Genetically engineered microorganisms and methods of use
This disclosure relates to genetically engineered microorganisms for treating or reducing the risk of bacterial infections or dysbiosis, and further discloses methods of making and using such microorganisms.
1. A genetically engineered microorganism, wherein the microorganism comprises:
(a) a microcin operon, and
(b) a controllable promoter for the microcin operon, wherein the microcin operon comprises one or more microcin genes, and the controllable promoter controls a level of expression of the one or more microcin genes, thereby controlling the amount of microcin produced by the genetically engineered microorganism, and wherein either or both of the microcin operon and the controllable promoter are heterologous to the microorganism, and wherein the controllable promoter is a pBAD or Pttr promoter.
2. The genetically engineered microorganism of claim 1 , wherein the genetically engineered microorganism is a bacterium.
3. The genetically engineered microorganism of claim 1 , wherein the genetically engineered microorganism is Escherichia coli.
4. The genetically engineered microorganism of claim 3 , wherein the E. coli is E. coli Nissle 1917 (EcN) or E. coli NGF-19.
5. The genetically engineered microorganism of claim 1 , wherein the microcin operon comprises one or more Microcin H47 (MccH47) genes.
6. The genetically engineered microorganism of claim 5 , wherein the microcin operon comprises mchB, mchC, and mchD.
7. The genetically engineered microorganism of claim 5 , wherein the microcin operon comprises mchB, mchC, mchD, mchE, mchF, mchX and mchI.
8. The genetically engineered microorganism of claim 5 , wherein the microorganism comprises mchA.
9. The genetically engineered microorganism of claim 1 , wherein the microcin operon comprises Microcin M genes or Microcin J25 genes.
10. The genetically engineered microorganism of claim 9 , wherein the microcin operon comprises mchS1 and mchS4 genes.
11. The genetically engineered microorganism of claim 1 , wherein the controllable promoter is a pBAD promoter.
12. The genetically engineered microorganism of claim 1 , wherein the microorganism further comprises a ttrBCA operon.
13. The genetically engineered microorganism of claim 1 , wherein the microcin operon and the controllable promoter are in the genome of the microorganism.
14. The genetically engineered microorganism of claim 1 , wherein the microcin operon and the controllable promoter are in a vector.
15. A vector comprising:
(a) a set of microcin genes, and
(b) a controllable promoter, wherein the controllable promoter is capable of controlling the expression level of at least one microcin gene, wherein the controllable promoter is a pBAD or Pttr promoter.
16. The vector of claim 15 , wherein the set of microcin genes comprises one or more Microcin H47 (MccH47) genes.
17. The vector of claim 16 , wherein the set of microcin genes comprises mchA, mchB, mchC, and mchD.
18. The vector of claim 16 , wherein the set of microcin genes comprises mchA, mchB, mchC, mchD, mchE, mchF, mchX and mchI.
19. The vector of claim 15 , wherein the set of microcin genes comprises Microcin M genes or Microcin J25 genes.
20. The vector of claim 19 , wherein the set of microcin genes comprises mchS1 and mchS4.
21. The vector of claim 15 , wherein the controllable promoter is a pBAD promoter.
22. The vector of claim 15 , wherein the vector further comprises an operon comprising one or more of a ttrBCA, a ttrC, and a ttrA.
23. The vector of claim 15 , wherein the vector is a plasmid.
24. A method of treating a subject for intestinal dysbiosis or a bacterial infection, the method comprising:
identifying a subject as having intestinal dysbiosis or a bacterial infection; and
administering to the subject a therapeutically effective amount of a composition comprising the genetically engineered microorganism of claim 1 .
25. The genetically engineered microorganism of claim 1 , wherein the controllable promoter is a Pttr promoter.
26. The genetically engineered microorganism of claim 15 , wherein the controllable promoter is a Pttr promoter.
27. The method of claim 24 , wherein the composition is administered orally.
28. The method of claim 24 , wherein the method comprises treating a bacterial infection.
29. The method of claim 28 , wherein the bacterial infection is a gram-negative bacterial infection.
30. The method of claim 24 , wherein the method comprises treating intestinal dysbiosis.