Purification of acetohydroxy acid synthase by separation in an aqueous two-phase system

S. Engel; Z. Barak; D. M. Chipman; J. C. Merchuk

doi:10.1016/S0378-4347(00)00053-0

Purification of acetohydroxy acid synthase by separation in an aqueous two-phase system

S. Engel, Z. Barak, D. M. Chipman, J. C. Merchuk

Research output: Contribution to journal › Article › peer-review

8 Scopus citations

Abstract

Extraction in a polyethylene glycol (PEG)-phosphate aqueous two-phase system was considered as a primary step in purification of the acetohydroxy acid synthase III large catalytic subunit from an E. coli extract. Extraction optimization was achieved by varying the system parameters. Two systems with the following weight compositions were chosen for purification: PEG-2000 (16%)-phosphate (6%) and PEG-4000 (14%)-phosphate (5.5%)-KCl (8%), both at pH 7.0 and 1 mg total protein per 1 g system. Significant purification was achieved by a single extraction step with 70% recovery of the enzyme. After an additional ion-exchange chromatography step, pure enzyme was obtained in a 50% overall yield. (C) 2000 Elsevier Science B.V.

Original language	English
Pages (from-to)	281-286
Number of pages	6
Journal	Journal of Chromatography B: Biomedical Sciences and Applications
Volume	743
Issue number	1-2
DOIs	https://doi.org/10.1016/S0378-4347(00)00053-0
State	Published - 23 Jun 2000

Keywords

Acetohydroxy acid synthase
Aqueous two-phase systems
Enzymes
Partitioning

ASJC Scopus subject areas

General Chemistry

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10.1016/S0378-4347(00)00053-0

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abstract = "Extraction in a polyethylene glycol (PEG)-phosphate aqueous two-phase system was considered as a primary step in purification of the acetohydroxy acid synthase III large catalytic subunit from an E. coli extract. Extraction optimization was achieved by varying the system parameters. Two systems with the following weight compositions were chosen for purification: PEG-2000 (16%)-phosphate (6%) and PEG-4000 (14%)-phosphate (5.5%)-KCl (8%), both at pH 7.0 and 1 mg total protein per 1 g system. Significant purification was achieved by a single extraction step with 70% recovery of the enzyme. After an additional ion-exchange chromatography step, pure enzyme was obtained in a 50% overall yield. (C) 2000 Elsevier Science B.V.",

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AU - Barak, Z.

AU - Chipman, D. M.

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KW - Acetohydroxy acid synthase

KW - Aqueous two-phase systems

KW - Enzymes

KW - Partitioning

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Purification of acetohydroxy acid synthase by separation in an aqueous two-phase system

Abstract

Keywords

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