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. 1973 Jul;133(3):573–584. doi: 10.1042/bj1330573

Covalent chromatography. Preparation of fully active papain from dried papaya latex

Keith Brocklehurst 1, Jan Carlsson 1,*, Marek P J Kierstan 1, Eric M Crook 1
PMCID: PMC1177736  PMID: 4733241

Abstract

1. A Sepharose–(glutathione–2-pyridyl disulphide) conjugate has been prepared. 2. Its use in a new type of chromatography, covalent chromatography by thiol–disulphide interchange, is described. 3. With this technique, papain containing 1 intact catalytic site [thiol with high reactivity towards 2,2′-dipyridyl disulphide (2-Py-S-S-2-Py) at pH4] per mol of protein is readily prepared both from dried papaya latex and from commercial 2×crystallized partially active papain. 4. The catalysis of the hydrolysis of α-N-benzoyl-l-arginine ethyl ester at pH6.0, 25.0°C, I=0.3 by fully active papain thus prepared is characterized by Km=18.2±<0.1mm and kcat.=16.4±0.5s−1.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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