MJM383, a rare actinomycete sp. strain originated from Chinese soils, was isolated through an antimicrobial screening system. The analysis of 16S rDNA sequences and biochemical characterization determined the strain to belong to genus Kitasatospora. Both NMR and ESI mass data of its purified bioactive compounds revealed Kitasatospora sp. MJM383 to produce two antitumor agents, streptonigrin and oxopropaline G, which have been known to be produced from Streptomyces species. This is the first report to demonstrate the presence of antitumor agents produced by genus Kitasatospora.