Separation And Determination Of Carotenoids In Sporidiobolus Pararoseus And Their Activities In Vitro

Carotenoids have been proved to have great advantages as natural pigments of food additives for their wide range of physiological functions and irreplaceable edible and medicinal value. We got a strain of Sporidiobolus pararoseus which might produce various kinds of carotenoids, such asβ-carotene,γ-carotene, torulene, torularhodin, and so on.The primary purpose of this subject was to research the activities of torulene and torularhodin under the conditions that there were only few papers on this aspects though the structures of them showed the same festures with lycopene. Torulene and torularhodin may be oxidant inhibitors or tumor suppressors.To confirm the categories and functions of the productions, a series of experiments were adopted in this study. n-Hexane and ethanol have been mixed to extact the pigments from fermentation products. TLC, HPLC and LC-ESI-MS were used to identification of these carotenoids after purification by silica gel column. The oxidation systems of DPPH (1,1- diphenyl-2- picrylhydrazyl radical 2,2- diphenyl-1-(2,4,6- trinitrophenyl) hydrazyl) radicals and linoleic acid and Hydrogen peroxide (H2O2) experiments were adopted to compare the antioxidant activities of the pigments we got. Antitumor effects experiments were comprised of morphological observation, MTT colorimetric, lactate dehydrogenase (LDH) experiments, cell cycle analysis, and hydrogen peroxide oxidation experiment.The main results are as follows:(1) The pigments in the fermentation of Sporidiobolus pararoseus were purified by silica gel column. The chromatographic characters of the pigments fit the features of carotenoids, and torulene and torularhodin were determinated.(2) Both of torulene and torularhodin have showed strong antioxidant abilities in the systems of DPPH radicals removing and linoleic acid antioxidant, especially torulene was stonger than lycopene. Hydrogen peroxide experiments evaluated the antioxidant properties of torulene and torularhodin in vitro on SK-HEP-1. In the range of this subject, roles of torulene or torularhodin which had been showed in pre- or post-treatment teams on SK-HEP-1 mode were not only antioxidantors, but synergetic damagers.(3) Torulene or torularhodin treatment could lead to morphological changes on SK-HEP-1. The MTT experiments results showed that the viabilities of SK-HEP-1 were inhibited by the treatment of torulene or torularhodin with a dose- and time-dependent pattern. The IC50 of torularhodin was 2.4μmol/L. There were significant differences between the effect of torularhodin and lycopene in SK-HEP-1 system.(4) Leaking of LDH had no obviously increased implied that the proliferation inhibition had no matter with cytotoxicity for the treatment of carotenoids. FCM analysis showed that higher concentration of carotenoids, more amounts of cells in G0/G1 phase, and lower the values of PI in these experiments.To conclude, torulene and torularhodin as the products of Sporidiobolus pararoseus showed strong antioxidant activity and antitumor effects in vitro culture cells SK-HEP-1 cell proliferation. The antitumor activity on SK-HEP-1 of torularhodin had been proved stronger than lycopene. The mechanism of antitumor was blocking the cell cycle to stay in G0/G1 phase. The PI value decreased.