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  • Title: Studies on the isolation of penicillin acylase from Escherichia coli by aqueous two-phase partitioning.
    Author: Guan Y, Wu XY, Treffry TE, Lilley TH.
    Journal: Biotechnol Bioeng; 1992 Aug 05; 40(4):517-24. PubMed ID: 18601146.
    Abstract:
    A method of enzyme release and aqueous two-phase extraction is described for the separation of penicillin acylase from Escherichia coli cells. Butyl acetate, 12% (v/v), treatment combined with freeze-thawing gives up to 70% enzyme release. For polyethylene glycol (PEG) + phosphate two-phase extraction systems the enzyme purity and yield were rather low. Modified PEG, including PEG-ampicillin, PEG-aniline, PEG-phosphate, and PEG-trimethylamine, were synthesized and used in aqueous two-phase systems; PEG-trimethylamine is the most satisfactory. A system containing 12% (w/w) PEG4000, 8% (w/w) of which is PEG-trimethylamine, with 0.7M potassium phosphate at pH 7.2, resulted in the enzyme selective partition being greatly enhanced by charge directed effects. Possible mechanisms for the separation process are discussed.
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