Laccase activity enhancers

Abstract

New mediators for use in laccase-mediator systems (LMS) are disclosed. The mediators enhance the activity of enzymes exhibiting laccase activity and can be used, for example, to bleach a dye in solution, to oxidize an appropriate compound, or to initiate vinyl or phenol polymerizations. A composition comprising an enzyme exhibiting laccase activity and an enzyme enhancing agent (also referred to as a mediator or an activating agent) is also disclosed. A process for oxidizing a substrate that comprises treating the substrate with a composition comprising an enzyme exhibiting laccase activity and an enzyme enhancing agent is also disclosed.

What is claimed is:  
     
         1 . A composition comprising an enzyme exhibiting laccase activity and an enzyme enhancing agent selected from: 
 (i) a monomer, dimer, or oligomer of at least one of:                          wherein:    R 1  is OCH 3 , H, or another moiety;    R 2  is OCH 3 , H, or another moiety;    R 3  is OH, sulfonate, or another moiety;    R 4  is an OH or methylene that connects to another monomer; and    R 5  is SO 3 H or H;    (ii) N-benzylidene benzylamine or a substituted N-benzylidene benzylamine having the structure:                          wherein:    R 6  is H, OCH 3 , NH(CH 3 ) 2 , Cl, Br, an aliphatic group, or an aromatic group; and    R 7  is H, OCH 3 , NH(CH 3 ) 2 , Cl, Br, an aliphatic group, or an aromatic group;    (iii) a derivative of cinnamic acid having the structure:                          wherein:    R 8  is OCH 3 , NH2, Cl, NH(CH 3 ) 2 ; and    R 9  is OH or H;    (vi) salicylic acid;    (v) epigallocatechin gallate;    (vi) melamine; and    (vii) 3,4-dihydroxybenzyl alcohol or a substituted 3, 4-dihydroxybenzyl alcohol.    
     
     
         2 . The composition of  claim 1  wherein said enzyme exhibiting laccase activity is selected from a laccase enzyme of enzyme classification EC 1.10.3.2, a catechol oxidase enzyme of enzyme classification EC 1.10.3.1, a monophenol monooxygenase enzyme of enzyme classification EC 1.14.99.1, a bilirubin oxidase enzyme of enzyme classification EC 1.3.3.5, and an ascorbate oxidase enzyme of enzyme classification EC 1.10.3.3.  
     
     
         3 . The composition of  claim 1  that further comprises a hydrolase.  
     
     
         4 . The composition of  claim 3  wherein said hydrolase is a xylanase.  
     
     
         5 . The composition of  claim 1  wherein said monomer, dimer, or oligomer is an organic soluble fraction of lignin or a lignin sulfonate.  
     
     
         6 . The composition of  claim 1  wherein said derivative of cinnamic acid is selected from: 
 4-(dimethylamino)cinnamic acid; 3-hydroxy-4-methyoxycinnamic acid; 4-aminocinnamic acid; and 4-chlorocinnamic acid.  
 
     
     
         7 . A process for oxidizing a substrate, comprising treating the substrate with a composition comprising an enzyme exhibiting laccase activity and an enzyme enhancing agent selected from: 
 (i) a monomer, dimer, or oligomer of at least one of:                          wherein:    R 1  is OCH 3 , H, or another moiety;    R 2  is OCH 3 , H, or another moiety;    R 3  is OH, sulfonate, or another moiety;    R 4  is an OH or methylene that connects to another monomer; and    R 5  is SO 3 H or H;    (ii) N-benzylidene benzylamine or a substituted N-benzylidene benzylamine having the structure:                          wherein:    R 6  is H, OCH 3 , NH(CH 3 ) 2 , Cl, Br, an aliphatic group, or an aromatic group; and    R 7  is H, OCH 3 , NH(CH 3 ) 2 , Cl, Br, an aliphatic group, or an aromatic group;    (iii) a derivative of cinnamic acid having the structure:                          wherein:    R 8  is OCH 3 , NH 2 , Cl, NH(CH 3 ) 2 ; and    R 9  is OH or H;    (vi) salicylic acid;    (v) epigallocatechin gallate;    (vi) melamine; and    (vii) 3,4-dihydroxybenzyl alcohol or a substituted 3,4-dihydroxybenzyl alcohol.    
     
     
         8 . The process of  claim 7  wherein said enzyme exhibiting laccase activity is selected from a laccase enzyme of enzyme classification EC 1.10.3.2, a catechol oxidase enzyme of enzyme classification EC 1.10.3.1, a monophenol monooxygenase enzyme of enzyme classification EC 1.14.99.1, a bilirubin oxidase enzyme of enzyme classification EC 1.3.3.5, and an ascorbate oxidase enzyme of enzyme classification EC 1.10.3.3.  
     
     
         9 . The process of  claim 7  wherein said treating further includes a hydrolase.  
     
     
         10 . The process of  claim 9  wherein said hydrolase is a xylanase.  
     
     
         11 . The process of  claim 7  wherein said monomer, dimer, or oligomer is an organic soluble fraction of lignin or a lignin sulfonate.  
     
