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A quantitative analytical method for valienone and its application in the evaluation of valienone production by a breakthrough microbial process

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Chinese Journal of Natural Medicines 2017年第10期15卷 794-800页

作者： CUI Li Karin Yanagi SHI Ting LIU Zhang-Min BAI Lin-Quan FENG Yan State Key Laboratory of Microbial Metabolism School of Life Science & Biotechnology and Joint International Research Laboratory of Metabolic & Developmental Sciences Shanghai Jiao Tong University Shanghai 200240 China

Valienone is a significant natural carbasugar member of the C7-cyclitol family as a valuable precursor for glycosidase inhibitor *** is an intermediate of validamycin A biosynthesis pathway and exhibits minimal accumulation in the fermentation broth of the natural Streptomyces producer.A quantitative analytical method is crucial for the development of a breakthrough microbial process overcoming the consumption of the natural metabolic *** present study was designed to develop a pre-column derivatization high-performance liquid chromatography method for quantification of valienone and to help establish a straightforward fermentation process for valienone production by metabolically engineered Streptomyces hygroscopicus *** was derivatized by 2,4-dinitrophenylhydrazine(DNPH)in 10 mmol×L^(-1) H_3PO_4 at 37℃ for 45 min and the derivatives were separated on Eclipse XDB-C18(5 μm,4.6 mm×150 mm)column at 30 °C eluted with 50% acetonitrile for 18 *** derivatives were detected by diode array detector at 380 nm and the configurations of the derivatives were determined by computational *** method was shown to be effective,sensitive,and *** linearity was found in the range of 5–2 000 μg×mL^(-1).The intra-and inter-day precisions were 1.1%–2.7% and 1.7%–2.2%,*** absolute recovery of the spiked samples was 97.2%–102.6%.To date,this is the first reversed-phase high-performance liquid chromatography detection method for valienone in microbial culture *** method successfully helped evaluate the valienone production capability of the engineered Streptomyces hygroscopicus 5008 and could be promising for C7-cyclitol profiling of different engineered mutants combined with the metabonomics methods.

关键词： Valienone carbasugar Microbial process Derivatization HPLC

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