INVESTIGADORES
TUBIO Gisela
artículos
Título:
Extraction Of Trypsin From Bovine Pancreas By Applying Polyethyleneglycol/Sodium Citrate Aqueous Two-Phase Systems
Autor/es:
TUBIO, GISELA;; PICO, GUILLERMO; NERLI, BIBIANA
Revista:
Journal of Chromatography B
Editorial:
ELSEVIER SCIENCE BV
Referencias:
Lugar: Amsterdam; Año: 2009 vol. 877 p. 115 - 120
ISSN:
1570-0232
Resumen:
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 polyethylenglycol/sodium citrate (PEG/NaCit) aqueous two-phase systems. Partitioning behaviour of TRP and ChTRP 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 3,350 and NaCit pH 5.20 showed the best separation capability. Addition of NaCl up to a final concentration of 7 % (wt/wt) and decreasing top/bottom volume ratio to 0.1 led to recover the 60 % of pancreatic TRP in a concentrated form in the top phase with a 3-fold purification. Biomass presence up to 25 % (wt/wt) of total system mass did not affect significantly yield and purification parameters.