Biofilms, components and methods of use to reduce biofouling and contamination

Abstract

Biofilms are provided which are capable of regulating their own thickness, reducing contamination and preventing biofouling. Constructs are introduced into bacteria that comprise nucleic acid molecules encoding an autoinducer synthase polypeptide, a transcriptional regulator and a biofilm dispersal protein. Nucleic acid molecules may also be introduced which encode a nitric oxide synthase, an epoxide hydrolase, or both. Biofilms of the bacteria may be used to reduce biofouling and contamination of a surface.

1 . A method of producing a living self-controlled biofilm of engineered bacteria cells on a surface, said method comprising:
 a) producing at least one engineered bacteria cell by introducing into said at least one bacterial cell a quorum sensing nucleic acid construct, comprising,
 i. a nucleic acid molecule encoding an autoinducer synthase polypeptide; 
 ii. a nucleic acid molecule encoding a transcriptional regulator, capable of being activated by said autoinducer synthase polypeptide; and 
 iii. a nucleic acid molecule encoding a biofilm dispersal protein to produce said at least one engineered bacteria cell comprising said construct; and 
   b) producing a self-controlled biofilm of said at least one engineered bacteria-cells, wherein said quorum sensing nucleic acid construct in response to activation of said transcriptional regulatory and production of said biofilm dispersal protein reduces the thickness of said self-controlled biofilm compared to biofilm not comprising said quorum sensing nucleic acid construct.   
     
     
         2 .- 4 . (canceled) 
     
     
         5 . The method of  claim 1 , further comprising introducing into said at least one bacterial cell a nucleic acid molecule encoding nitric oxide synthase. 
     
     
         6 . The method of  claim 1 , further comprising introducing into said bacterial cell a nucleic acid molecule encoding epoxide hydrolase. 
     
     
         7 .- 8 . (canceled) 
     
     
         9 . The method of  claim 1 , wherein said thickness of said biofilm is at least six fold less than said biofilm not comprising said quorum sensing nucleic acid construct. 
     
     
         10 . The method of  claim 1 , wherein said surface comprises a reverse osmosis membrane. 
     
     
         11 . The method of  claim 1 , wherein said at least one bacterial cell comprises a gram negative bacterial cell. 
     
     
         12 . A living self-controlled biofilm of engineered bacteria, said engineered bacteria comprising a nucleic acid construct comprising:
 a) a nucleic acid molecule encoding an autoinducer synthase polypeptide;   b) a nucleic acid molecule encoding a transcriptional regulator, capable of being activated by said auto inducer synthase polypeptide; and   c) a nucleic acid molecule encoding a biofilm dispersal protein to produce engineered bacteria comprising said construct and producing a self-controlled biofilm of said engineered bacteria.   
     
     
         13 .- 14 . (canceled) 
     
     
         15 . The biofilm of  claim 12 , further comprising a nucleic acid molecule encoding nitric oxide synthase. 
     
     
         16 . The biofilm of  claim 12 , further comprising a nucleic acid molecule encoding epoxide hydrolase. 
     
     
         17 . The biofilm of  claim 12 , further comprising a nucleic acid molecule encoding nitric oxide synthase and a nucleic acid molecule encoding epoxide hydrolase. 
     
     
         18 . A nucleic acid construct, comprising:
 a) a nucleic acid molecule encoding an autoinducer synthase polypeptide;   b) a nucleic acid molecule encoding a transcriptional regulator, capable of being activated by said auto inducer synthase polypeptide; and   c) a nucleic acid molecule encoding a biofilm dispersal protein.   
     
     
         19 . The nucleic acid construct of  claim 18 , wherein said autoinducer synthase comprises LasI and said transcriptional regulator comprises LasR, said biofilm dispersal protein comprises BdcA, and further comprising nucleic acid sequence encoding nitric oxide synthase and a nucleic acid molecule encoding epoxide hydrolase.