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Isolation and purification of plastocyanin from spinach stored frozen using hydrophobic interaction and ion-exchange chromatography

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Abstract

A new simple three-day procedure for preparative isolation and purification of plastocyanin from spinach stored in the frozen state is described. This procedure is based on batch adsorption on ion-exchange resin, ammonium sulphate precipitation, and purification on a Phenyl-Sepharose hydrophobic interaction column and a single Q Sepharose High Performance ion-exchange column. Approximately 100 mg of plastocyanin with an absorbance ratio A278/A597 of 1.10±0.02 in the oxidized state was typically obtained from 12 kg of spinach leaves. The purified spinach plastocyanin is shown to be homogeneous to the resolution of free solution capillary electrophoresis.

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Abbreviations

MES:

2(N-morpholino)ethanesulfonic acid

Tris:

Tris(hydroxymethyl)aminomethane

FSCE:

free solution capillary electrophoresis

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Authors and Affiliations

  1. Chemistry Department A, The Technical University of Denmark, Building 207, DK-2800, Lyngby, Denmark

    Hans E. M. Christensen, Lars S. Conrad & Jens Ulstrup

Authors
  1. Hans E. M. Christensen
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  2. Lars S. Conrad
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  3. Jens Ulstrup
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Christensen, H.E.M., Conrad, L.S. & Ulstrup, J. Isolation and purification of plastocyanin from spinach stored frozen using hydrophobic interaction and ion-exchange chromatography. Photosynth Res 28, 89–93 (1991). https://doi.org/10.1007/BF00033718

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  • DOI: https://doi.org/10.1007/BF00033718

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