Abstract
The European perch (Perca fluviatilis) is a carnivorous freshwater fish able to metabolise polyunsaturated fatty acids (PUFA) into highly unsaturated fatty acids (HUFA). This makes it a potential candidate for sustainable aquaculture development. In this study, special attention is given to the fatty-acid elongase (ELOVL) family, one of the two enzymatic systems implied in the HUFA biosynthesis. Structural information on European perch enzyme converting PUFA into HUFA is obtained by both molecular cloning and in silico characterization of an ELOVL5-like elongase from P. fluviatilis (pfELOVL). The full-length cDNA sequence consists of a 885-base pair Open Reading Frame coding for a 294-amino acid protein. Phylogenetic analysis and sequence alignment with fish elongases predict the pfELOVL clusters within the ELOVL5 sub-group. The amino-acid sequence displays the typical ELOVL features: several transmembrane α helices (TMH), an endoplasmic reticulum (ER) retention signal, and four "conserved boxes" involved in the catalytic site. In addition, the topology analysis predicts a 7-TMH structure addressed in the ER membrane. A 3D model of the protein embedded in an ER-like membrane environment is also provided using de novo modelling and molecular dynamics. From docking studies, two putative enzyme-substrate-binding modes, including H bonds and CH-π interactions, emphasize the role of specific residues in the "conserved boxes".