Method for enhancing enzyme activity at elevated temperature

Abstract

A method for enhancing activity of enzyme at an elevated temperature which comprises adding a substance exhibiting chaperone function such as a saccharide to a reaction mixture containing the enzyme. The method can improve activity of enzymes more easily and more effectively and hence afford increased enzyme activity at an elevated temperature.

What is claimed is:  
     
         1 . A method for enhancing activity of enzyme at an elevated temperature which comprises adding a substance exhibiting chaperone function to a reaction mixture containing the enzyme.  
     
     
         2 . The method of  claim 1 , wherein the reaction mixture is at a temperature of 45 to 110° C.  
     
     
         3 . The method of  claim 1 , wherein the substance exhibiting chaperone function is one or more substances selected from the group consisting of saccharides, polyalcohols, amino acids and their derivatives, and chaperone proteins.  
     
     
         4 . The method of  claim 3 , wherein the saccharide is one or more saccharides selected from the group consisting of trehalose, maltose, glucose, sucrose, lactose, xylobiose, agarobiose, cellobiose, levanbiose, quitobiose, 2-β-glucuronosylglucuronic acid, allose, altrose, galactose, gulose, idose, mannose, talose, sorbitol, levulose, xylitol and arabitol.  
     
     
         5 . The method of  claim 4 , wherein the saccharide is trehalose. sorbitol, levulose, xylitol or arabitol.  
     
     
         6 . The method of  claim 3 , wherein the amino acid or derivative thereof is one or more members selected from the group consisting of N e -acetyl-β-lysine, alanine, γ-aminobutyric acid, betain, N α -carbamoyl-L-glutamine 1-amide, choline, dimethylthetine, ecotine, glutamate, β-glutammine, glycine, octopine, proline, sarcosine, taurine and trymethylamine N-oxide.  
     
     
         7 . The method of  claim 6 , wherein the amino acid or derivative thereof is betain or sarconsine.  
     
     
         8 . The method of  claim 3 , wherein the chaperone protein is selected from those of Thermophiric bacteria and heat shock proteins.  
     
     
         9 . The method of  claim 1 , wherein one or more polyalcohols are added to the reaction mixture.  
     
     
         10 . The method of  claim 1 , wherein the enzyme is a polymerase or a restriction enzyme.  
     
     
         11 . The method of  claim 10 , wherein the polymerase is a reverse transcriptase or DNA polymerase.