An Actinomycetes producing an anti-VRSA (vancomycin-resistant Staphylococcus aureus) substance was isolated from soil. The cultural, morphological, physiological and phylogenetic analyses of an isolated strain were investigated for identification. Cultural characteristics based on ISP (International Streptomyces Project) were as follows: white aerial mycelium, yellow reverse side, and good growth on various medium. Also, the isolate did not produce the soluble pigment. Morphological characteristics were showed cylindrical spore chain and smooth spore surface by SEM (Scanning Electron Microscope). Physiological characteristics were showed LL-type by DAP isomer analysis and detected glycine, glutamic acid and alanine. A phylogenetic analysis of the 16S rDNA provided a clue that the isolated strain was actually a member of the genus Streptomyces, because the determined sequence exhibited a higher homology with Streptomyces echinatus. The isolate was identified to be a genus of Streptomyces sp.. The optimal culture conditions for the maximum production of anti-VRSA substance by Streptomyces sp. were attained in a culture medium composed of $2.0\%$ (w/v) glucose, and $0.4\%$ (w/v) yeast extract. The anti-VRSA substance was highly produced after 5 days of culture. Optimal pH and temperature conditions for the production of anti-VRSA substance were pH 7.0 and $28^{circ}C$, respectively.