Modified bacterium useful for producing an organic molecule
The present invention provides for a genetically modified host cell or bacterium capable of producing an organic molecule, wherein the bacterium is capable of using hydrogen sulfide as an electron donor, carbon dioxide (CO 2 ) as a carbon source, and/or nitrate as an electron acceptor. The present invention is useful in the wastewater treatment industry, in particular in municipal wastewater treatment plants (WWTP).
1. A genetically modified bacterium capable of producing an organic molecule comprising a heterologous nucleic acid encoding a ‘tesA integrated into a chromosome of the genetically modified bacterium, wherein the genetically modified bacterium is capable of using hydrogen sulfide as an electron donor, carbon dioxide (CO 2 ) as a source, and nitrate as an electron acceptor, and the organic molecule is heterologous or the genetically modified bacterium overproduces the organic molecule compared to an unmodified bacterium; wherein the organic molecule is a fatty acid and the genetically modified bacterium is a chemolithoautotroph, wherein the organic molecule is a methyl ketone and the heterologous nucleic acid encodes two or more of acyl-CoA oxidase, FadB, FadM, and FadD.
2. The bacterium of claim 1 , wherein the acyl-CoA oxidase a Micrococcus luteus acyl-CoA oxidase, and FadB, FadM, and FadD are Escherichia coli FadB, FadM, and FadD.
3. The genetically modified bacterium of claim 1 , wherein the heterologous nucleic acid encodes one or more of pyruvate kinase and pyruvate dehydrogenase, and acetyl-CoA carboxylase.
4. The genetically modified bacterium of claim 1 , wherein the genetically modified bacterium is a species from the genera Thiobacillus, Thiomicrospira, Sulfurimonas, Thioalkalivibrio , or Sulfurovum.
5. The genetically modified bacterium of claim 4 , wherein the genetically modified bacterium is a Thiobacillus denitrificans.
6. The bacterium of claim 4 , wherein the bacterium is a Sulfurimonas denitrificans.
7. The genetically modified bacterium of claim 1 comprising one or more genes encoding enzymes for using hydrogen sulfide as an electron donor, CO 2 as a carbon source, and/or nitrate as an electron acceptor.
8. The genetically modified bacterium of claim 7 , wherein the genetically modified bacterium uses hydrogen sulfide as a sole electron donor and/or CO 2 as a sole carbon source.
9. The genetically modified bacterium of claim 7 , wherein the organic molecule is a biofuel.
10. A method of growing the genetically modified bacterium of claim 1 , comprising:
(a) providing the genetically modified bacterium of claim 1 , and
(b) culturing the genetically modified bacterium in a liquid comprising hydrogen sulfide and CO 2 , and optionally nitrate,
whereby optionally the genetically modified bacterium produces the organic molecule that the genetically unmodified bacterium does not produce or overproduces the organic molecule when compared to an unmodified bacterium.
11. The method of claim 10 , wherein the liquid is a wastewater.
12. The method of claim 10 , wherein the providing step comprises constructing the genetically modified bacterium.
13. The method of claim 12 , wherein the constructing step comprises introducing heterologous nucleic acid encoding one or more enzymes for using sulfide as an electron donor, carbon dioxide (CO 2 ) as a carbon source, and/or nitrate as an electron acceptor into an unmodified bacterium.
14. The method of claim 13 , wherein the introduced heterologous nucleic acid is capable of stable maintenance in the genetically modified bacterium.
15. The method of claim 10 , wherein the culturing step results in reducing the amount of hydrogen sulfide, CO 2 , and/or nitrate in the liquid.
16. The method of claim 10 , wherein the culturing step is a continuous culture wherein additional liquid optionally comprising further hydrogen sulfide, CO 2 , and/or nitrate is added to the liquid comprising the genetically modified bacterium.