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Extraction of Trypsin From Bovine Pancreas by Applying polyethyleneglycol/sodium Citrate Aqueous Two-Phase Systems

Extraction of Trypsin From Bovine Pancreas by Applying polyethyleneglycol/sodium Citrate Aqueous Two-Phase Systems

Gisela Tubio, Guillermo A Picó, Bibiana B Nerli

J Chromatogr B Analyt Technol Biomed Life Sci. 2009 Jan 15;877(3):115-20.

PMID: 19109075

Abstract:

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.

Chemicals Related in the Paper:

Catalog Number	Product Name	CAS Number	Price
AP9001927-B	Protease from bovine pancreas	9001-92-7	Price
AP9002077	Trypsin from bovine pancreas	9002-07-7	Price
AP9004073	α-Chymotrypsin from bovine pancreas	9004-07-3	Price

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