Collagenase improved purification from liquid culture from Clostridium histolyticum

摘要

A method for purifying proteins collagenase type I and type II from Clostridium histolyticum from complex unamezcla, comprising the following steps: (a) preparing the complex mixture dissolved in an aqueous liquid phase; (b) forming a precipitate protein collagenase type I and type II I am dissolving the ammonium sulfate in the faselíquida of step (a); (c) separating the precipitate from step (b) of the liquid phase; (d) dissolving the precipitate of step (c ) in an aqueous buffer containing Ca2 + ions 10 and has a pH between 6.0 and 8.0, the conductivity of the buffer yajustar with dissolved to a value between 50 and 300 mS / cm precipitate, forming a complex solution buffered contains proteins collagenase type I and type II (e) extracting the complex buffered solution of step (d) by contacting it with a stationary phase hidrófobay adsorbing proteins collagenase type I and type II on the stationary phase ; (f) stopping the hydrophobic stationary phase along with collagenase proteins adsorbed type I and type II delpaso (e) of the extracted solution; (g) eluting proteins collagenase type I and type II of the stationary phase in step ( f) thereby purifying proteins collagenase type I and type II.