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

Patent metadata

Field	Value
Publication number	US-10172362-B2
Application number	US-201615334168-A
Country	US
Kind code	B2
Filing date	Oct 25, 2016
Priority date	Oct 26, 2015
Publication date	Jan 8, 2019
Grant date	Jan 8, 2019

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.

First claim

What is claimed is:

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 bacteria cell a quorum sensing nucleic acid construct, comprising,
i. a nucleic acid molecule encoding an autoinducer synthase polypeptide comprising Lasl;
ii. a nucleic acid molecule encoding a transcriptional regulator comprising LasR capable of being activated by said autoinducer synthase polypeptide; and
iii. a nucleic acid molecule encoding a biofilm dispersal protein comprising BdcA 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 regulator 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. The method of claim 1 , further comprising introducing into said at least one bacteria cell a nucleic acid molecule encoding nitric oxide synthase.

3. The method of claim 1 , further comprising introducing into said bacteria a nucleic acid molecule encoding epoxide hydrolase.

4. 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.

5. The method of claim 1 , wherein said surface comprises a reverse osmosis membrane.

6. The method of claim 1 , wherein said at least one bacteria cell comprises a gram negative bacteria cell.

7. A living self-controlled biofilm of engineered bacteria, said engineered bacteria comprising a nucleic acid construct comprising:
a) a nucleic acid molecule encoding an auto inducer synthase polypeptide comprising LasI;
b) a nucleic acid molecule encoding a transcriptional regulator comprising LasR, capable of being activated by said auto inducer synthase polypeptide; and
c) a nucleic acid molecule encoding a biofilm dispersal protein comprising BdcA to produce engineered bacteria comprising said construct and producing a self-controlled biofilm of said engineered bacteria.

8. The biofilm of claim 7 , further comprising a nucleic acid molecule encoding nitric oxide synthase.

9. The biofilm of claim 7 , further comprising a nucleic acid molecule encoding epoxide hydrolase.

10. The biofilm of claim 7 , further comprising a nucleic acid molecule encoding nitric oxide synthase and a nucleic acid molecule encoding epoxide hydrolase.

Assignees

• Penn State Res Found

Inventors

• Wood Thomas K
• Kumar Manish
• Wood Thammajun L

Classifications

• C12N1/04

  Preserving or maintaining viable microorganisms (immobilised microorganisms C12N11/00) · CPC title

• C12N1/20

  Bacteria; Culture media therefor · CPC title

• C12N15/74

  Vectors or expression systems specially adapted for prokaryotic hosts other than E. coli, e.g. Lactobacillus, Micromonospora · CPC title

• C12N9/14

  Hydrolases (3) · CPC title

• C12Y303/02003

  Epoxide hydrolase (3.3.2.3) (C12Y303/02009, C12Y303/0201 take precedence) · CPC title