聚乙二醇／硫酸铵双水相体系萃取猪胰蛋白酶.pdfVIP

学兔兔 化 工 进 展 2009年第28卷第2期 CHEMICAL INDUSTRY AND ENGD ERING PR0GRESS ·3O5· 聚乙二醇／硫酸铵双水相体系萃取猪胰蛋白酶 周红航，王维香 (西华大学生物工程学院，四川I成都 610039) 摘 要：采用聚乙二醇(PEG)，硫酸铵[(NH4)2SO4】双水相体系对猪胰蛋白酶分离进行了研究。通过综合考察酶分配 系数、蛋白质分配系数、相比和回收率，探讨了PEG400质量分数、(NH4)2s04质量分数、NaCI质量分数以及pH 值对胰蛋白酶萃取的影响，并通过正交实验进一步优化实验条件，结果表明(NH4)2SO4质量分数和PEG浓度对胰 蛋白酶的萃取影响大，在PEG400质量分数为24％、(NH4)2s04质量分数为21％、pH值为4．2所组成的双水相体 系下，可获得酶的高分配系数8．48，提取的胰蛋白酶活力达到1780U／mL。 关键词：胰蛋白酶；双水相萃取系统；聚乙二醇；分配系数 中图分类号：TQ 028．3 8 文献标识码：A 文章编号：1000—6613(2009)02—0305—05 Separation of porcine pancreatic trypsin using aqueous two-phase systems ZHOUHonghang，WANG Weixiang (School of Bioengineering Xihua University，Chengdu 610039，Sichuan，China) Abstract-Trypsin is an important proteolytic enzyme，already used in large—scale processes by the detergent and dairy industries．Separation of trypsin directly from porcine pancreatic homogenate in aqueous two—phase systems(ATPS)of polyethylene glycol／ammonium sulfate was studied．The effects of polyethylene glycol 400(PEG400)concentration，(NH4)2SO4 concentration，NaC1 concentration，and pH value on the extraction of porcine pancreatic trypsin were investigated in terms of phase volume， partition coefficients of both enzyme activity and protein，and recovery．Orthogonal experiment was used to further analyze and optimize the separation of trypsin．The results indicated that the control of the concentration of(NH4)2SO4 and PEG400 were the most important for partitioning of trypsin．The NaCI addition had little effect on trypsin partition．Maximum partition coefficient of