Abstract
This article studies the influence of pH and extraction solution, time and solid/liquid ratio on the extraction of protease from ora-pro-nobis. Aqueous two-phase systems (ATPS) formed by polyethylene glycol 1500 or 4000 and phosphate was investigated as a purification step for these proteases. The best results were with sodium phosphate pH 7.0, 60 min of extraction, and at the liquid/solid ratio 21 mL/g. ATPS formed by polyethylene glycol and phosphate were obtained. The increase in molar mass resulted in the expansion of the biphasic region, exclusion volume, and the free volume of the upper phase. The proteases were preferentially partitioned to the upper phase. The free energy indicates that the partition of proteases to the upper phase is a spontaneous process. High selectivity, activity recovery, and the purification factor indicate that the studied systems can be used as a good alternative for purification of this enzyme.
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