Extraction of trypsin from bovine pancreas by applying polyethyleneglycol/sodium citrate aqueous two-phase systems.
The goal of this work was to determine the optimal conditions for separating trypsin (TRP) from alpha-chymotrypsin (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 of MW 3350 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.
Tubio
Gisela
G
Chemical Physics Department, CONICET, FonCyT, Faculty of Biochemical and Pharmaceutical Sciences, National University of Rosario, Rosario, Argentina.
Picó
Guillermo A
GA
Nerli
Bibiana B
BB
D000818:Animals; D018533:Biomass; D002417:Cattle; D005591:Chemical Fractionation / Q000379:methods*; D002918:Chymotrypsin / Q000737:chemistry / Q000302:isolation & purification; D002951:Citrates / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008970:Molecular Weight; D010179:Pancreas / Q000737:chemistry*; D011092:Polyethylene Glycols / Q000737:chemistry*; D012965:Sodium Chloride / Q000737:chemistry; D000077559:Sodium Citrate; D014357:Trypsin / Q000737:chemistry / Q000302:isolation & purification*
D002951:Citrates; D000077559:Sodium Citrate; D011092:Polyethylene Glycols; D012965:Sodium Chloride; D002918:Chymotrypsin; C117608:alpha-chymotrypsin; D014357:Trypsin