AbstractProsopis juliflora is a drought-resistant evergreen spiny tree that grows in semi-arid and arid tracts of tropical and sub-tropical regions of the world. Dry pods of P. juliflora are a rich source of carbon (40% total sugar) and nitrogen (15% of total nitrogen) and so can be considered as a good substrate for the microbial growth. The present study was mainly focused on the utilization of these pods for the production and statistical optimization of oxytetracycline (OTC) from Streptomyces rimosus NCIM 2213 under SSF. The spectral characterization and chemical color reactions of purified OTC by UV, FTIR, 1H NMR, 13C NMR, and HPLC revealed that the structure was homologous to a standard sample. A central composite design with 26 trails yielded the following critical values of supplements to be added to the dry pods: maltose (0.125 g/gds), Inoculum size (0.617 mL/gds), CaCO3 (0.0026 g/gds), and moisture content (74.87%) with the maximum OTC yield 5.02 mg/gds.