Urinary tract infection (UTI) is the most common and important clinical problem in millions of humans each year and drug resistant uropathogens like CTXM-extended spectrum β-lactamase (CTXM-ESBL) producers poses a catastrophic threat to healthcare community worldwide. ESBL pathogens were isolated from urine samples (90 nos) collected at Health center, VIT University, India. Phenotypic and genotypic characterization (CLSI-2010 guidelines) resulted inidentification 10 ESBLs isolates (Escherichiacoli(3 isolates)and Klebsiellapneumoniae (7 isolates).Uniplex PCR revealed CTXM-ESBLs (3/10). The Streptomyces species isolated from Cheyyur beach, Tamil Nadu, India, was screened for its inhibitory activity against clinically important MDR-ESBL producers (E.coli and K.pneumoniae). The ethyl acetate extract prepared fromStreptomycesspeciesexhibited anti-ESBL activity against CTXM-ESBL pathogens with the MIC value of 0.25 μg/mL against E. coli (S1), 2.0 μg/mL against K.pneumoniae (S2) and 0.5 μg/mL against E. coli (S3). Ertapenem was used as a positive control.The isolate was characterized by molecular taxonomy and identified to be belonged to the genus Streptomyces and designated as Streptomyces sp. VITSJK8.The results of our study shows that Streptomyces sp. VITSJK8 is the potential isolate for isolation anti- ESBL compounds.