Overexpression, biochemical characterization, and anticancer activates of L-asparaginase from Bacillus subtilis

L-asparaginases convert L-asparagine into L-aspartate and ammonia. The L-asparaginase from Bacillus subtilis was cloned expressed in the E. coli strain BL21(DE3)pLysS current study. Using glutathione sepharose 4B column chromatography, enzyme uniformly purified 173.34 times, with a final specific activity of 1769.13 IU/mg protein yield 56.14%. isolated identified as 36 kDa polypeptide chain by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. immobilized placed on top Ca alginate beads. is quite stable retains majority its at 4 °C (74 percent). enzymatic structural characteristics free recombinant were studied. peaked after 30 min incubation pH 8.0 45 °C. After minutes 50 °C, showed peak 8.5. refined enzyme's amino acid makeup identified. An that heals leukemia, L-asparaginase, can be successfully mass-produced using this technique.