响应面法优化α-淀粉酶的酶反应体系

目的:采用单因素实验及响应面法优化α-淀粉酶的反应体系。方法:以淀粉为底物对象，以可溶性淀粉浓度、α-淀粉酶浓度、反应时间为考察因素，在单因素实验基础上，运用Box-behnken实验设计方法研究各因素及其交互作用对α-淀粉酶作用底物时的反应速度的影响。结果:建立α-淀粉酶酶反应体系的最佳反应条件为12.0 mg/mL可溶性淀粉、1.50 U/mL α-淀粉酶、10.0 min反应时间，在此条件下，α-淀粉酶表现出的反应速度达到（19.53±1.74） mmol/（L·min），接近模型中的预测数值18.75 mmol/（L·min）。结论:此优化α-淀粉酶酶反应体系的方法可行，能够使α-淀粉酶在反应过程中发挥的酶活最大化，为日后在此体系下进行糖苷酶抑制剂的研究奠定了基础，具有一定的指导意义。

Optimization of Enzyme Reaction System of α-Amylase by Response Surface Methodology

Objective:To optimize the reaction system ofα-amylase by single-factor optimization and by response surface methodology.Methods:Based on single factor experiment using starch as substrate ofα-amylase,and regarding the soluble starch concentration,theα-amylase concentration and the reaction time as three major factors,the Box-behnken design experiment was used to optimize the reaction system conditions ofα-amylase and to study the influence of these factors and their interaction on the reaction rate ofα-amylase.Results:The optimizing system parameter were as follows:soluble starch 12.0 mg/mL,α-amylase 1.50 U/mL,and reaction time 10.0 min.Under the conditions,the reaction rate ofα-amylase was up to(19.53±1.74)mmol/(L·min).This result was near the predicted value of 18.75 mmol/(L·min)in the optimal model.Conclusion:The optimal method of reaction system ofα-amylase is reliable and practical significantly for makingα-amylase play an excellent activity,which to some extent lays the foundation for the study of glycosidase inhibitors efficiently under the optimal system.