食品科学 ›› 2012, Vol. 33 ›› Issue (17): 174-178.

• 生物工程 • 上一篇    下一篇

磷脂酶酶活测定条件的优化

占剑峰1,2,姜绍通1,2,*,潘丽军1,2   

  1. 1.安徽省农产品精深加工重点实验室 2.合肥工业大学生物与食品工程学院
  • 收稿日期:2011-09-14 修回日期:2012-07-23 出版日期:2012-09-15 发布日期:2012-11-09
  • 通讯作者: 姜绍通 E-mail:jiangshaotong@yahoo.com.cn
  • 基金资助:

    十二五国家支撑计划

Optimization of Determination Conditions for Phospholipase Activity

  • Received:2011-09-14 Revised:2012-07-23 Online:2012-09-15 Published:2012-11-09

摘要: 采用无油磷脂为底物,酸碱滴定法测定磷脂酶Lecitase甎ltra的酶活力,在单因素试验的基础上,选定反应时间、反应温度、反应pH值进行响应面分析试验,建立磷脂酶酶活力的二次回归方程,通过响应面分析得到酶活力测定的优化组合条件。结果表明:反应时间、反应温度、反应pH值对酶活力都有极其显著影响,测定酶活力的最佳条件为:底物质量浓度4g/100mL、终点pH 9.0、酶液稀释倍数100倍、反应pH5.1、反应温度53℃、反应时间8min,得到磷脂酶的酶活力为9550U/g。

关键词: 磷脂酶 Lecitase?Ultra, 酶活力测定, 酸碱滴定, 响应面分析

Abstract: To establish a method to determine the activity of phospholipase Lecitase?Ultra using oil-free phospholipids as the substrate, three reaction parameters including time, temperature and pH were optimized by response surface methodology based on one-factor-at-a-time experiments. A quadratic regression model for phospholipase activity was proposed. The results showed all these parameters had an extremely significant effect on phospholipase activity. The optimal conditions for determining phospholipase activity required substrate concentration of 4 g/100 mL, final pH 9.0, enzyme dilution factor of 100, reaction pH 5.1, reaction temperature of 53 ℃ and reaction time of 8 min.

Key words: phospholipase Lecitase?Ultra, enzyme activity determination, acid-base titration, response surface methodology