Methods, compositions and devices utilizing structurally stable cyanuric acid hydrolase

Abstract

The present invention relates to stable cyanuric acid hydrolase enzymes, compositions, and devices for use in the treatment of a liquid, such as water. The present invention also relates to methods of using these enzymes, compositions and devices for the treatment of a liquid, such as water.

What is claimed is: 
     
         1 . An isolated or purified structurally stable cyanuric acid hydrolase (CAH) enzyme. 
     
     
         2 . The CAH of  claim 1  having a K m  value for cyanuric acid of 25-150 μM. 
     
     
         3 . The CAH of  claim 1  having a k cat  value for cyanuric acid of 4.8-76 s −1 . 
     
     
         4 . The CAH of  claim 1 , wherein the enzyme is thermostable. 
     
     
         5 . The CAH of  claim 1 , wherein the enzyme retains at least 30% enzymatic activity at a temperature above 25° C. 
     
     
         6 . The CAH of  claim 1 , wherein the enzyme comprises amino acid sequence TEGNG(C/G/L)(V/M/A)ND(Y/F)(T/S)R (SEQ ID NO:15). 
     
     
         7 . The CAH of  claim 1 , wherein the enzyme comprises amino acid sequence (M/F/L/I)(V/I/M)(M/F/W)SGG (D/E/G/P)G(V/I/L/G/A)(L/I/M/A)(S/T/A/C)PHX(T/I/L/S)(V/I/L)(F/I/V) (SEQ ID NO:17), wherein X is any amino acid. 
     
     
         8 . The CAH of  claim 1 , wherein the enzyme comprises amino acid sequence TEGNGCVNDFTR (SEQ ID NO:16) and amino acid sequence FIMSGGEGVMTPHTVF (SEQ ID NO:18). 
     
     
         9 . The CAH of  claim 1 , wherein the enzyme is 350-380 amino acids in length. 
     
     
         10 . The CAH of  claim 1 , wherein the enzyme has a pI of about 5-6. 
     
     
         11 . The CAH of  claim 1 , wherein the enzyme is from  Moorella thermoacetica.    
     
     
         12 . The CAH of  claim 11 , wherein the enzyme is from  Moorella thermoacetica  ATCC 39073. 
     
     
         13 . The CAH of  claim 1 , wherein the enzyme has a specific activity of 12-18 μmmol/min/mg with cyanuric acid as substrate and less than 2 μmol/min/mg with barbituric acid as substrate. 
     
     
         14 . A composition for remediation of a liquid comprising the CAH of  claim 1  and polyethylene glycol (PEG) and/or KCl. 
     
     
         15 . The composition of  claim 14 , wherein the PEG is PEG4000. 
     
     
         16 . The composition of  claim 14 , wherein the PEG is present at a concentration of 5-50% by weight. 
     
     
         17 . The composition of  claim 14 , wherein KCl is present at a concentration of 50-500 mM. 
     
     
         18 . The composition of  claim 14 , wherein the liquid is water. 
     
     
         19 . A device for remediation of a liquid comprising a matrix and the stable cyanuric acid hydrolase of  claim 1 . 
     
     
         20 . The device of  claim 19 , wherein the device further comprises a casing or housing for the matrix, wherein water flows through the at least one casing and contacts the enzyme. 
     
     
         21 . The device of  claim 19 , further comprising a permeable layer.