Combinatorial enzyme development

Abstract

Disclosed is a process for obtaining an enzyme having a specified enzyme activity derived from a heterogeneous DNA population by screening, for the specified enzyme activity, a library of clones containing DNA from the heterogeneous DNA population which have been exposed to directed mutagenesis towards production of the specified enzyme activity. Also disclosed is a process for obtaining an enzyme having a specified enzyme activity by screening, for the specified enzyme activity, a library of clones containing DNA from a pool of DNA populations which have been exposed to directed mutagenesis in an attempt to produce in the library of clones DNA encoding an enzyme having one or more desired characteristics which can be the same or different from the specified enzyme activity.

What is claimed is:  
     
         1 . A process for obtaining an enzyme having a specified enzyme activity derived from a heterogeneous DNA population, which process comprises: 
 screening, for the specified enzyme activity, a library of clones containing DNA from the heterogeneous DNA population which have been exposed to directed mutagenesis towards production of the specified enzyme activity.    
     
     
         2 . The process of  claim 1  which further comprises, prior to said directed mutagenesis, selectively recovering from the heterogeneous DNA population DNA which comprises DNA sequences coding for a common characteristic, which can be the same or different from the specified enzyme activity.  
     
     
         3 . The process of  claim 2  wherein recovering the DNA preparation comprises contacting the DNA population with a specific binding partner for at least a portion of the coding sequences.  
     
     
         4 . The process of  claim 3  wherein the specific binding partner is a solid phase bound hybridization probe.  
     
     
         5 . The process of  claim 2  wherein the common characteristic is a class of enzyme activity.  
     
     
         6 . The process of  claim 5  which comprises recovering DNA from clones containing DNA from the heterogeneous DNA population which exhibit the class of enzyme activity.  
     
     
         7 . The process of  claim 1  wherein the directed mutagenesis is site-specific directed mutagenesis.  
     
     
         8 . The process of  claim 1  which further comprises prescreening said library of clones for an activity, which can be the same or different from the specified enzyme activity, prior to exposing them to directed mutagenesis.  
     
     
         9  The process of  claim 8  which comprises prescreening said clones for the expression of a protein of interest.  
     
     
         10 . A process for obtaining an enzyme having a specified enzyme activity, which process comprises: 
 screening, for the specified enzyme activity, a library of clones containing DNA from a pool of DNA populations which have been exposed to directed mutagenesis in an attempt to produce in the library of clones DNA encoding an enzyme having one or more desired characteristics which can be the same or different from the specified enzyme activity.    
     
     
         11 . The process of  claim 10  which further comprises, prior to said directed mutagenesis, selectively recovering from the heterogeneous DNA population DNA which comprises DNA sequences coding for at least one common enzyme characteristic, which can be the same or different from the specified enzyme activity.  
     
     
         12 . The process of  claim 11  wherein the at least one common enzyme characteristic is at least one common enzyme activity.  
     
     
         13 . The process of  claim 12  wherein recovering the DNA preparation comprises contacting the DNA population with a specific binding partner for at least a portion of the coding sequences.  
     
     
         14 . The process of  claim 13  wherein the specific binding partner is a solid phase bound hybridization probe.  
     
     
         15 . The process of  claim 14  which comprises recovering DNA from clones containing DNA from the heterogeneous DNA population which exhibit the class of enzyme activity.  
     
     
         16 . The process of  claim 10  wherein the directed mutagenesis is site-specific directed mutagenesis.