Phospholipids (PLs) are naturally-occurring amphiphilic compounds, which can be used as emulsifiers, liposome components, and nutritional supplements in food, cosmetic, and pharmaceutical industries. Enzymatic modifications of natural PLs are of importance to enhance the quality of PLs as value-added products. This chapter reviews the advances in the enzymatic modification of PLs, including acyl group modification and head group modification. Acyl group modification can be performed using lipase- or phospholipase A-mediated transesterification or ester synthesis to introduce particular fatty acid residues. Head group modification is carried out by phospholipase D-catalyzed transphosphatidylation. This reaction enables introduction of a wide range of natural and unnatural functional molecules into the phospholipids, making it possible to prepare chemically defined structured PLs for diverse applications.