Methods for improving multiple protein properties

Abstract

The present invention provides efficient methods for obtaining a protein with one or more beneficial attributes in industrial, consumer or pharmaceutical applications. In some preferred embodiments, the present invention provides methods for producing superior enzymes for a given application through screening an abbreviated set of candidate enzymes.

1 - 34 . (canceled) 
     
     
         35 . A method for producing a substrate stain-specific enzyme variant, comprising:
 a) determining the zeta potential of a substrate stain in a reference buffer;   b) determining the zeta potential of a parent enzyme in the reference buffer; and   c) producing a substrate stain-specific enzyme variant by introducing at least one amino acid modification in said parent enzyme, such that said zeta potential of said substrate stain-specific enzyme variant is closer to the zeta potential of said substrate stain than to said zeta potential of said parent enzyme.   
     
     
         36 . The method of  claim 35 , wherein said modification comprises at least one amino acid substitution, deletion and/or insertion. 
     
     
         37 . The method of  claim 35 , wherein said modification comprises chemical modification of said parent enzyme. 
     
     
         38 . The method of  claim 35 , wherein said substrate stain-specific enzyme variant is positively charged and endothermic, and said substrate stain is negatively charged. 
     
     
         39 . The composition of  claim 35 , wherein said stain-specific enzyme variant is negatively charged and exothermic, and said substrate stain is negatively charged. 
     
     
         40 . The composition of  claim 35 , wherein said stain-specific enzyme variant is positively charged and exothermic, and the substrate stain is positively charged. 
     
     
         41 . The composition of  claim 35 , wherein said stain-specific enzyme variant is negatively charged and endothermic, and said substrate stain is positively charged. 
     
     
         42 . A method for producing a composition for cleaning multiple stains comprising:
 a) determining the zeta potential of each of said multiple stains in a reference buffer;   b) selecting a cleaning enzyme having a zeta potential in said reference buffer that is essentially equal to the zeta potential of at least one of said multiple stains; and   c) producing a composition for cleaning multiple stains, wherein said composition comprises at least one cleaning enzyme selected in step b).   
     
     
         43 . The method of  claim 42 , wherein said composition comprises a detergent solution with a pH and conductivity that is essentially equal to that of said reference buffer. 
     
     
         44 . The method of  claim 42 , wherein said selecting step identifies more than one cleaning enzyme. 
     
     
         45 . The method of  claim 42 , wherein said composition comprises at least two cleaning enzymes and wherein said at least two cleaning enzymes have zeta potentials that correspond to said zeta potentials of at least two of said multiple stains. 
     
     
         46 . The composition for cleaning multiple stains produced using the method of  claim 42 . 
     
     
         47 . The composition of  claim 46 , wherein said composition comprises at least one cleaning enzyme. 
     
     
         48 . The composition of  claim 47 , wherein said at least one cleaning enzyme is a variant protein. 
     
     
         49 . The composition of  claim 48 , wherein said variant protein is more negatively charged than the wild-type precursor protein used to produce said variant protein. 
     
     
         50 . The composition of  claim 46 , wherein said variant protein is more positively charged than the wild-type precursor protein used to produce said variant protein. 
     
     
         51 . The composition of  claim 46 , wherein said variant protein is more negatively charged than the wild-type precursor protein used to produce said variant protein for enhanced stability in detergents containing anionic surfactants.