| Popis: |
Publisher Summary This chapter describes procedures for identification of the carotenoid glycoside ester from Rhodococcus rhodochrous . Rhodococcus rhodochrous RNMS1 (IAM 13988) is cultured in a nutrient broth containing 1.0% (w/v) peptone, 0.3% (w/v) meat extract, and 0.2% (w/v) NaCl (pH 7.0) with shaking at 36 ° C for 45–60 hours. After the harvested wet cells (100–200 g) are dehydrated once with methanol by centrifugation, orange-colored material is extracted three or four times with chloroform-methanol (1:l, v/v) by centrifugation. The carotenoid extract is dissolved in a small volume of chloroform and diluted with hexane. It is submitted to silica gel 60 column chromatography and colorless materials are eluted with hexane. The monounsaturated esters are oxidized to diols with OsO 4 and the resulting dihydroxyl esters are converted into the corresponding trimethylsilyloxy derivatives. The principal fragmentation of the derivatives occurs between the trimethylsilyloxy-substituted carbon atoms, giving rise to a pair of intense fragments. |
