ABSTRACT
OBJECTIVE: This work aimed at evaluating the influence of organic solvents and stationary phases in the extraction with glass beads and chromatographic purification of carotenoids, especially torularhodin, from Sporobolomyces ruberrimus. RESULTS: The combinations of acetone:hexane (1:1 v/v) and acetone:ethyl ether (1:1 v/v) yielded 171.74 and 172.19 µg of total carotenoids.g of cells-1, respectively. The first blend resulted in the highest percent of cell lysis of 57.4%. Among different proportions of acetone:hexane, the 9:1 v/v mixture showed a significant difference (p < 0.05), resulting in a recovery of total carotenoids of 221.88 µg.g of cells-1. The purification of carotenoids was made by preparative chromatography and the yield of the silica-containing stationary phase was higher (24 µg torularhodin.g cells-1). The analyses of the purified fractions in thin layer chromatography and high performance liquid chromatography indicated that the purification of carotenoids, especially of torularhodin, was successfully performed. CONCLUSIONS: The combination of polar (acetone) and non-polar solvents (hexane) and the use of silica as stationary phase was efficient to recover and purify torularhodin from the intracellular pigments of Sporobolomyces ruberrimus.
Subject(s)
Basidiomycota/chemistry , Carotenoids/isolation & purification , Solvents/chemistry , Acetone/chemistry , Carotenoids/chemistry , Chromatography, Liquid , Hexanes/chemistryABSTRACT
Torularhodin and torulene are two widespread microbial carotenoids with relatively few studies, as compared to other nutraceutical carotenoids such as β-carotene, lycopene and astaxanthin. Several genera of microorganisms produce it in high concentration (up to 0.1% of the cell dry weight), probably as a protection against photooxidation and free radicals. These pigments, which differ by a terminal carboxylic group, have provitamin-A activity and, being red, have potential use as food and cosmetic color additives. Several factors affect the biosynthesis of these substances, including: the composition of culture media, light irradiation, which may enhance the carotenoid production up to 25% of the non-irradiated cultures, and temperature, which changes the carotenoid balance towards more of the acidic carotenoid (torularhodin) or the hydrocarbon (torulene). The biomass may be directly extracted using non polar solvents such as hexane or a hexane-acetone mixture, without need of cell disruption. Extensive purification is not needed for using the pigments as food or cosmetic additives, but it is still necessary to evaluate the bioactivity of the pigments in humans.