L-ASPARAGINASE is an anti-neoplasm agent. This agent is classified as an enzyme compound. L-asparaginase is used in medicine as well as in the food industry. Microbial strains that are producing L-Asparaginase can be found in many habitats such as soil. The aim of this study was to find isolates producing L-asparaginase enzyme with high ability to inhibit cancer cells. Bacteria of the Actinobacter family were isolated from the soil samples around the roots of the plants in the actinomycet isolation medium. Then, the bacteria were isolated and extracted by screening in M9 medium. Isolates were identified using16S PCR sequencing method. Three isolates of ASP and B and C showed the highest enzyme production, which showed a molecular weight of 43 kDa and in terms of enzymatic activity112. 572 IU/ml, 99.706 IU/ml, 138.572 IU/ml respectively. These three isolates with sequence analysis was similar to Streptomyces flavoviridis and Streptomyces griseus and Streptomyces hypolyticus respectively. Isolate C showed good effects in inhibiting cell MCF-7, HepG-2. There are various isolates enzyme production power in nature and Actinobacter family has shown more ability to produce various enzymes and metabolites and the isolates isolated in this study are used for industrial enzyme production.