Genome wide identification and functional characterization of two LC-PUFA biosynthesis elongase (elovl8) genes in rabbitfish (Siganus canaliculatus)

Abstract

Elongases of very long-chain fatty acids (Elovls) catalyze the rate-limiting step of the elongation pathway that results in net 2‑carbon elongation of pre-existing fatty acyl chains. As a set of crucial enzymes involved in the long-chain polyunsaturated fatty acids (LC-PUFA) biosynthesis, Elovls of fish have been investigated extensively in recent years. In the present study, we first identified two novel fish-specific elovl genes (named as elovl8a and elovl8b) from the herbivorous marine teleost rabbitfish (Siganus canaliculatus) by genomic survey and molecular cloning methods. Subsequently, their functional characteristics, tissue distribution patterns and transcriptional changes in response to different nutritional states were investigated. Full-length coding sequences of the elovl8a and elovl8b genes were 804 and 792 bp, encoding 267 and 263 amino acids, respectively. Multiple alignment, genomic synteny and phylogenetic analyses further suggested that elovl8 genes were unique to teleosts. Functional characterization by heterologous expression in yeast showed that Elovl8b could elongate C18 (18:2n-6, 18:3n-3 and 18:4n-3) and C20 (20:4n-6 and 20:5n-3) polyunsaturated fatty acids (PUFA) to longer-chain polyunsaturated fatty acids (LC-PUFA) whereas Elovl8a lacked this ability. In vitro, the expression of elovl8b but not elovl8a in rabbitfish hepatocytes was significantly up-regulated by incubation with 18:2n-6, 18:3n-3, 20:4n-6 and 20:5n-3, respectively. In vivo, compared with fish oil, dietary vegetable oil enriched in C18 PUFA enhanced the expression of elovl8b in rabbitfish brain, liver, intestine and gill. These findings suggest that elovl8b but not elovl8a is a novel active member of the Elovl protein family involved in the LC-PUFA biosynthesis pathway in rabbitfish, and provide novel insight into the mechanisms of LC-PUFA biosynthesis in teleost.