Molecular Cloning and functional characterization of a putative Elovl4 gene and its expression in response to dietary fatty acid profiles in orange-spotted grouper Epinephelus coioides

Abstract

Elongase of very long-chain fatty acids (Elovl) 4 probably plays a crucial role in marine fish species, where lack of Elovl2 has been considered as one possible reason for their low long-chain polyunsaturated fatty acids' (LC-PUFAs) biosynthetic capability. Elongase of very long-chain fatty acids 4 is the most recent member of the Elovl family that has been investigated in fish. Here, we report the molecular cloning and functional characterization of putativeelovl4cDNA isolated from marine teleost,Epinephelus coioides, and its expression in response to dietary n-3 LC-PUFA and docosahexaenoic acid (DHA) to eicosapentaenoic acid (EPA) ratio. Theelovl4cDNA of grouper was 2341bp including 301bp of 5′-untranslated region (UTR), 918bp of the coding region that encodes 305 amino acids (AA) and 1122bp of 3′UTR. Heterologous expression in yeast demonstrated that grouper Elovl4 could elongate saturated fatty acids (FA), especially 24:0 and 26:0, up to 36:0. Also, grouper Elovl4 effectively converted C20 and C22 polyunsaturated FAs to elongated polyenoic products up to C36. Tissue distribution analysis revealed that Elovl4 were widely transcribed in various tissues with the highest level in eye, brain and testis as described in other teleosts. The transcript level ofelovl4was significantly affected by dietary n-3 LC-PUFA and high LC-PUFA level repressess its expression. However, the ratio of DHA to EPA had no significant influence on its expression. These results may contribute to better understanding the LC-PUFA biosynthetic pathway in this fish species.