Compositions and methods useful in bioremediation of polychlorinated biphenyls

Abstract

The invention includes an isolated bacterium and a consortium of microorganisms including an isolated bacterium of the invention, which is capable of utilizing as a sole carbon source a polychlorinated biphenyl. The invention also includes a recombinant microorganism, such as a bacterium, a yeast or a bacteriophage, transfected with nucleic acid from an isolated bacterium of the invention, which is capable of utilizing as a sole carbon source a polychlorinated biphenyl. The invention also includes an isolated bacterium, which is capable of utilizing as a sole carbon source a chlorobenzoate. In addition, the invention includes an isolated nucleic acid and recombinant enzymes useful in the degradation of a polychlorinated biphenyl. The invention also includes methods for the bioremediation or enhancement of the bioremediation of a PCB-contaminated environment using the bacteria or microorganisms of the invention.

What is claimed is:  
     
         1 . An isolated bacterium which is capable of utilizing as a sole carbon source a polychlorinated biphenyl, wherein said polychlorinated biphenyl contains at least two chlorine atoms associated therewith.  
     
     
         2 . The isolated bacterium of  claim 1 , wherein said polychlorinated biphenyl is 2,3-dichloro-biphenyl.  
     
     
         3 . The isolated bacterium of  claim 1 , wherein said polychlorinated biphenyl is 3,4-dichloro-biphenyl.  
     
     
         4 . The isolated bacterium of  claim 1 , wherein said polychlorinated biphenyl is 2,2′-dichloro-biphenyl.  
     
     
         5 . The isolated bacterium of  claim 1 , wherein said polychlorinated biphenyl is 2,4′-dichloro-biphenyl.  
     
     
         6 . The isolated bacterium of  claim 2 , wherein said bacterium is strain SK-1.  
     
     
         7 . The isolated bacterium of  claim 3 , wherein said bacterium is strain SK-2.  
     
     
         8 . The isolated bacterium of  claim 4 , wherein said bacterium is strain SK-4.  
     
     
         9 . The isolated bacterium of  claim 5 , wherein said bacterium is strain SK-3.  
     
     
         10 . The isolated bacterium of  claim 5 , wherein said bacterium is strain SK-4.  
     
     
         11 . An isolated bacterium which is capable of utilizing as a sole carbon source a monochloro-biphenyl.  
     
     
         12 . The isolated bacterium of  claim 1   1 , wherein said monochloro-biphenyl is selected from the group consisting of 2-chlorobiphenyl, 3-chlorobiphenyl and 4-chlorobiphenyl.  
     
     
         13 . An isolated bacterium which is capable of utilizing as a sole carbon source 4-chlorobenzoate.  
     
     
         14 . An isolated bacterium which is capable of utilizing both 4-chlorobenzoate and a mono-chlorobiphenyl, individually, as a sole carbon source.  
     
     
         15 . The isolated bacterium of  claim 13 , wherein said bacterium is strain SK-3.  
     
     
         16 . A consortium of microorganisms comprising a bacterium selected from the group consisting of SK-1, SK-2, SK-3 and SK-4.  
     
     
         17 . A recombinant bacterium, which is capable of utilizing as a sole carbon source a polychlorinated biphenyl, wherein said polychlorinated biphenyl contains at least two chlorine atoms associated therewith.  
     
     
         18 . A recombinant yeast, which is capable of utilizing as a sole carbon source a polychlorinated biphenyl, wherein said polychlorinated biphenyl contains at least two chlorine atoms associated therewith.  
     
     
         19 . A recombinant bacteriophage, comprising nucleic acid which when expressed in a bacterium infected with said bacteriophage confers on said bacterium the ability to utilize as a sole carbon source a polychlorinated biphenyl, wherein said polychlorinated biphenyl contains at least two chlorine atoms associated therewith.  
     
     
         20 . An isolated nucleic acid encoding an enzyme for degrading a polychlorinated biphenyl, wherein said nucleic acid is obtained from an isolated bacterium which is capable of utilizing as a sole carbon source a polychlorinated biphenyl, wherein said polychlorinated biphenyl contains at least two chlorine atoms associated therewith.  
     
