Identification and Functional Characterization of a Novel Δ12 Fatty Acid Desaturase Gene from Haematococcus pluvialis

The freshwater microalga Haematococcus pluvialis accumulates large amounts of fatty acids in response to adverse conditions. However, the key fatty acid desaturase genes in H. pluvialis remain unknown. In this study, we cloned and functionally characterized a Δ12 fatty acid desaturase gene, and designated it as Hp FAD2. The open reading frame of Hp FAD2 consisted of 1137 base pairs and encoded 378 amino acids. The deduced polypeptide showed 70% identity to other endoplasmic reticulum Δ12 fatty acid desaturases, whereas it had only 44% identity to plastid Δ12 fatty acid desaturases. The PSORT algorithm and phylogenetic analysis further confirmed its affiliation to the endoplasmic reticulum Δ12 fatty acid desaturases. Heterologous expression was performed in Saccharomyces cerevisiae cells transformed with the recombinant plasmid p YES2-Hp FAD2. Two additional fatty acids(C16:2 and C18:2) were detected in the yeast transformants. The results indicated Δ12 desaturation activity and substrate preference for C18:1 over C16:1. The transcriptional levels of H. pluvialis Hp FAD2 at different growth stages were measured by quantitative polymerase chain reaction(PCR), indicating that the Hp FAD2 transcriptional levels were significantly higher in red cells than those in green cells. Our study brings more insight into the fatty acid biosynthetic pathway of H. pluvialis.