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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Marston, F., Lowe, P., Doel, M. et al. Purification of Calf Prochymosin (Prorennin) Synthesized in Escherichia Coli. Nat Biotechnol 2, 800–804 (1984). https://doi.org/10.1038/nbt0984-800
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DOI: https://doi.org/10.1038/nbt0984-800
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