Co-cultivation is one of the ways to induce the discovery of new compounds by activating the cryptic genes in a microorganism’s genome. Actinomycetes is an order of bacteria with many clusters of gene coding for bioactive compounds that has not yet been expressed in the genome. In this study, co-cultivation between Actinomycetes strain DHE5-1 with pathogenic bacteria are conducted. Activity assay was done with disk diffusion method and the bioactive compounds in secondary metabolites are identified using HPLC UV-Vis. The results showed the activity of secondary metabolites from co-cultivation as an antimicrobial agent against the pathogenic microorganisms tested. The bioactive metabolite compounds showed different profiles produced between the mono and co-culture based on the models of the chromatogram. Finally, co-cultivation has the potential to induce the expression of covert genes in the organisms’ genome. This finding can promote the discovery of novel compounds with enhanced antimicrobial activity against resistant bacteria such as MDR-SA.