Extraction of trypsin from bovine pancreas by applying polyethyleneglycol/sodium citrate aqueous two-phase systems

被引：32

作者：

Tubio, Gisela ^{[1
]}

Pico, Guillermo A. ^{[1
]}

Nerli, Bibiana B. ^{[1
]}

机构：

[1] Natl Univ Rosario, Fac Biochem & Pharmaceut Sci, FonCyT, Dept Chem Phys,Bioseparat Lab,CONICET, Rosario, Santa Fe, Argentina

来源：

JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES | 2009年 / 877卷 / 03期

关键词：

Trypsin; Chymotrypsin; Partitioning; Purification; Aqueous two-phase systems; Pancreatic proteases; SODIUM-CITRATE; PENICILLIN ACYLASE; MOLECULAR-WEIGHT; PURIFICATION; CHYMOTRYPSIN; PARTITION; ELASTASE; FEATURES; MILK;

D O I：

10.1016/j.jchromb.2008.11.016

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

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. (c) 2008 Elsevier B.V. All rights reserved.

引用

页码：115 / 120

页数：6

共 21 条

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