鸭肉肌内磷脂水解酶的提取及相关酶系的酶活测定

Extraction of phospholipids lipolysis enzymatic in duck and determination of correlative enzymatic activity

为了优化肌内磷脂水解酶的提取过程,建立各种鸭肉肌内磷脂水解酶活的测定方法。采用pH 7.5、8.0、8.5的0.1 mol.L-1的Tris-HCl缓冲液提取鸭肉肌内磷脂水解酶,用饱和度范围为0～90%的硫酸铵进行盐析,研究新鲜鸭肉及板鸭中酸性脂肪酶、中性脂肪酶、总磷脂酶、磷脂酶A2(PLA2)、磷脂酶C(PLC)和磷脂酶D(PLD)的活力。结果表明,肌内磷脂水解酶的最佳pH为8.0,最佳盐析条件为70%饱和度的硫酸铵溶液。透析过后的粗酶液中酸性脂肪酶活略大于中性脂肪酶,磷脂酶活次之。

In order to opitimize the process of extracting phospholipase hydrolysis enzyme,assaying various phospholipase hydrolysis enzyme of the duck,0.1 mol·L-1 Tris-HCl buffer which pH was 7.5,8.0 and 8.5 which was used to extract the phospholipase hydrolysis enzyme of the duck and dry-cured duck respectively,ammonuium sulfate which had saturation ranged from 0 to 90% was adopted to precipitate the rude enzyme.What＇s more,the activity of acid lipase,neutral lipase,total phospholipase,phospholipase A2（PLA2）,phospholipase C（PLC） and phospholipase D（PLD） was studied.The results showed the most suitable pH for intramuscular phospholipid hydrolysis enzyme was 8.0 and 70% saturation of ammonuium sulfate was the most suitable precipitation condition.Moreover,the activity of acid lipase in dialysis solutions was the highest and the activity of total phospholipase was the third.