Glycosylphosphatidylinositol (GPI) anchoring is an important post-translational modification, which tethers non-transmembrane proteins to the outer leaflet of the plasma membrane (PM). It participates in many biological processes via facilitating signal perception, cell adhesion, transportation, and metabolism. Mature GPI moieties of eukaryotes usually contain a conserved glycan core structure and a variable lipid tail, and the lipid portion is important for targeting GPI-anchored proteins (GPI-APs) to the destined places at cell surface. The differences in lipid structure hence indicate varied sorting pathways for GPI-APs. GPI lipid synthesis involves multi-step remodeling reactions, which convert unsaturated fatty acid chains to saturated lipids and ultimately form varied lipid tails in yeast and animals.