Blends containing proteases

Abstract

Described are compositions, in particular lyophilizates, containing proteolytic enzymes, and methods for producing the compositions. Typically these compositions contain one or more proteases with collagenase activity and a neutral protease, for example, thermolysin. The compositions are free of acetate salts. Surprisingly, such compositions can be dissolved in water more rapidly than lyophilized protease mixtures of the state of the art.

What is claimed is: 
     
         1 . A method for producing a solid composition comprising the steps of:
 preparing a homogeneous solution of an acetate-free preparation of a neutral protease (NP) in an aqueous acetate-free, low-salt solution, wherein the low-salt solution has an aggregate concentration of salt(s) in the range of about 1 mM to about 250 mM;   adding a neutral salt to the homogeneous solution and dissolving the neutral salt, thereby making a stabilized solution, wherein the stabilized solution additionally comprises a buffer salt buffering in a range of about pH 6 to about pH 8.5, and wherein the stabilized solution further comprises calcium chloride;   mixing the stabilized solution with an acetate-free preparation of one or more proteolytic enzymes with collagenase activity (C) and making a homogeneous solution; and   freeze-drying the homogeneous solution, thereby obtaining the solid composition.   
     
     
         2 . The method according to  claim 1 , wherein the neutral protease is thermolysin from  Bacillus thermoproteolyticus.    
     
     
         3 . The method according to  claim 1 , wherein a weight/weight ratio of the neutral protease versus all proteases present in the composition (NP/(NP+C)) is in a range of about 1 to about 25. 
     
     
         4 . The method according to  claim 1 , wherein the neutral salt is sodium chloride and a weight/weight ratio of all proteases present in the composition versus sodium chloride ((NP+C)/NaCl) is in a range of about 0.1 to about 5. 
     
     
         5 . The method according to  claim 1 , wherein a weight/weight ratio of all proteases present in the composition versus calcium chloride ((NP+C)/CaCl 2 ) is in a range of about 10 to about 500. 
     
     
         6 . The method according to  claim 1 , wherein a weight/weight ratio of all proteases present in the composition versus the buffer salt ((NP+C)/buffer) is in a range of about 0.05 to about 2. 
     
     
         7 . The method of  claim 1 , wherein the solid composition further comprises sodium chloride, calcium chloride, and an organic buffer salt. 
     
     
         8 . The method of  claim 1  further comprising the step of contacting the solid composition obtained in said step of freeze-drying with water. 
     
     
         9 . The method of  claim 8 , wherein upon said further step of contacting, the proteases in solution have a concentration in a range of about 5 mg/ml to about 50 mg/ml. 
     
     
         10 . The method of  claim 9 , wherein upon said further step of contacting, a homogeneous solution is obtained within a period of about 3 minutes or less than 3 minutes.