An attempt was made to enhance the bioactive metabolites by Nocardiopsis lucentensis VLK-104 under optimized conditions and evaluation of its bioactive metabolites by GC-MS analysis. Soil dilution plate technique was employed for the isolation of the strain on Starch-Casein agar (ISP-6) medium. Nocardiopsis lucentensis VLK-104 isolated from South Coast of Andhra Pradesh, India. The strain exhibited good antimicrobial activity against Gram-positive, Gram-negative and fungi. Five-day old culture exhibited maximum antimicrobial activity when cultured at pH 7.0 and temperature 30°C. Production of bioactive compounds by the strain varied significantly with different carbon and nitrogen sources. The productivity of the strain VLK-104 was enhanced by amending with carbon (Mannitol) and nitrogen (Peptone) sources @0.4% and 0.5% and K2HPO4(0.05%). As the strain exhibited good antimicrobial activity, it may be explored for biotechnological purposes. The crude ethyl acetate extract was analyzed by Gas Chromatography-Mass Spectroscopy. It reveals the metabolites produced by the strain and evidenced the presence of 41 diversified compounds according to the available library data, NIST MS Search (ver. 2.0). Hence it could be a possible source for investigate novel bioactive compounds by employing modern biotechnological aspects.