聚乙二醇/硫酸铵双水相萃取猪胃蛋白酶工艺研究

Extraction technology of pepsin from swine by PEG/ (NH4) 2SO4 aqueous two-phase system

摘要: 采用聚乙二醇/硫酸铵PEG/（NH4）2SO4建立稳定的双水相体系以分离猪胃蛋白酶。通过上下相体积比（VR）、比活力（SA）、纯化因子（PF）、酶活回收率（η）和SDS-聚丙烯酰胺凝胶电泳（SDS-PAGE）分析测定萃取效果。结果表明,萃取猪胃蛋白酶的最佳配比体系为25%PEG1000:18%（NH4）2SO4,所萃取的猪胃蛋白酶酶活回收率为91.5%,SDS-PAGE结果显示,萃取的蛋白质和猪胃蛋白酶对照品分子量大小一致。

Abstract: The pepsin was extracted and purified by PEG/ (NH4) 2SO4 aqueous two-phase system (ATPS) from stomach of swine.Extraction technology of pepsin using a newly established ATPS was investigated according to volume ratio (VR) , purification factor (PF) , specific activity (SA) , protease activity recovery yield (η) and SDS-polyacrylamide gel electrophoresis (SDS-PAGE) .The optimal ATPS conditions for pepsin partitioning was 25% PEG1000:18% (NH4) 2SO4, which activity recovery yield reached 91.5%.SDS-PAGE indicated that ATPS fraction had a major protein which molecular weight (MW) was consistent with swine pepsin.