响应面法优化<i>α</i>-淀粉酶的酶反应体系

张程慧; 冯叙桥

doi:10.13386/j.issn1002-0306.2020040053

响应面法优化α-淀粉酶的酶反应体系

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

摘要

摘要: 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。

Abstract: Objective:To optimize the reaction system of α-amylase by single-factor optimization and by response surface methodology. Methods:Based on single factor experiment using starch as substrate of α-amylase,and regarding the soluble starch concentration,the α-amylase concentration and the reaction time as three major factors,the Box-behnken design experiment was used to optimize the reaction system conditions of α-amylase and to study the influence of these factors and their interaction on the reaction rate of α-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 of α-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 of α-amylase is reliable and practical significantly for making α-amylase play an excellent activity,which to some extent lays the foundation for the study of glycosidase inhibitors efficiently under the optimal system.