info:eu-repo/semantics/article
Purification of chymotrypsin from pancreas homogenate by adsorption onto non-soluble alginate beads
Fecha
2011-03Registro en:
Spelzini, Darío; Farruggia, Beatriz Monica; Picó, Guillermo Alfredo; Purification of chymotrypsin from pancreas homogenate by adsorption onto non-soluble alginate beads; Elsevier; Process Biochemistry; 46; 3; 3-2011; 801-805
0032-9592
1359-5113
CONICET Digital
CONICET
Autor
Spelzini, Darío
Farruggia, Beatriz Monica
Picó, Guillermo Alfredo
Resumen
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.