A Rapid and Specific Method to Screen Epothilone High-Producing Strain with Spectrometry and Its Application

摘要

Epothilones are a kind of poliketide macrolide with strong stabilizing activities on polymerized microtubules which mimicked taxol. Microbial fermentation of Sorangium cellulosum is still the main way to produce epothilones with strain continuously improving by mutation. In general, yields of epothilones were detected by HPLC or LC-MS, which were time-consuming and not suited for high-throughput screening. In this paper, we described an efficient high-throughput method for epothilones high-producing strains with 96-well micro titer plate spectrometry combined with CCl_4 extracting. In this method, the purity of epothilones was 22 times higher than methanol extract. So the OD_(249) was much less disturbed by the impurities. Stabilization experiment and SPSS analysis of the relationship between the yields of epothilones detected by HPLC and the OD_(249) of the CCl_4 extracts of 192 mutated strains showed that our method was practical, accurate, and much more efficient.