In this study, the secondary metabolites of a lichen-derived actinomycete strainAmycolatopsissp. YIM 130687 were investigated intensively by using three different media (4(#), 302(#), and 312(#)) for fermentation. A total of 21 compounds were isolated from the fermented extraction of the strain. The structures of all compounds were identified by the examination of HRESIMS and NMR spectra. Compounds1-3,5,6,21were only found in the cultivation on 302(#)medium, while compounds4,9-11were only obtained when the strain was cultured on 312(#)medium. On the other hand, compounds7,8, and20were only isolated from the fermentation product on 4(#)medium. The antimicrobial activity test showed that compound9had significant inhibitory effects on bacterial pathogens ofStaphylococcus aureusand MRSA with the MICs of 2 mu g/ml and fungal pathogens ofBotrytis cinereaandFusarium graminearumwith the MICs of 1 mu g/ml.