Extraction and purification of phosphodiesterase

Abstract

A process for purifying a phosphodiesterase 1 (PDE-1) from a cell including heating an extract of a cell formed from a solution including at least on divalent cation, to increase the specific activity of PDE-1 in the extract.

1 . A process for purifying a phosphodiesterase 1 (PDE-1) from a cell including heating an extract of a cell formed from a solution including at least one divalent cation, to increase the specific activity of PDE-1 in the extract, wherein the concentration of the divalent cation is at least about 20 mM.  
   
   
       2 . A process according to  claim 1  wherein the concentration of the divalent cation is about 20 mM to 50 mM.  
   
   
       3 . A process according to  claim 2  wherein the concentration of the divalent cation is about 25 mM.  
   
   
       4 . A process according to  claim 2  wherein the concentration of the divalent cation is about 30 mM.  
   
   
       5 . A process according to  claim 2  wherein the concentration of the divalent cation is about 35 mM.  
   
   
       6 . A process according to  claim 2  wherein the concentration of the divalent cation is about 40 mM.  
   
   
       7 . A process according to  claim 2  wherein the concentration of the divalent cation is about 45 mM.  
   
   
       8 . A process according to  claim 1  wherein the divalent cation is magnesium or calcium.  
   
   
       9 . A process for purifying PDE-1 from a barley cell including: 
 releasing PDE-1 from the cell into a solution including at least about 20 mM calcium and at least about 20 mM magnesium to form an extract; and    heating the extract to increase the specific activity of PDE-1 in the extract.    
   
   
       10 . A process according to  claim 9  wherein the extract is maintained in conditions for promoting solubilisation of the phosphodiesterase in the extract prior to heating the extract.  
   
   
       11 . A process according to  claim 10  wherein the extract is maintained at less than 10° C.  
   
   
       12 . A process according to  claim 11  comprising the further step of: 
 utilising chromatography to purify PDE-1 from the heated extract.    
   
   
       13 . A process according to  claim 12  wherein anion exchange chromatography is utilised to purify PDE-1 from the heated extract.