Enzyme multimer and process of producing same

Abstract

An enzyme multimer is provided which comprises a plurality of monomer units including a first monomer unit having enzymatic activity and a second monomer unit having enzymatic activity, wherein each of the first monomer unit and the second monomer unit comprise a cysteine residue and the cysteine residues of each monomer unit are covalently bound to each other through a disulfide bond to covalently attach the first monomer unit to the second monomer unit.

1 . An enzyme multimer comprising a plurality of monomer units including a first monomer unit having enzymatic activity and a second monomer unit having enzymatic activity, wherein each of said first monomer unit and said second monomer unit comprise a cysteine residue and said cysteine residues are covalently bound to each other through a disulfide bond to covalently attach said first monomer unit to said second monomer unit.  
     
     
         2 . The enzyme multimer according to  claim 1 , wherein said first and second monomer units in said enzyme multimer are derived from a precursor enzyme which has enzymatic activity.  
     
     
         3 . The enzyme multimer according to  claim 2 , wherein said first or second monomer unit differs from its corresponding precursor enzyme by the substitution or addition of a cysteine residue therein.  
     
     
         4 . The enzyme multimer according to  claim 2 , wherein said precursor enzyme comprises a hydrolase, an oxidoreductase, a transferase, a lyase or a ligase.  
     
     
         5 . The enzyme multimer according to  claim 4 , wherein said first or second monomer comprises protease, lipase, amylase, β-galactosidase, cellulase, lactase, polygalacturonase, β-glucoamylase, esterase or hemicellulase.  
     
     
         6 . The enzyme multimer according to  claim 4 , wherein said first or second monomer comprises peroxidase, oxidase, laccase, glucose oxidase or ligninase.  
     
     
         7 . The enzyme multimer according to  claim 4 , wherein said first or second monomer comprises NADH reductase or 2,5DKG reductase.  
     
     
         8 . The enzyme multimer according to  claim 1 , wherein said first or second monomer comprises a protease derived from  Bacillus.    
     
     
         9 . The enzyme multimer according to  claim 1 , wherein said  Bacillus  comprises  Bacillus lentus, Bacillus subtilis, Bacillus amyloliquefaciens; Bacillus stearothermophilus  or  Bacillus licheniformis.    
     
     
         10 . The enzyme multimer according to  claim 3 , wherein said precursor enzyme comprises a protease derived from  Bacillus.    
     
     
         11 . The enzyme multimer according to  claim 10 , wherein said  Bacillus  comprises  Bacillus lentus, Bacillus subtilis, Bacillus amyloliquefaciens; Bacillus stearothermophilus  or  Bacillus licheniformis.    
     
     
         12 . A method for producing an enzyme multimer comprising: 
 (a) preparing a mutant enzyme through an addition of or substitution with a cysteine residue in a precursor enzyme;    (b) preparing a second enzyme comprising a cysteine residue; and    (c) mixing the mutant enzyme of step (a) with said second enzyme of step (b) under conditions so as to form a disulfide bond between said added or substituted cysteine residue of said mutant enzyme and said cysteine residue in said second enzyme.    
     
     
         13 . The composition according to  claim 11 , wherein said monomer comprises a protease, a cellulase, an amylase, a lipase, an esterase, a hemicellulase, an oxido-reductase, hydrolase, anoxid-redactase, a transferase, a lyase or a ligase.  
     
     
         14 . The composition according to  claim 12 , wherein said monomer comprises a protease.  
     
     
         15 . The composition according to  claim 13 , wherein said monomer comprises a protease derived from  Bacillus.    
     
     
         16 . The composition according to  claim 14 , wherein said  Bacillus  comprises  Bacillus lentus, Bacillus subtilis, Bacillus amyloliquefaciens; Bacillus stearothermophilus  or  Bacillus licheniformis.    
     
     
         17 . A detergent composition comprising the multimeric enzyme of  claim 1 .  
     
     
         18 . A detergent composition according to  claim 16 , wherein said multimeric enzyme comprises a protease, an amylase, a cellulase, a lipase, a xylanase or a mixture thereof.  
     
     
         19 . A method for treating textiles comprising contacting a suitable textile composition with a multimeric enzyme according to  claim 1  comprising α-amylase, cellulase, protease or esterase.  
     
     
         20 . A method for liquefying starch comprising contacting a multimeric enzyme composition according to  claim 1  with a starch solution, wherein said multimeric enzyme comprises α-amylase, cellulase, esterase or hemicellulase.  
     
     
         21 . A method for treating pulp and paper comprising contacting a multimeric enzyme composition according to  claim 1  with a pulp mixture, wherein said multimeric enzyme comprises cellulase, hemicellulase, lipase or esterase.