     
         21 . A recombinant enzyme for degrading a polychlorinated biphenyl wherein said recombinant enzyme is encoded by nucleic acid obtained from the genome of an isolated bacterium which is capable of utilizing as a sole carbon source a polychlorinated biphenyl, wherein said polychlorinated biphenyl contains at least two chlorine atoms associated therewith.  
     
     
         22 . The recombinant enzyme of  claim 21 , wherein said enzyme is expressed in a bacterium.  
     
     
         23 . The recombinant enzyme of  claim 21 , wherein said enzyme is expressed in a yeast.  
     
     
         24 . The recombinant enzyme of  claim 21 , wherein said recombinant enzyme is substantially pure.  
     
     
         25 . A method for the bioremediation of a PCB-contaminated environment, said method comprising 
 a) adding an isolated bacterium to said PCB-contaminated environment, wherein said bacterium is capable of utilizing as a sole carbon source a polychlorinated biphenyl, wherein said polychlorinated biphenyl contains at least two chlorine atoms associated therewith, and    b) incubating said bacterium in said environment for a period of time sufficient to permit degradation of a polychlorinated biphenyl in said environment, thereby bioremediating said environment.    
     
     
         26 . A method for enhancing the bioremediation of a PCB-contaminated environment, said method comprising 
 a) adding an isolated bacterium to said PCB-contaminated environment, wherein said bacterium is capable of utilizing as a sole carbon source a polychlorinated biphenyl, wherein said polychlorinated biphenyl contains at least two chlorine atoms associated therewith, and    b) incubating said bacterium in said environment for a period of time sufficient to permit degradation of a polychlorinated biphenyl in said environment, thereby enhancing the bioremediation of said environment.    
     
     
         27 . A method for the bioremediation of a PCB-contaminated environment, said method comprising 
 a) adding an isolated bacterium to said PCB-contaminated environment, wherein said bacterium is capable of utilizing as a sole carbon source a polychlorinated biphenyl, wherein said polychlorinated biphenyl contains at least two chlorine atoms associated therewith;    b) adding a cosubstrate selected from the group consisting of biphenyl and napthalene to said PCB-contaminated environment whereby cometabolism of said cosubstrate and a PCB may occur, and    c) incubating said bacterium in said environment for a period of time sufficient to permit degradation of a polychlorinated biphenyl in said environment, thereby bioremediating said environment.    
     
     
         28 . A method for the bioremediation of a PCB-contaminated environment, said method comprising 
 a) adding a recombinant enzyme for degrading a polychlorinated biphenyl to said PCB-contaminated environment, wherein said recombinant enzyme is obtained from an isolated bacterium which is capable of utilizing as a sole carbon source a polychlorinated biphenyl, wherein said polychlorinated biphenyl contains at least two chlorine atoms associated therewith, and    b) incubating said recombinant enzyme in said PCB-contaminated environment for a period of time sufficient to permit degradation of a polychlorinated biphenyl, thereby bioremediating said environment.    
     
     
         29 . A method for the bioremediation of PCB-contaminated environment, said method comprising 
 a) adding to said PCB-contaminated environment, a recombinant microorganism selected from the group consisting of a bacterium, a yeast, and a bacteriophage, wherein said recombinant microorganism comprises a nucleic acid encoding an enzyme for degrading a polychlorinated biphenyl, wherein said nucleic acid is obtained from an isolated bacterium which is capable of utilizing as a sole carbon source a polychlorinated biphenyl, wherein said polychlorinated biphenyl contains at least two chlorine atoms associated therewith, and    b) incubating said recombinant microorganism in said PCB-contaminated environment for a period of time sufficient to permit degradation of a polychlorinated biphenyl, thereby bioremediating said environment.    
     
     
         30 . A method for the bioremediation of a PCB-contaminated environment, said method comprising 
 a) adding an isolated bacterium to said PCB-contaminated environment, wherein said bacterium is capable of utilizing as a sole carbon source a polychlorinated biphenyl, wherein said polychlorinated biphenyl contains at least two chlorine atoms associated therewith;    b) adding an isolated bacterium to said PCB-contaminated environment, wherein said bacterium is capable of utilizing a chlorobenzoate as a sole carbon source, and    c) incubating said bacteria in said environment for a period of time sufficient to permit degradation of a polychlorinated biphenyl and a chlorobenzoate in said environment, thereby bioremediating said environment.