title: A simple method of chymotrypsin concentration and purification from pancreas homogenate using Eudragit® L100 and Eudragit® S100.
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.
authors.affiliation: Physical-Chemistry Department, Faculty of Biochemical and Pharmaceutical Sciences, National University of Rosario, FonCyT and CONICET, Suipacha 570, (S2002RLK) Rosario, Argentina.
authors.forename: Laura Verónica
authors.initials: LV
authors.lastname: Cappella
authors.forename: Valeria
authors.initials: V
authors.lastname: Boeris
authors.forename: Guillermo
authors.initials: G
authors.lastname: Picó
mesh_terms: D000818:Animals; D002417:Cattle; D002498:Centrifugation / Q000379:methods; D002918:Chymotrypsin / Q000302:isolation & purification*; D004591:Electrophoresis, Polyacrylamide Gel; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D009391:Nephelometry and Turbidimetry; D010179:Pancreas / Q000737:chemistry*; D011109:Polymethacrylic Acids / Q000737:chemistry*; D012965:Sodium Chloride / Q000737:chemistry; D012995:Solubility
chemical_list: D011109:Polymethacrylic Acids; C038300:methylmethacrylate-methacrylic acid copolymer; D012965:Sodium Chloride; D002918:Chymotrypsin