IPC분류정보
국가/구분 |
United States(US) Patent
등록
|
국제특허분류(IPC7판) |
|
출원번호 |
US-0891760
(2010-09-27)
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등록번호 |
US-8809027
(2014-08-19)
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발명자
/ 주소 |
- Lynch, Michael D
- Mercogliano, Christopher P.
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출원인 / 주소 |
- OPX Biotechnologies, Inc.
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대리인 / 주소 |
Wilson Sonsini Goodrich & Rosati
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인용정보 |
피인용 횟수 :
8 인용 특허 :
66 |
초록
▼
Microorganism compositions are described that comprise combinations of genetic modifications that include a genetic modification to increase oxaloacetate alpha-decarboxylase enzymatic activity. By such genetic modification a 3-hydroxypropionic acid (“3-HP”) production pathway is provided or improved
Microorganism compositions are described that comprise combinations of genetic modifications that include a genetic modification to increase oxaloacetate alpha-decarboxylase enzymatic activity. By such genetic modification a 3-hydroxypropionic acid (“3-HP”) production pathway is provided or improved. In various embodiments, comprising other genetic modifications, including selected gene disruptions, 3-HP production is greater than in a control microorganism lacking such combinations of genetic modifications.
대표청구항
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1. A genetically modified microorganism comprising: i) a heterologous nucleic acid sequence encoding a 3 hydroxypropionate dehydrogenase;ii) a heterologous nucleic acid sequence encoding a cyanase or a carbonic anhydrase; andiii) a heterologous nucleic acid sequence encoding an NADPH-dependent trans
1. A genetically modified microorganism comprising: i) a heterologous nucleic acid sequence encoding a 3 hydroxypropionate dehydrogenase;ii) a heterologous nucleic acid sequence encoding a cyanase or a carbonic anhydrase; andiii) a heterologous nucleic acid sequence encoding an NADPH-dependent transhydrogenase or an acetyl-CoA carboxylase. 2. The genetically modified microorganism of claim 1, wherein the 3-hydroxypropionate dehydrogenase is a native or mutated form of a mmsB protein or a native or mutated form of a ydfG protein. 3. The genetically modified microorganism of claim 1, wherein said genetically modified microorganism comprises a heterologous nucleic acid sequence encoding an NADPH-dependent transhydrogenase. 4. The genetically modified microorganism of claim 1, further comprising a modification of a gene to increase the NADPH/NADP+ ratio, wherein the modification is selected from the group consisting of: increasing activity of pgi, increasing activity of pntAB, gapA:gapN substitution or replacement, and disrupting sthA. 5. The genetically modified microorganism of claim 1, wherein said genetically modified microorganism comprises a heterologous nucleic acid sequence encoding a cyanase or a carbonic anhydrase that increases intracellular bicarbonate levels. 6. The genetically modified microorganism of claim 1, wherein said genetically modified microorganism comprises a heterologous nucleic acid sequence encoding an acetyl-CoA carboxylase pathway. 7. The genetically modified microorganism of claim 1, wherein said genetically modified microorganism is selected from the group consisting of: Escherichia coli, Cupriavidus necator, Oligotropha carboxidovorans, and Pseudomonas putida. 8. The genetically modified microorganism of claim 1, wherein said genetically modified microorganism is an Escherichia coli. 9. The genetically modified microorganism of claim 1, wherein said genetically modified microorganism is a gram-positive bacterium. 10. The genetically modified microorganism of claim 1, wherein said genetically modified microorganism is selected from the group consisting of: Clostridium, Rhodococcus, Bacillus, Lactobacillus, Enterococcus, Paenibacillus, Arthrobacter, Corynebacterium, and Brevibacterium. 11. The genetically modified microorganism of claim 1, wherein said genetically modified microorganism is selected from the group consisting of: Bacillus licheniformis, Paenibacillus macerans, Rhodococcus erythropolis, Lactobacillus plantarum, Enterococcus faecium, Enterococcus gallinarium, Enterococcus faecalis, and Bacillus subtilis. 12. The genetically modified microorganism of claim 1, wherein said genetically modified microorganism is a fungus. 13. The genetically modified microorganism of claim 1, wherein said genetically modified microorganism is a yeast. 14. The genetically modified microorganism of claim 13, wherein said yeast is selected from the group consisting of: Pichia, Candida, Hansenula, and Saccharomyces. 15. The genetically modified microorganism of claim 1, wherein said genetically modified microorganism is selected from the group consisting of: Zymomonas, Escherichia, Pseudomonas, Alcaligenes, Salmonella, Shigella, Burkholderia, Oligotropha, and Klebsiella. 16. The genetically modified microorganism of claim 1, further comprising at least one heterologous nucleic acid sequence encoding an oxaloacetate alpha-decarboxylase. 17. The genetically modified microorganism of claim 16, additionally comprising at least one heterologous nucleic acid sequence encoding a phosphoenolpyruvate carboxykinase or a phosphoenol pyruvate carboxylase. 18. The genetically modified microorganism of claim 17, further comprising at least one genetic modification to reduce enzymatic activity of a protein selected from the group consisting of: lactate dehydrogenase, pyruvate formate lyase, phosphate acetyltransferase, heat stable, histidyl phosphorylatable protein, phosphoryl transfer protein, polypeptide chain, pyruvate kinase I, and pyruvate kinase II. 19. The genetically modified microorganism of claim 16, wherein the oxaloacetate alpha-decarboxylase is selected from the group consisting of: SEQ ID NOs: 54, 55, and 56. 20. A genetically modified microorganism comprising: i) a heterologous nucleic acid sequence encoding an enzyme selected from the group consisting of:a native or mutated form of a mmsB protein, and a native or mutated form of a ydfG protein,ii) a genetic modification to decrease or eliminate an enzymatic activity selected from the group consisting of: lactate dehydrogenase, phosphate acetyltransferase, and pyruvate-formate lyase;iii) a heterologous nucleic acid sequence encoding an acetyl-CoA carboxylase; andiv) a heterologous nucleic acid sequence encoding a cyanase. 21. The genetically modified microorganism of claim 20, wherein the lactate dehydrogenase is encoded by a nucleic acid sequence having at least 90% sequence identity to SEQ ID NO:19 or has an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 20. 22. The genetically modified microorganism of claim 20, wherein the phosphate acetyltransferase is encoded by a nucleic acid sequence having at least 90% sequence identity to SEQ ID NOs: 25 or has an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 26. 23. The genetically modified microorganism of claim 20, wherein the pyruvate-formate lyase is encoded by a nucleic acid sequence having at least 90% sequence identity to SEQ ID NOs: 21 or has an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 22. 24. The genetically modified microorganism of claim 20, wherein said genetically modified microorganism comprises a heterologous nucleic acid sequence encoding an acetyl-CoA carboxylase. 25. A genetically modified microorganism comprising: i) a heterologous nucleic acid sequence encoding a 3-hydroxypropionate dehydrogenase; andii) a heterologous nucleic acid sequence encoding a cyanase or a carbonic anhydrase. 26. The genetically modified microorganism of claim 25, further comprising: iii) a heterologous nucleic acid sequence encoding an acetyl-CoA carboxylase.
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