胰激肽原酶生产尾料中胰蛋白酶的制备工艺研究  

Study on Preparation of Trypsin from Kallikrein Production Tails

作　　者：王晓桐[1] 于泉 戴小敏 韦利军[2] 李谦[1] 欧瑜[1] 

机构地区：[1]中国药科大学生命科学与技术学院,江苏南京210009 [2]常州千红生化制药股份有限公司,江苏常州213022

出　　处：《药物生物技术》2015年第4期312-315,共4页Pharmaceutical Biotechnology

摘　　要：利用胰激肽原酶生产尾料作为原料提取胰蛋白酶,采用单因素试验优化纯化工艺,并以胰蛋白酶蛋白含量和酶活力作为指标检验纯化效果,分别采用BCA法、紫外吸收法等对所得产品进行蛋白含量和酶活力检测。结果发现胰蛋白酶纯化的最佳条件为:SP强阳离子交换树脂,醋酸盐洗脱液体系,洗脱液p H值为5.5。在此条件下,蛋白含量为3.42 mg/m L;胰蛋白酶总活力为2.18×106U、比活力为6.91×104U/mg;回收率为96.7%,纯化倍数为11.7倍。经过该法纯化提取,可以获得高纯度和符合国家药品标准的产品,使有限的资源发挥最大的利用价值。Trypsin was extracted and isolated from kallikrein production tails. Purification process was optimized by single factor experiment. The protein content and enzyme activity of trypsin were used as the index to detect the efficiency of isolation and purification. The protein content and enzyme activity of the products were detected by BCA method and Ultraviolet absorption method respectively. The results showed that the best conditions for isolation and purification of trypsin were using SP strong cation exchange resin,acetate buffer,and the buffer p H value was 5. 5. Under this condition,the protein content of products was 3. 42 mg / m L; the total enzyme activity of products was 2. 18 × 106 U,specific activity was 6. 91 × 104 U / mg,the rate of recovery was 96. 7% and the purification fold was 11. 7. By using this method to extract and purify,we can obtain high purity products which were in line with national standard,so that the limited resources may achieve the maximum value.

关 键 词：胰蛋白酶 离子交换色谱 提取纯化 单因素试验 

分 类 号：TQ460.6[化学工程—制药化工]