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A simple method of chymotrypsin concentration and purification from pancreas homogenate using Eudragit® L100 and
Laura Verónica Cappella1, Valeria Boeris, Guillermo Picó
1Physical-Chemistry Department, Faculty of Biochemical and Pharmaceutical Sciences, National University of Rosario, FonCyT and CONICET, Suipacha 570, (S2002RLK) Rosario, Argentina.
Abstract:
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.
