beta-carotene and geranylgeranyl-pyrophosphate

Carotenoids are ubiquitous constituents of living organisms. These structurally diverse pigments have received considerable attention due to their biotechnological applications and potential beneficial effects on human health. In this study, we characterized an over-producing β-carotene mutant of Sporidiobolus pararoseus, obtained by ultraviolet mutagenesis, named MuY9. We compared the transcriptome between the wild-type and MuY9. A total of 348 differential expressed genes (DEGs) were found, and only one DEG crtYB is involved in carotenoid biosynthesis. The overproduction of β-carotene in MuY9 should be attributed to the up-regulation of crtYB. Functional identification of crtYB was performed using heterologous complementation in Escherichia coli. Our findings indicate that the enzymatic conversions of geranylgeranyl pyrophosphate to phytoene, as well as lycopene to β-carotene, are catalyzed by this CrtYB. Furthermore, our insights into the crtYB gene should facilitate a more detailed understanding of the carotenogenic pathway in S. pararoseus, and advance the development of the genetic engineering for the bio-production of carotenoids.

Topics: Basidiomycota; beta Carotene; Biosynthetic Pathways; Carotenoids; Escherichia coli; Fungal Proteins; Gene Expression Profiling; Gene Expression Regulation, Fungal; Genetic Complementation Test; Lycopene; Mutation; Phylogeny; Polyisoprenyl Phosphates

β-carotene has a wide range of application in food, pharmaceutical and cosmetic industries. For microbial production of β-carotene in Saccharomyces cerevisiae, the supply of geranylgeranyl diphosphate (GGPP) was firstly increased in S. cerevisiae BY4742 to obtain strain BY4742-T2 through over-expressing truncated 3-hydroxy-3-methylglutaryl-CoA reductase (tHMGR), which is the major rate-limiting enzyme in the mevalonate (MVA) pathway, and GGPP synthase (GGPS), which is a key enzyme in the diterpenoid synthetic pathway. The β-carotene synthetic genes of Pantoea agglomerans and Xanthophyllomyces dendrorhous were further integrated into strain BY4742-T2 for comparing β-carotene production. Over-expression of tHMGR and GGPS genes led to 26.0-fold increase of β-carotene production. In addition, genes from X. dendrorhous was more efficient than those from P. agglomerans for β-carotene production in S. cerevisiae. Strain BW02 was obtained which produced 1.56 mg/g (dry cell weight) β-carotene, which could be used further for constructing cell factories for β-carotene production.

Topics: Basidiomycota; beta Carotene; Farnesyltranstransferase; Hydroxymethylglutaryl CoA Reductases; Metabolic Engineering; Polyisoprenyl Phosphates; Saccharomyces cerevisiae