响应曲面法优化蒜氨酸酶酶促反应条件

Optimization of Enzymatic Catalysis Conditions of Alliinase by Response Surface Methodology

以纯化的蒜氨酸酶为对象,研究影响蒜氨酸酶活性的不同因素并优化反应条件。在单因素试验的基础上,采用响应曲面法对影响蒜氨酸酶活性的4个主要因素包括酶与底物比、反应温度、pH值和离子浓度进行优化,以最大限度地提高酶促反应速率。Design-experts软件分析结果表明,蒜氨酸酶反应的最佳条件为:酶与底物比0.20、反应温度36℃、pH6.4、Mn2+浓度8.15mmol/L,该条件下蒜氨酸酶活力为(0.156±0.0053)U/mL。而Cu2+在10mmol/L的浓度条件下使蒜氨酸酶活力降低了一半左右。

The effects of different factors on the activity of alliinase purified from garlic cloves were investigated to optimize the reaction conditions of alliinase.On the basis of one-factor-at-a-time experiments,four factors affecting the activity of alliinase,including ratio,pH,reaction temperature and ion concentration,were optimized by response surface methodology to maximize the reaction rate.Analysis of the experimental data conducted using Design Expert software indicated that optimal parameters were ratio of 0.20,reaction temperature of 36 ℃,pH 6.4 and Mn2＋ concentration of 8.15 mmol/L.Under these optimal conditions,the activity of alliinase was（0.156 ± 0.0053） U/mL.However,Cu2＋ at the concentration of 10 mmol/L could reduce the initial activity of alliinase by 50%.