The biosynthetic pathway of polyunsaturated fatty acids (PUFAs) has been the subject of much interest over the last few years. Significant progress has been made in the identification of the enzymes required for PUFA synthesis; in particular, the fatty acid desaturases which are central to this pathway have now all been identified. These "front-end" desaturases are all members of the cytochrome b(5) fusion desaturase superfamily, since they contain an N-terminal domain that is orthologous to the microsomal cytochrome b(5). Examination of the primary sequence relationships between the various PUFA-specific cytochrome b(5) fusion desaturases and related fusion enzymes allows inferences regarding the evolution of this important enzyme class. More importantly, this knowledge helps underpin our understanding of polyunsaturated fatty acid biosynthesis.