PEG-柠檬酸盐双水相体系纯化α-淀粉酶及模型构建

Purification and modeling of α-amylase with aqueous two-phase systems

探索了采用PEG 柠檬酸盐双水相体系纯化α 淀粉酶的可行性和工艺条件 ,实验考察了PEG分子量 ,结线长度、pH、上清加入量和NaCl浓度对α 淀粉酶的分配的不同影响。双水相分离体系复杂 ,影响因素多 ,为进行分析、预测和优化 ,采用了响应面方法 ,以PEG 335 0浓度、柠檬酸盐浓度和NaCl浓度为自变量进行了α 淀粉酶分配系数、总蛋白分配系数、纯化因子和α 淀粉酶收率的优化 ,确定了特定的系统组成区域 ,通过实验验证了该区域内纯化因子大于 2 ,收率约 90 % 。

The feasibility of α amylase purification from fermentation supernatant of Bacillus subtilis with PEG citrate aqueous two phase system was studied PEG molecular weight,tie line length,supernatant amount,pH and NaCl addition had different effects on the partition of α amylase and a purification factor of 1 9 and 80% yield were reached Due to the complexity of aqueous two phase system,response surface methodology was employed to further analyze and optimize the purification of α amylase with the PEG 3 350,citrate and NaCl concentration as parameters Models of partition coefficients of α amylase and total protein,purification factor and α amylase yield were established and optimized and system composition area with purification factor of over 2 0 and 90% yield,as well as low system viscosity was determined and validated with experimental data