A simple method of chymotrypsin concentration and purification from pancreas homogenate using Eudragit^® L100 and Eudragit^® S100

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• 作者：Laura Veró ; nica Cappella^a ; Valeria Boeris ; ^a ; ^{valeriaboeris@conicet.gov.ar} ; ^{valeriaboeris@hotmail.com} ; Guillermo Picó ; ^a
• 关键词：Polyelectrolyte precipitation ; Chymotrypsin ; Eudragit ; Protein purification
• 刊名：Journal of Chromatography B, Analytical Technologies in the Biomedical and Life Sciences
• 出版年：2011
• 出版时间：15 April 2011
• 年：2011
• 卷：879
• 期：13-14
• 页码：1003-1007
• 全文大小：420 K

文摘

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.