knowledge gaps in microalgal lipid biosynthesis and biotechnology.