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Purification of Calf Prochymosin (Prorennin) Synthesized in Escherichia Coli

Bio/Technology volume 2, pages 800804 (1984) | Download Citation

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Abstract

Recombinant calf prochymosin synthesized in E. coli was shown to accumulate in the form of insoluble inclusion bodies. Isolation of this aggregated material, combined with specific washing procedures, was the most significant stage of the purification protocol. Disruption of proteins in the inclusion bodies necessitated denaturation and renaturation. The method described can completely solubilize prochymosin. At this stage acidification produced active chymosin. Subsequently, ion-exchange chromatography produced highly purified prochymosin which, after acidification, yielded chymosin >99% pure.

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Affiliations

  1. Department of Fermentation Development and Downstream Processing, Celltech Ltd., 250 Bath Road, Slough SL1 4DY, Berkshire, U.K.

    • Fiona A. O. Marston
    • , Peter A. Lowe
    • , Susan White
    •  & Sarojani Angal
  2. Department of Molecular Biology, Celltech Ltd., 250 Bath Road, Slough SL1 4DY, Berkshire, U.K.

    • Michael T. Doel
  3. Present address: Department of Microbiology, The University of Chicago, Cummings Life Science Center, 920 East 58th Street, Chicago, Illinois 60637, U.S.A.

    • Joyce M. Schoemaker

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DOI

https://doi.org/10.1038/nbt0984-800

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