A yellow pigment producing potent strain Streptomyces parvulus C5-5Y (KJ624998) isolated from leather industry was used in this study. The pigmented metabolite exhibited good spectrum of antagonistic activity against common human clinical pathogens. Hence, this work has been focused to analyze the bioactive compound present in the crude pigment. To achieve this, an initial step has been taken to purify the bioactive compound. The TLC and HPTLC analysis was carried out to analyze the number of compounds present in the crude pigment extract. Through silica gel column chromatography, the bioactive fraction was separated and named as F5. This fraction was efficient in combating the pathogens even at minor concentrations as supported by the MIC assay.  The UV-Vis spectral analysis revealed the presence of phenoxazinone chromophores in their structures by their absorption maxima at 443nm. The mass of the compound was found to be 1256m/z and the results from LC-MS/MS data unveiled the possibility of the compound F5 to fall under actinomycin group of antibiotics. A further assessment has to be taken to confirm it through NMR analysis and to analyze the compound to be employed as therapeutics through in vitro and in vivo studies.