Heliomycin (1) was isolated from the culture broth of actinomycete AB5. The structure of the isolated compound (1) was characterized by spectroscopic methods including NMR and mass spectrometry data. The influence of cultural parameters affecting the production of biomass including different culture media, pH values, temperatures, and incubation periods was investigated. The maximum production of secondary metabolites (2.78 g/l) was found in the culture medium of Waksman at initial pH 7.5 for four days at 35ºC. The antimicrobial activity of heliomycin (1) was evaluated against Gram-positive bacteria, Gram-negative bacteria, and fungi.