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

Abstract

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 A12 fatty acid desaturase gene, and designated it as HpFAD2. The open reading frame of HpFAD2 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 pYES2-HpFAD2. 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 HpFAD2 at different growth stages were measured by quantitative polymerase chain reaction (PCR), indicating that the HpFAD2 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.