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

L. CAPELLA; V. BOERIS; G. PICO

Journal of Chromatography B

ELSEVIER SCIENCE BV

Lugar: Amsterdam; Año: 2011 vol. 879 p. 1003 - 1007

1570-0232

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 pHchange to 8.20. The best conditions to recover ChTRP brought about a purification   factor of around 4 and 90% yield.® (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 pHchange to 8.20. The best conditions to recover ChTRP brought about a purification   factor of around 4 and 90% yield.