Several soil isolates from 1 g of soil sample were isolated and screened for the production of L-asparaginase. Primary screening was performed using rapid plate assay; dye indicator studies were conducted, and phenol red with 0.005% concentration was found to be optimum. The secondary screening was carried out using the Nesslerization method. The bacteria screened for L-asparaginase production with no glutaminase activity was identified as Bacillus subtilis. Crude L-asparaginase enzyme was partially purified 1.57 folds of purity and 110 U/mg of specific activity. The glutaminase-free L-asparaginase activity was also confirmed using LC-MS analysis. The presence of mass peaks at 147.0 in the reaction mixture suggested an absence of glutaminase activity. An optimized medium obtained comprised of Dextrose 1.5 g/L, K₂HPO₄ 1.2 g/L, L-asparagine 15 g/L, and Tryptone 5 g/L. The highest L-asparaginase activity was observed at 6.0 pH and 30 °C. Kinetic parameters associated with biomass and L-asparaginase production were also studied. The computed values were µ_m 0.104 h⁻¹, X_m 6g/L P₀ 1.7U/mL P_m 8.2 U/mL Y_X/S 4 g-cell/g-glucose µP_m 0.35 h⁻¹ q_p 5.46 U/g/h Y_P/x 13.6667 U/g-cell. The novel bacterial isolates showed promise as a potential glutaminase-free L-asparaginase producer, which can prove to be of industrial applications.

从1 g土壤样品中分离出几种土壤分离物，并筛选出L-天冬酰胺酶的产生。使用快速平板测定法进行初步筛选；进行了染料指示剂研究，发现浓度为0.005％的酚红是最佳的。二次筛选是使用Nesslerization方法进行的。没有谷氨酰胺酶活性而筛选产生L-天冬酰胺酶的细菌被鉴定为枯草芽孢杆菌。粗制的L-天冬酰胺酶部分纯化，纯度为1.57倍，比活性为110 U / mg。还使用LC-MS分析确认了无谷氨酰胺酶的L-天冬酰胺酶活性。反应混合物中在147.0处存在质量峰，表明不存在谷氨酰胺酶活性。获得的优化培养基包括葡萄糖1.5 g / L，K ₂ HPO ₄ 1.2 g / L，L-天冬酰胺15 g / L和胰蛋白5 5 g / L。在6.0 pH和30°C下观察到最高的L-天冬酰胺酶活性。还研究了与生物量和L-天冬酰胺酶生产相关的动力学参数。计算值为µ _m 0.104 h ^-1，X _m 6g / LP ₀ 1.7U / mL P _m8.2 U / mL Y _{X / S} 4 g-cell / g-葡萄糖µ P _m 0.35 h ^-1 q _p 5.46 U / g / h Y _{P / x} 13.6667 U / g-cell。新型细菌分离物显示出有望作为潜在的无谷氨酰胺酶的L-天冬酰胺酶生产商，可以证明其具有工业应用价值。