The separation of chymotrypsin from a crude filtrate of fresh bovine pancreas homogenate was carried out using precipitation with a commercially available negatively charged natural strong polyelectrolyte: iota-carrageenan. The zymogen form of the enzyme was activated by addition of trypsin (0.0001%, w/w), then, the enzyme was precipitated by polyelectrolyte addition at pH 4.50. The non-soluble complex was separated by simple centrifugation and re-dissolved by a pH change to 8.20. The recovery of chymotrypsin biological activity was 60% of the initial activity in the homogenate with 3-fold increase in its specific activity. The volume of the final product decreased to 10% of the feedstock, concentrating the sample up to 10 times. (C) 2012 Published by Elsevier Ltd.