Purification of chymotrypsin from pancreas homogenate by adsorption onto non-soluble alginate beads

Abstract

Chymotrypsin was purified from an activated homogenate of bovine pancreas by adsorption onto non-soluble alginate beads in 25 mM Tris-acetate-5 mM CaCl2 buffer at different adsorbate-adsorbent ratios and the pH values were assayed. 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 25 mM Tris-acetate-500 mM CaCl2 buffer ρH 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.