橄榄油乳化法测定脂肪酶活性的优化研究_滕宏飞.pdf

工艺技术 橄榄油乳化法测定脂肪酶活性的优化研究 滕宏飞，王丹静，徐青，辛建美，罗红宇* 浙江海洋学院食品与药学学院（舟山 316004） 摘 要 以猪胰脂肪酶为例，针对传统橄榄油乳化法存在准确度较低、重现性不佳等问题，对底物 浓度及用量、均质时间、缓冲液添加量、反应体系温度、反应环境等反应条件进行了优化研究：以 20%橄榄油乳化液5 mL代替25%橄榄油乳化液4 mL ，以3 mL磷酸盐缓冲液代替5 mL磷酸盐缓冲液， 将均质处理的速度与时间由高速(10 000 r/min)、6 min降低为5 000 r/min、2 min ，将反应体系温度 由40 ℃降低为35 ℃，改变静止的反应环境为在100 r/min下振荡反应，选择在空白组反应体系中加 入1 mL0.025 mol/L pH7.5磷酸缓冲液代替1 mL酶液。优化后的条件下测得的脂肪酶活力较优化前提 高了35% 以上，平行测定的RSD值为5.5%，方法的精密度较高。 关键词 橄榄油；乳化法；脂肪酶；优化 Optimization Study of Olive oil Emulsifi cation Method Determining Lipase Activity Teng Hong-fei,Wang Dan-jing, Xu Qing, Xin Jian-mei, Luo Hong-yu* Academy Of Food and Pharmacy, Zhejiang Ocean University (Zhoushan 316004) Abstract Taking porcine pancreas lipase for example, as the traditional olive oil emulsifi cation methods is with problems of low accuracy and poor reproducibility, the reaction conditions including the substrate concentration and amount, the homogenization time, the amount of buffer liquid added, reaction temperature and reaction environment were optimized. The 4 mL, 25% olive oil emulsion instead of 5 mL, 20% olive oil emulsion was used and the 3 mL phosphate buffer instead of 5 mL phosphate buffer was used. Meanwhile, the homogenization speed and time from high-speed ( 10 000 r/min), 6 min reduced to 5 000 r/min, 2 min; the reaction temperature decreased from 40 ℃ to 35 ℃; the static reaction environment was changed to 100 r/min oscillation; 1 mL, 0.025mol/L phosphate buffe at pH7.5 was added in the control group instead of 1 mL enzyme solution. Under optimum conditions, lipase activity is increased by 35% than the activity of before optimization and parallel determination of the RSD is 5.5%. The method is of high precision.