Extraction of pigments from natural sources is gaining momentum due to many biological applications. The aim of our study was to extract and to identify the pigment produced by Streptomyces species isolated from saltpan soil samples. The pigment-producing isolate was characterized by molecular taxonomy, identified as Streptomyces species, and designated as Streptomyces sp. VITVSK1. The isolate produced green color pigmentation upon solid substrate fermentation using parboiled rice as a media for 7 days at 37°C. The pigment derivative was extracted using methanol as solvent and purified by silica gel column chromatography and preparative thin layer chromatography using chloroform: methanol as solvent system. The purified compound was identified as 2,5-di-tert-butyl-1,4-benzoquinone (DTBBQ) based on similarity index with reference compounds available in the mass spectra library, NIST. Structure of the pure compound was also elucidated by 1H and 13C nuclear magnetic resonance spectra. The compound DTBBQ showed 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity with IC50 value of 0.6 μg/mL. DTBBQ also showed antimicrobial activity with a zone of inhibition of 21 mm against Bacillus cereus. The results of the present study showed that Streptomyces sp. VITVSK1 could be a promising source for the production of biologically active quinone-based pigments. © 2013 The Korean Society for Applied Biological Chemistry.