Physical-Chemistry Department, Faculty of Biochemical and Pharmaceutical Sciences, National University of Rosario, FonCyT and CONICET, Suipacha 570, (S2002RLK) Rosario, Argentina.
J Chromatogr B Analyt Technol Biomed Life Sci. 2011 Apr 15;879(13-14):1003-7. doi: 10.1016/j.jchromb.2011.02.037. Epub 2011 Mar 1.
The condition of chymotrypsin (ChTRP)-Eudragit® (Eu) insoluble complex formation was studied with the aim of applying this information to the separation of chymotrypsin from a crude filtrate of bovine pancreas homogenate. The optimal pH of the complex precipitation was 4.60 for ChTRP-Eudragit® L100 and 5.40 for ChTRP-Eudragit® S100. The polyelectrolyte concentration necessary for the commercial enzyme precipitation was lower than 0.1% (w/v). The complex formation was inhibited by NaCl for both polyelectrolytes. The method was applied to recover the enzyme from bovine homogenate; ChTRP was precipitated by Eudragit® addition. The non-soluble complexes were separated by simple centrifugation and re-dissolved by a pH change to 8.20. The best conditions to recover ChTRP brought about a purification factor of around 4 and 90% yield.
研究了糜蛋白酶（ChTRP）-Eudragit®（Eu）不溶性复合物形成的条件，旨在将此信息应用于从牛胰脏匀浆的粗滤物中分离糜蛋白酶。ChTRP-Eudragit® L100 的最佳复合物沉淀 pH 值为 4.60，ChTRP-Eudragit® S100 的最佳复合物沉淀 pH 值为 5.40。对于商业酶沉淀，所需的聚电解质浓度低于 0.1%（w/v）。两种聚电解质都抑制了 NaCl 对复合物形成的抑制作用。该方法应用于从牛匀浆中回收酶；通过添加 Eudragit®沉淀糜蛋白酶。通过简单的离心分离不溶性复合物，然后通过将 pH 值改变至 8.20 重新溶解。回收 ChTRP 的最佳条件可获得约 4 倍的纯化因子和 90%的产率。