     
         12 . The process of  claim 7  wherein said derivative of cinnamic acid is selected from: 4-(dimethylamino)cinnamic acid; 3-hydroxy-4-methyoxycinnamic acid; 4-aminocinnamic acid; and 4-chlorocinnamic acid.  
     
     
         13 . The process of  claim 7  that further comprises adding an oxidizing agent.  
     
     
         14 . The process of  claim 13  wherein said oxidizing agent is at least one of air, oxygen, and hydrogen peroxide.  
     
     
         15 . A process for bleaching a lignin-containing material, comprising treating the material with an enzyme exhibiting laccase activity and an enzyme enhancing agent selected from: 
 (i) a monomer, dimer, or oligomer of at least one of:                          wherein:    R 1  is OCH 3 , H, or another moiety;    R 2  is OCH 3 , H, or another moiety;    R 3  is OH, sulfonate, or another moiety;    R 4  is an OH or methylene that connects to another monomer; and    R 5  is SO 3 H or H;    (ii) N-benzylidene benzylamine or a substituted N-benzylidene benzylamine having the structure:                          wherein:    R 6  is H, OCH 3 , NH(CH 3 ) 2 , Cl, Br, an aliphatic group, or an aromatic group; and    R 7  is H, OCH 3 , NH(CH 3 ) 2 , Cl, Br, an aliphatic group, or an aromatic group;    (iii) a derivative of cinnamic acid having the structure:                          wherein:    R 8  is OCH 3 , NH2, Cl, NH(CH 3 ) 2 ; and    R 9  is OH or H;    (vi) salicylic acid;    (v) epigallocatechin gallate;    (vi) melamine;    (vii) 3,4-dihydroxybenzyl alcohol or a substituted 3.4-dihydroxybenzyl alcohol;    (viii) a hardwood black liquor;    (ix) a hardwood black liquor that comprises (i) said monomer, dimer, or oligomer;    (x) a softwood black liquor; and    (xi) a softwood black liquor that comprises (i) said monomer, dimer, or oligomer.    
     
     
         16 . The process of  claim 15  wherein said enzyme exhibiting laccase activity is selected from a laccase enzyme of enzyme classification EC 1.10.3.2, a catechol oxidase enzyme of enzyme classification EC 1.10.3.1, a monophenol monooxygenase enzyme of enzyme classification EC 1.14.99.1, a bilirubin oxidase enzyme of enzyme classification EC 1.3.3.5, and an ascorbate oxidase enzyme of enzyme classification EC 1.10.3.3.  
     
     
         17 . The process of  claim 15  wherein said treating further includes a hydrolase.  
     
     
         18 . The process of  claim 17  wherein said hydrolase is a xylanase.  
     
     
         19 . The process of  claim 15  wherein said monomer, dimer, or oligomer is an organic soluble fraction of lignin or a lignin sulfonate.  
     
     
         20 . The process of  claim 15  wherein said derivative of cinnamic acid is selected from: 4-(dimethylamino)cinnamic acid; 3-hydroxy-4-methyoxycinnamic acid; 4-aminocinnamic acid; and 4-chlorocinnamic acid.  
     
     
         21 . The process of  claim 15  that further comprises adding an oxidizing agent.  
     
     
         22 . The process of  claim 21  wherein said oxidizing agent is at least one of air, oxygen, and hydrogen peroxide.  
     
     
         23 . The process of  claim 15  wherein said material is a wood pulp.  
     
     
         24 . The process of  claim 23  wherein said wood pulp is a raw material used to form a cellulose or a cellulose derivative.  
     
     
         25 . A process for enhancing the activity of an enzyme exhibiting laccase activity, comprising adding an enzyme enhancing agent to said enzyme, wherein said enzyme enhancing agent is selected from: 
 (i) a monomer, dimer, or oligomer of at least one of:                          wherein:    R 1  is OCH 3 , H, or another moiety;    R 2  is OCH 3 , H, or another moiety;    R 3  is OH, sulfonate, or another moiety;    R 4  is an OH or methylene that connects to another monomer; and    R 5  is SO 3 H or H;    (ii) N-benzylidene benzylamine or a substituted N-benzylidene benzylamine having the structure:                          wherein:    R 6  is H, OCH 3 , NH(CH 3 ) 2 , Cl, Br, an aliphatic group, or an aromatic group; and    R 7  is H, OCH 3 , NH(CH 3 ) 2 , Cl, Br, an aliphatic group, or an aromatic group;    (iii) a derivative of cinnamic acid having the structure:                          wherein:    R 8  is OCH 3 , NH2, Cl, NH(CH 3 ) 2 ; and    R 9  is OH or H;    (vi) salicylic acid;    (v) epigallocatechin gallate;    (vi) melamine;    (vii) 3,4-dihydroxybenzyl alcohol or a substituted 3.4-dihydroxylbenzyl alcohol;    (viii) a hardwood black liquor;    (ix) a hardwood black liquor that comprises (i) said monomer, dimer, or oligomer;    (x) a softwood black liquor; and    (xi) a softwood black liquor that comprises (i) said monomer, dimer, or oligomer.