Nutritional and environmental conditions are important parameters that influence growth and secondary metabolite production by actinobacteria. Endophytic actinobacteria (n=283) isolated from different tissues (root, needle and stem) of Pinus roxburghii of Pauri (Garhwal) region, India, were screened for their antibacterial potential. A potent endophytic actinobacterial strain, GBTPR-167, isolated from root tissue of P. roxburghii was identified on the basis of morphological, chemotaxonomic and 16S rDNA analysis as Streptomyces sp. The strain was found to produce bioactive compound with high antibacterial activity in FM-2 medium. OFAT based optimization of factors effect the production of bioactive compound and maximum production was observed in medium containing glucose as carbon source and casein as nitrogen source at pH 7.0, 180 rpm and 30ºC on 7th day of incubation. Further, Streptomyces sp. displayed salt tolerance up to 6% NaCl concentration however; maximum antibacterial activity was obtained at 2% NaCl concentration. The study holds importance as enhanced production of antibacterial compound can be achieved with these improved parameters. Also, optimization strategies may be undertaken by using statistical tools like design expert.

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