Journal
JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES
Volume 807, Issue 1, Pages 69-74Publisher
ELSEVIER
DOI: 10.1016/j.jchromb.2004.03.030
Keywords
aqueous two-phase systems; Methanococcus jannaschii; partitioning; thermoseparation; alpha-amylase; amyloglucosidase
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Purification of a recombinant, thermostable alpha-amylase (MJA1) from the hyperthermophile, Methanococcus jannaschii, was investigated in the ethylene oxide-propylene oxide random copolymer (PEO-PPO)/(NH4)(2)SO4, and poly(ethylene glycol) (PEG)/(NH4)(2)SO4 aqueous two-phase systems. MJA1 partitioned in the top polymer-rich phase, while the remainder of proteins partitioned in the bottom salt-rich phase. It was found that enzyme recovery of up to 90% with a purification factor of 3.31 was achieved using a single aqueous two-phase extraction step. In addition, the partition behavior of pure amyloglucosidase in polymer/salt aqueous two-phase systems was also evaluated. All of the studied enzymes partitioned unevenly in these polymer/salt systems. This work is the first reported application of thermoseparating polymer aqueous two-phase systems for the purification of extremophile enzymes. (C) 2004 Published by Elsevier B.V.
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