A Modified Procedure for the Purification of Clostridial Collagenase
- 1 January 1978
- journal article
- research article
- Published by Taylor & Francis in Preparative Biochemistry
- Vol. 8 (5) , 387-407
- https://doi.org/10.1080/10826067809412290
Abstract
A method is described for the purification of clostridial [Clostridium histolyticum] collagenase from a crude enzyme preparation employing cation exchange chromatography on SP Sephadex, anion exchange chromatography on DEAE cellulose and gel filtration on Sephacryl S-2000. Emphasis was placed on purity using continuous shallow gradients for the ion exchange separations to increase resolution and monitoring eluates both with respect to UV light absorption at 230 nm and analytical disc gel acrylamide electrophoresis. Protein tractions were assayed for collagenolytic and non-specific proteolytic activity. The purity of the final preparation was assessed by acrylamide electrophoresis, gel filtration and amino acid analysis. The isolated enzyme hydrolyzed between 30 and 40% of rat tail tendon collagen in 1 h at 37.degree. C and lacked measurable trypsin or elastase-like activity.This publication has 14 references indexed in Scilit:
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