Method for activating denatured protein

Abstract

A method for activating a denatured protein, includes the steps of: adding a detergent to the denatured protein to allow a protein-detergent complex to be formed; and adding high-molecular weight amylose to the protein-detergent complex so that the high-molecular weight amylose removes the detergent.

What is claimed is:  
     
         1 . A method for activating a denatured protein, comprising the steps of: 
 adding a detergent to the denatured protein to allow a protein-detergent complex to be formed; and    adding high-molecular weight amylose to the protein-detergent complex so that the high-molecular weight amylose removes the detergent.    
     
     
         2 . A method according to  claim 1 , 
 further comprising the step of adding a denaturing agent to the denatured protein to cause the denatured protein to be in an unfolded state,    wherein the step of adding the denaturing agent is performed before the step of adding the detergent to the denatured protein.    
     
     
         3 . A method according to  claim 1 , wherein the high-molecular weight amylose is an inclusion compound which includes a lower alcohol.  
     
     
         4 . A method according to  claim 3 , wherein the lower alcohol is butanol.  
     
     
         5 . A method according to  claim 1 , wherein the high-molecular weight amylose has an average molecular weight in the range of about 10,000 to about 20,000,000.  
     
     
         6 . A method according to  claim 5 , wherein the high-molecular weight amylose has an average molecular weight in the range of about 20,000 to about 11,000,000.  
     
     
         7 . A method according to  claim 1 , wherein the high-molecular weight amylose is an amylose which is synthesized by using an enzyme.  
     
     
         8 . A method according to  claim 1 , wherein the detergent is selected from the group consisting of: polyoxyethylene sorbitan ester, polyoxyethylene dodecyl ether, polyoxyethylene fatty acid ester, sucrose fatty acid ester, cetyltrimethylammonium bromide, sodium deoxycholate, hexadecyltrimethylammonium bromide, and myristylsulfobetaine.  
     
     
         9 . A method according to  claim 1 , wherein the denatured protein includes an α helix structural region.  
     
     
         10 . A method according to  claim 1 , wherein the denatured protein includes a β sheet structural region.  
     
     
         11 . A method according to  claim 1 , wherein the denatured protein includes an intramolecular S—S bond.  
     
     
         12 . A method for activating a denatured protein including an, α helix structural region, comprising the steps of: 
 adding an excess of a polyoxyethylene type detergent to the denatured protein to allow a protein-detergent complex to be formed, thereby preventing aggregation of the protein; and  
 adding high-molecular weight amylose to the protein-detergent complex so that the high-molecular weight amylose removes the detergent, thereby allowing the protein to be folded again into a higher-order structure for acquiring activity, thus activating the protein.  
 
     
     
         13 . A method according to  claim 12 , wherein the polyoxyethylene type detergent is selected from the group consisting of: polyoxyethylene sorbitan ester, polyoxyethylene dodecyl ether, polyoxyethylene fatty acid ester, and sucrose fatty acid ester.  
     
     
         14 . A method for activating a denatured protein including a β sheet structural region, comprising the steps of: 
 adding an excess of an ionic type detergent to the denatured protein to allow a protein-detergent complex to be formed, thereby preventing aggregation of the protein; and  
 adding high-molecular weight amylose to the protein-detergent complex so that the high-molecular weight amylose removes the detergent, thereby allowing the protein to be folded again into a higher-order structure for acquiring activity, thus activating the protein.  
 
     
     
         15 . A method for activating a denatured protein including an intramolecular S—S bond, comprising the steps of: 
 adding an excess of an ionic type detergent to the denatured protein to allow a protein-detergent complex to be formed, thereby preventing aggregation of the protein; and  
 adding high-molecular weight amylose to the protein-detergent complex so that the high-molecular weight amylose removes the detergent, thereby allowing the protein to be folded again into a higher-order structure for acquiring activity, thus activating the protein.  
 
     
     
         16 . A method according to  claim 14 , wherein the ionic type detergent is selected from the group consisting of: cetyltrimethylammonium bromide, sodium deoxycholate, hexadecyltrimethylammonium bromide, and myristylsulfobetaine.  
     
     
         17 . A method according to  claim 15 , wherein the ionic type detergent is selected from the group consisting of: cetyltrimethylammonium bromide, sodium deoxycholate, hexadecyltrimethylammonium bromide, and myristylsulfobetaine.  
     
     
         18 . A kit for refolding a protein, comprising at least one kind of high-molecular weight amylose and at least one kind of detergent.  
     
     
         19 . A kit according to  claim 18 , wherein the high-molecular weight amylose is an inclusion compound which includes a lower alcohol.  
     
     
         20 . A kit according to  claim 19 , wherein the lower alcohol is butanol.  
     
     
         21 . A kit according to  claim 18 , wherein the high-molecular weight amylose has an average molecular weight in the range of about 10,000 to about 20,000,000.  
     
     
         22 . A kit according to  claim 21 , wherein the high-molecular weight amylose has an average molecular weight in the range of about 20,000 to about 11,000,000.  
     
     
         23 . A kit according to  claim 18 , wherein the high-molecular weight amylose is an amylose which is synthesized by using an enzyme.  
     
     
         24 . A kit according to  claim 18 , wherein the detergent is selected from the group consisting of: polyoxyethylene sorbitan ester, polyoxyethylene dodecyl ether, polyoxyethylene fatty acid ester, sucrose fatty acid ester, cetyltrimethylammonium bromide, sodium deoxycholate, hexadecyltrimethylammonium bromide, and myristylsulfobetaine.