重组人Ⅱ型胰蛋白酶的性质及初步应用研究  

Research on Characteristic and Preliminary Application of Recombinant Human Trypsin Ⅱ

作　　者：纪德凯[1] 吴倩[1] 李素霞[1] 

机构地区：[1]华东理工大学生物反应器工程国家重点实验室,上海200237

出　　处：《食品与药品》2010年第11期384-388,共5页Food and Drug

摘　　要：目的研究重组人Ⅱ型胰蛋白酶的性质及初步应用。方法用米氏方程和双倒数作图法确定米氏常数Km,Vmax值,BAEE法测定胰蛋白酶活性,将胰蛋白酶用于激活羧肽酶原B及细胞消化。结果重组人Ⅱ型胰蛋白酶最适反应温度30℃,最适pH 7.6,以BAEE为底物,Km和Vmax分别为12.7μmol/L和212.76μmol/min。EDTA与PMSF抑制酶活性,高于50℃条件下易失活。与牛胰蛋白酶相比,重组人胰蛋白酶对羧肽酶原B有更高的激活效率,细胞消化方面二者效果基本相同。结论温度及pH是影响酶稳定性的较大因素,重组人胰蛋白酶可替代牛胰蛋白酶用于激活羧肽酶原B和细胞消化。Objective To study the characteristic and preliminary application of recombinant human trypsin II(rh-TII).Methods The Km and Vmax values were calculated using Michaelis equation and double-reciprocal plot method.The activity of trypsin was determined using BAEE as the substrate.The trypsin was used to activate procarboxypeptidase B and digest cells.Results The optimum reaction temperature and pH of rh-TII were 30 ℃ and 7.6,respectively.With BAEE as the substrate,the Km and Vmax values were 12.7 μmol/L and 2...

关 键 词：重组人Ⅱ型胰蛋白酶 牛胰蛋白酶 性质 应用 

分 类 号：Q789[生物学—分子生物学]