Abstract: Microbial secondary metabolites have a wide range of biological activities due to their structural diversity and have been proved a major source for drug lead compounds. However, the traditional method of a single culture restricts the metabolic pathways of microorganisms and as a result many metabolites cannot be formed. Recently, it has attracted much attention to use various techniques to activate those metabolic pathways restricted by the traditional method to get metabolic products with rich variety of structures. “One strain many compounds” (OSMAC) is a simple and effective approach for activating metabolic pathways and has been successfully applied. This review summarizes the common strategies of the OSMAC approach (altering cultivation parameters, co-cultivation, addition of enzyme inhibitors, etc）and the recent advances of OSMAC combined with genomics scanning. This review also introduces the research of our studies using the OSMAC approach on a fungus Spicaria elegans KLA03 which yielded a series of cytochalasins.