Extraction of trypsin from bovine pancreas by applying Polyethyleneglycol/sodium citrate aqueous two-phase Systems.

TUBÍO, GISELA; PICÓ, GUILLERMO; NERLI, BIBIANA

Journal of Chromatography B

Elsevier Science Ltd.

Año: 2009 vol. 877 p. 115 - 120

1570-0232

The goal of this work was to determine the optimal conditions for separating trypsin (TRP) from alphachymotrypsin (ChTRP) and to apply them for trypsin purification from bovine pancreas by liquid?liquid extraction with polyethyleneglycol/sodium citrate (PEG/NaCit) aqueous two-phase systems. Partitioning behaviours of TRP and ChTRP are demonstrated to be very sensitive to variables such as PEG molecular weight, pH and tie line length. Aqueous two-phase systems (ATPSs) formed by PEG ofMW3350 and NaCit pH 5.20 showed the best separation capability. The addition of NaCl up to a final concentration of 7% (w/w) and the decrease of top/bottom volume ratio to 0.1 led to the recovery of 60% of pancreatic TRP in a concentrated form in the top phase with a 3-fold purification. Biomass presence up to 25% (w/w) of the total system mass did not affect significantly yield and purification parameters.