Bacteria engineered to treat a disease or disorder
Genetically programmed microorganisms, such as bacteria, pharmaceutical compositions thereof, and methods of modulating and treating a disease and/or disorder are disclosed.
1. A genetically-engineered non-pathogenic bacterium for use in metabolizing a substrate of interest, the bacterium comprising;
at least one heterologous gene encoding a transporter for importing said substrate, and
at least one heterologous gene encoding a polypeptide involved in metabolizing said substrate,
wherein the at least one heterologous gene encoding the transporter for importing said substrate, and the at least one heterologous gene encoding the polypeptide involved in metabolizing said substrate are each operably linked to an inducible promoter that is induced by low oxygen or anaerobic conditions.
2. The genetically engineered non-pathogenic bacterium of claim 1 , wherein the inducible promoter operably linked to the at least one heterologous gene encoding the transporter and the inducible promoter operably linked to the at least one heterologous gene encoding the polypeptide involved in metabolizing the substrate are separate copies of the same promoter.
3. The genetically engineered non-pathogenic bacterium of claim 1 , wherein the at least one heterologous gene encoding the transporter and the at least one heterologous gene encoding the polypeptide involved in metabolizing the substrate are both operably linked to a single promoter.
4. The genetically engineered non-pathogenic bacterium of claim 1 , wherein the promoter operably linked to the at least one heterologous gene encoding the transporter for importing the substrate and the promoter operably linked to the at least one gene encoding the polypeptide involved in metabolizing the substrate are selected from the group consisting of an FNR-responsive promoter, an ANR-responsive promoter, and a DNR-responsive promoter.
5. The genetically engineered non-pathogenic bacterium of claim 1 , wherein the at least one heterologous gene encoding the transporter for importing the substrate is located on a chromosome in the bacterium.
6. The genetically engineered non-pathogenic bacterium of claim 1 , wherein the at least one heterologous gene encoding the transporter for importing the substrate is located on a plasmid in the bacterium.
7. The genetically engineered non-pathogenic bacterium of claim 1 , wherein the at least one heterologous gene encoding the polypeptide involved in metabolizing the substrate is located on a plasmid in the bacterium.
8. The genetically engineered non-pathogenic bacterium of claim 1 , wherein the at least one heterologous gene encoding the polypeptide involved in metabolizing the substrate is located on a chromosome in the bacterium.
9. The genetically engineered non-pathogenic bacterium of claim 1 , wherein the bacterium is a probiotic bacterium.
10. The genetically engineered non-pathogenic bacterium of claim 9 , wherein the bacterium is selected from the group consisting of Bacteroides, Bifidobacterium, Clostridium, Escherichia, Lactobacillus , and Lactococcus.
11. The genetically engineered non-pathogenic bacterium of claim 10 , wherein the bacterium is Escherichia coli strain Nissle.
12. The genetically engineered non-pathogenic bacterium of claim 1 , wherein the bacterium is an auxotroph in diaminopimelic acid or an enzyme in the thymidine biosynthetic pathway.
13. The genetically engineered non-pathogenic bacterium of claim 1 , wherein the bacterium is further engineered to harbor at least one heterologous gene encoding a substance toxic to the bacterium, wherein the at least one heterologous gene encoding the substance that is toxic to the bacterium is under the control of a promoter that is directly or indirectly induced by arabinose.
14. A pharmaceutically acceptable composition comprising the bacterium of claim 1 ; and a pharmaceutically acceptable carrier.
15. The pharmaceutically acceptable composition of claim 14 formulated for oral administration.
16. The genetically engineered non-pathogenic bacterium of claim 11 , wherein the bacterium is an auxotroph in diaminopimelic acid or an enzyme in the thymidine biosynthetic pathway.
17. The genetically engineered non-pathogenic bacterium of claim 11 , wherein the bacterium is further engineered to harbor at least one heterologous gene encoding a substance toxic to the bacterium, wherein the at least one heterologous gene encoding the substance that is toxic to the bacterium is under the control of a promoter that is directly or indirectly induced by arabinose.
18. A pharmaceutically acceptable composition comprising the bacterium of claim 11 ; and a pharmaceutically acceptable carrier.
19. A pharmaceutically acceptable composition comprising the bacterium of claim 16 ; and a pharmaceutically acceptable carrier.
20. A pharmaceutically acceptable composition comprising the bacterium of claim 17 ; and a pharmaceutically acceptable carrier.