L-asparaginase (L-asparagine amidohydrolase, E.C.220.127.116.11) is a supplementary cellular enzyme used as an anticancer substance. 31 Streptomyces strains isolated from rhizosphere of some plants in different regions of Saudi Arabia. The highest 11 Streptomyces isolates in L-asparaginase activity were selected for further molecular analysis based on differences in 16S rRNA gene sequence. Alignment analysis using the BLAST tools confirmed that all isolates are related to Streptomyces with a similarity of 97% to 100%. And partial-length gene sequence was sentinto NCBI Gen Bank and received the accession numbers (Streptomyces griseoruben MN588164, Streptomyces alboflavus MN588165, Streptomyces werraensis MN588166, Streptomyces spororaveus MN588167, Streptomyces chryseus MN588168, Streptomyces globosus MN588169, Streptomyces roseolilacinus MN588170, Streptomyces xanthophaeus MN588171, Streptomyces lavendulae MN588172, Streptomyces tuirus MN588174, and Streptomyces tendae MN588175).