A single desaturase gene from red yeast Sporidiobolus pararoseus is responsible for both four- and five-step dehydrogenation of phytoene

Abstract

Carotenoids are one of the most common classes of natural pigments widely occurring within organisms. These structurally diverse pigments are of great importance in different processes such as nutrition, vision, cellular growth and development. While found in various yeast strains, one of the best-studied carotenoid producer is the pigmented species Sporidiobolus pararoseus. However, the precise nature of the genes involved in the biosynthesis of carotenoids in this species remains unclear. Here, we cloned a cDNA copy of the phytoene desaturase gene crtI from Sporidiobolus pararoseus CGMCC 2.5280. The crtI full-length genomic DNA and cDNA are 2330bp and 1683bp, respectively. This gene encodes a 560-amino acid protein with a predicted molecular mass of 62.28 kDa and a pI of 7.27. Functional identification of the gene was performed using heterologous complementation detection in Escherichia coli. Our experimental findings indicate that the enzymatic conversion of phytoene to lycopene (fourth step product) and 3,4-didehydrolycopene (fifth step product) is catalyzed by this phytoene desaturase of S. pararoseus through consecutive dehydrogenation. Furthermore, our findings suggest that the crtI gene of S. pararoseus represents an alternative gene source for the reconstruction of carotenogenic pathways vital for the production of engineered carotenoids.