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

D. SPELZINI; G. PICO; B, FARRUGGIA

PROCESS BIOCHEMISTRY

ELSEVIER

Lugar: Amsterdam; Año: 2011 vol. 46 p. 801 - 805

0032-9592

Chymotrypsin was purified from an activated homogenate of bovine pancreas by adsorption onto nonsoluble alginate beads in 25mMTris–acetate–5mMCaCl2 buffer at different adsorbate–adsorbent ratios and the pH values were assaye . Under all the experimental conditions, the enzyme has a positive net electrical charge whereas alginate is negatively charged. After performing steps of washing and desorption  in 25mM Tris–acetate–500mM CaCl2 buffer pH 7.0, chymotrypsin was purified 9 times with an  enzyme recovery of 62%. The eluate fractions resulted in two bands in sodium dodecyl sulphate polyacrylamide  gel electrophoresis (SDS-PAGE). The method allows purification with suitable values from a  raw sample like pancreas homogenate without a previous clarification step2 buffer at different adsorbate–adsorbent ratios and the pH values were assaye . Under all the experimental conditions, the enzyme has a positive net electrical charge whereas alginate is negatively charged. After performing steps of washing and desorption  in 25mM Tris–acetate–500mM CaCl2 buffer pH 7.0, chymotrypsin was purified 9 times with an  enzyme recovery of 62%. The eluate fractions resulted in two bands in sodium dodecyl sulphate polyacrylamide  gel electrophoresis (SDS-PAGE). The method allows purification with suitable values from a  raw sample like pancreas homogenate without a previous clarification step