Design and optimization of an alternative chymotrypsin purification method by adsorption onto non-soluble alginate–carrageenan bed

BÁRBARA, BOSIO; EMILIA, BRASSESCO MARÍA; GASTÓN, KNOBEL; GUILLERMO, PICÓ; NADIA, WOITOVICH VALETTI

POLYMER BULLETIN

SPRINGER

Lugar: Berlin; Año: 2020

0170-0839

chymotrypsinusing adsorption onto non-soluble alginate?carrageenan bed. In the firstinstance, an experimental statistical design was used to screen which parameters aresignificant in the adsorption process. The variables tested were pH, temperature, andmedium?s ionic strength. Significant statistical differences (p < 0.05) were obtainedfor pH and the medium?s ionic strength, whereas temperature had no influence onthe adsorption efficiency in the studied range. In a second step, response surfacemethodology was used to find the conditions that produced the highest adsorptionof chymotrypsin. The optimal working conditions found were pH 6.5 without theaddition of salts, which is consistent with an electrostatic interaction. The adsorptionkinetics were studied, showing that the adsorption of the enzyme followed a pseudofirst-order rate expression. Different adsorption isotherm models were applied todescribe the equilibrium isotherms. The Freundlich model was found to be moresuitable for our data. At optimal conditions, 1 g of hydrated matrix adsorbed 11 mgof chymotrypsin. In the desorption process, 93.1% of the biological activity of chymotrypsinwas recovered in only 20 min. Additionally, experimental breakthroughprofiles were acquired. The optimized conditions were applied in the design of bothbatch and packed bed adsorption systems for the purification of chymotrypsin fromfresh bovine pancreas, reaching a purification factor of 3.8 and 6.9, respectively.