Multiplexed activity metabolomics for isolation of filipin macrolides from a hypogean actinomycete
摘要
Chemical and biological stimulus screening in a hypogean actinomycete was used to elicit secondary metabolism. Optimal biosynthesis of bioactive natural products was identified using Multiplexed Activity Profiling for determining dose-dependent activity via six single-cell biological readouts. Bioactive extracts were fractioned to establish candidate compounds for isolation using Multiplexed Activity Metabolomics by correlating microtiter well-isolated phenotypes and extracted ion current peaks. This guided the isolation of four filipin polyene macrolides including a new metabolite filipin XV, an alkyl side-chain hydroxylated congener of the filipin chainin, with substantially attenuated cytotoxicity. Filipin-specific cytotoxicity was confirmed using flow cytometry and fluorescence microscopy.