Conversion of compactin to pravastatin by actinomadura

Abstract

A method for converting compactin to pravastatin is described. Compactin is provided and contacted with an agent derived from Actinomadura under conditions in which the agent converts compactin to pravastatin. Also described are an Actinomadura strain, an Actinomadura cell free extract, an Actinomadura hydroxylase, and a method for lowering cholesterol levels in mammals.

What is claimed is:  
     
         1 . A method for converting compactin to pravastatin, comprising: 
 providing said compactin; and    contacting said compactin with an agent derived from Actinomadura under conditions in which said agent converts compactin to pravastatin.    
     
     
         2 . The method of  claim 1  further comprising the step of isolating said pravastatin.  
     
     
         3 . The method of  claim 1  wherein said agent is a hydroxylation enzyme produced by said Actinomadura.  
     
     
         4 . The method of  claim 3  wherein said hydroxylation enzyme is a hydroxylase that is cytochrome P450 system-independent.  
     
     
         5 . The method of  claim 3  wherein said hydroxylation enzyme is constitutive.  
     
     
         6 . The method of  claim 1  wherein said compactin is provided by providing a microorganism that produces said compactin.  
     
     
         7 . The method of  claim 6  wherein said microorganism is selected from the group consisting of a fungus and a bacterium.  
     
     
         8 . The method of  claim 1  wherein said compactin is provided by providing a cell free extract of a microorganism that produces said compactin.  
     
     
         9 . The method of  claim 1  wherein said compactin is provided by providing cell free culture media from a pregrown culture of a microorganism that produces said compactin.  
     
     
         10 . The method of  claim 1  wherein said compactin is provided by providing a solution comprising compactin.  
     
     
         11 . The method of  claim 1  wherein said compactin is provided by providing substantially purified compactin.  
     
     
         12 . The method of  claim 1 , wherein said compactin is provided in shots.  
     
     
         13 . The method of  claim 1  wherein said compactin is contacted with said agent by contacting whole cells of said Actinomadura with said compactin.  
     
     
         14 . The method of  claim 1  wherein said compactin is contacted with said agent by contacting a cell free extract of said Actinomadura with said compactin.  
     
     
         15 . The method of  claim 1  wherein said compactin is contacted with said agent by contacting cell free culture media from a pregrown culture of said Actinomadura with said compactin.  
     
     
         16 . The method of  claim 1  wherein said compactin is contacted with said agent by contacting a solution comprising said agent of said Actinomadura with said compactin.  
     
     
         17 . The method of  claim 6  wherein said compactin is contacted with said agent by contacting a pregrown culture of said microorganism that produces said compactin with a pregrown culture of said Actinomadura.  
     
     
         17 . The method of  claim 6  wherein said compactin is contacted with said agent by contacting a pregrown culture of said microorganism that produces said compactin with a starting culture of said Actinomadura.  
     
     
         19 . The method of  claim 6  wherein said compactin is contacted with said agent by contacting a starting culture of said microorganism that produces said compactin with a pregrown culture of said Actinomadura.  
     
     
         20 . The method of  claim 6  wherein said compactin is contacted with said agent by contacting a starting culture of said microorganism that produces said compactin with a starting culture of said Actinomadura.  
     
     
         21 . The method of  claim 6  wherein said compactin is contacted with said agent by contacting a cell free extract of said microorganism that produces said compactin with a culture of said Actinomadura.  
     
     
         22 . The method of  claim 6  wherein said compactin is contacted with said agent by contacting a culture of said microorganism that produces said compactin with a cell free extract of said Actinomadura.  
     
     
         23 . The method of  claim 6  wherein said compactin is contacted with said agent by contacting a cell free extract of said microorganism that produces said compactin with a cell free extract of said Actinomadura.  
     
     
         24 . The method of  claim 1 , wherein conversion of said compactin to said pravastatin is at least about 50%.  
     
     
         25 . The method of  claim 1 , wherein conversion of said compactin to said pravastatin is at least about 80%.  
     
     
         26 . A cell free extract derived from Actinomadura, comprising an agent that converts compactin to pravastatin.  
     
     
         27 . The cell free extract of  claim 26  wherein said cell free extract is derived from ruptured cells of said Actinomadura from which particulate cellular debris has been essentially removed.  
     
     
         28 . The cell free extract of  claim 26  wherein said agent is a hydroxylase.  
     
     
         29 . The cell free extract of  claim 28  wherein said hydroxylase is a constitutive enzyme, wherein the enzymatic activity of said hydroxylase is stimulated by any of ATP, ascorbic acid and Mg ++ , but not stimulated by any of Fe ++  and Fe +++ , and wherein said hydroxylase is cytochrome P450 system-independent.  
     
     
         30 . The cell free extract of  claim 26  wherein conversion of said compactin to said pravastatin is at least about 50%.  
     
     
         31 . The cell free extract of  claim 26  wherein conversion of said compactin to said pravastatin is at least about 80%.  
     
     
         32 . A hydroxylase from Actinomadura that converts compactin to pravastatin, 
 said hydroxylase being a constitutive enzyme;    the activity of said hydroxylase being stimulated by any of ATP, ascorbic acid and Mg ++ , but not by Fe ++  or Fe +++ ; and    said hydroxylase being cytochrome P450 system-independent.    
     
     
         33 . Purified Actinomadura 55678, comprising an agent for converting compactin to pravastatin.  
     
     
         34 . The Actinomadura of  claim 33  wherein said agent is a hydroxylase.  
     
     
         35 . A method for treating a mammal to lower the blood cholesterol level of the mammal, comprising: 
 providing pravastatin,    said pravastatin being derived from compactin by contacting said compactin with an agent derived from Actinomadura that converts compactin to pravastatin; and    administering said pravastatin to a mammal in need of such treatment to cause a lower blood cholesterol level in the mammal.    
     
     
         36 . The method of  claim 35  wherein said agent is a hydroxylase.