Folding and activation of human procathepsin S from inclusion bodies produced in Escherichia coli.

@article{Kopitar1996FoldingAA,
  title={Folding and activation of human procathepsin S from inclusion bodies produced in Escherichia coli.},
  author={Gregor Kopitar and Marko Dolinar and B. Strukelj and J. Pungercar and Vito Turk},
  journal={European journal of biochemistry},
  year={1996},
  volume={236 2},
  pages={
          558-62
        }
}
Human procathepsin S was produced in the form of insoluble inclusion bodies in Escherichia coli using an inducible T7-based expression system. After cell disruption, the dissolved inclusion body proteins were S-sulphonated with 2-nitro-5-thiosulphobenzoate and purified by gel filtration. Recombinant procathepsin S was renatured at pH 7.6 by a two-step dilution which significantly increased the yield of production compared to single-step dilution. The proenzyme was autocatalytically processed to… 
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Refolding is the most critical step in the process of generating active cysteine proteases and the current approaches to refolding include dialysis, dilution and chromatography, while purification is mainly achieved by various column chromatography.
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