Compositions and methods for 3-hydroxypropionate bio-production from biomass
Methods of obtaining mutant nucleic acid sequences that demonstrate elevated oxaloacetate α-decarboxylase activity are provided. Compositions, such as genetically modified microorganisms that comprise such mutant nucleic acid sequences, are described, as are methods to obtain the same.
1. A recombinant bacterium transformed with at least one exogenous nucleic acid that encodes 3-hydroxypropanoic acid (3-HP) pathway enzymes, wherein said 3-HP pathway enzymes comprise a 2-keto acid decarboxylase and a dehydrogenase, and wherein production of 3-HP is enhanced as compared to a bacterium without the exogenous nucleic acid.
2. The recombinant bacterium of claim 1 , further comprising an exogenous nucleic acid that encodes a phosphoenolpyruvate carboxykinase.
3. The recombinant bacterium of claim 1 , wherein at least one exogenous nucleic acid is a heterologous nucleic acid.
4. The recombinant bacterium of claim 1 , wherein said bacterium is in an anaerobic culture medium.
5. The recombinant bacterium of claim 1 , wherein said dehydrogenase converts malonate semialdehyde to 3-HP.
6. The recombinant bacterium of claim 1 , wherein said dehydrogenase is a 3-hydroxypropionate dehydrogenase that converts malonate semialdehyde to 3-HP.
7. A method for producing 3-HP, comprising culturing the recombinant bacterium of claim 1 under conditions and for a sufficient period of time to produce 3-HP.
8. The method of claim 7 , wherein at least one exogenous nucleic acid is a heterologous nucleic acid.
9. The method of claim 7 , wherein said bacterium is in an anaerobic culture medium.
10. The method of claim 7 , wherein said dehydrogenase converts malonate semialdehyde to 3-HP.
11. The method of claim 7 , wherein said dehydrogenase is a 3-hydroxypropionate dehydrogenase that converts malonate semialdehyde to 3-HP.
12. A recombinant bacterium comprising a pathway modified for enhanced production of 3-hydroxypropionic acid (3-HP), wherein said bacterium is transformed with at least one heterologous nucleic acid that encodes enzymes of a 3-HP pathway, said enzymes comprise an oxaloacetate alpha-oxo-decarboxylase and a dehydrogenase, thereby enhancing a 3-HP bio-production process in said bacterium.
13. The recombinant bacterium of claim 12 , further comprising a heterologous nucleic acid that expresses the enzymatic activity of phosphoenolpyruvate carboxykinase for the pathway.
14. The recombinant bacterium of claim 12 , wherein at least one heterologous nucleic acid is a foreign nucleic acid.
15. The recombinant bacterium of claim 12 , wherein the bio-production of 3-HP is performed under anaerobic conditions.
16. The recombinant bacterium of claim 12 , wherein the bio-production of 3-HP is performed under microaerobic conditions.
17. The recombinant bacterium of claim 12 , wherein the dehydrogenase converts malonate semialdehyde to 3-HP.
18. The recombinant bacterium of claim 12 , wherein the dehydrogenase is 3-hydroxypropionate dehydrogenase.
19. A method for enhanced production of 3-hydroxypropionic acid (3-HP), the method comprising culturing the recombinant bacterium of claim 12 under suitable bio-production conditions for a suitable time to obtain enhanced bio-production of 3-HP.
20. The method of claim 19 , wherein at least one heterologous nucleic acid is a foreign nucleic acid.
21. The method of claim 19 , wherein the bio-production of 3-HP is performed under aerobic conditions.
22. The method of claim 19 , wherein the bio-production of 3-HP is performed under anaerobic conditions.
23. The method of claim 19 , wherein the bio-production of 3-HP goes through a malonate semialdehyde intermediate.
24. The method of claim 19 , wherein the dehydrogenase is 3-hydroxypropionate dehydrogenase.
25. A recombinant bacterium comprising a 3-hydroxypropionic acid (3-HP) pathway, wherein said bacterium is transformed with a heterologous nucleic acid that encodes 3-HP pathway enzymes and enhances a 3-HP bio-production process in said bacterium, wherein said 3-HP pathway enzymes comprise an oxaloacetate alpha-oxo-decarboxylase and a dehydrogenase, and wherein the bio-production of 3-HP is performed under microaerobic conditions.
26. A recombinant Escherichia coli bacterium, comprising a 3-hydroxypropionic acid (3-HP) pathway, wherein said bacterium is transformed with a heterologous nucleic acid that encodes 3-HP pathway enzymes, said 3-HP pathway enzymes comprise a 2-keto acid decarboxylase and a dehydrogenase and wherein production of 3-HP is enhanced as compared to a bacterium without the heterologous nucleic acid.