optimization of l-asparaginase type ii produced by bacillus velezensis se114

Once the acute lymphoblastic leukemia cells that need l-asparagine are exposed to l-asnase, they die because of the limitations of l-asparagine. the globally rising rate of all also requires extraordinary efforts to discover new microorganisms with high l-asnase production and efficiency. the aim of this study is the high amount of l-asnase production. after isolation, the l-asnase production was optimized using the response surface methodology and the central composite design. then, in-silico studies were predicted for the l-asnase-producing gene. in this study, bacillus velezensis was isolated as an l-asnase producer from slaughterhouse effluent using the m9 medium. the optimization process further illustrated tween 20, glucose, temperature, and l-asparagine, which were more significant for l-asnase production. based on statistical prediction by response surface methodology, more enzyme activity (7.11 u/ml) could be realized at 0.6% tween 20, 1.7% glucose, 55°c temperature, and 1.8% l-asparagine. the in-silico studies also established that the binding site is located at the n-terminal domain and the active site flexible loop. additionally, it contained thr36, ala47, tyr50, glu84, asp117, thr116, met142, lys189, thr193, and thr192 as the conserved and functional residues in l-asnase. it was concluded that b. velezensis se114  produced l-asnase type ii in the present study. the statistical optimization results also showed that tween 20, glucose, temperature, and l-asparagine were significant variables affecting the l-asnase production. in addition, temperature and l-asparagine had noteworthy interactions.