Compositions and methods for the biosynthesis of 1,4-butanediol and its precursors
US-2015368676-A1 · Dec 24, 2015 · US
US9587256B2 · US · B2
| Field | Value |
|---|---|
| Publication number | US-9587256-B2 |
| Application number | US-201314426290-A |
| Country | US |
| Kind code | B2 |
| Filing date | Sep 6, 2013 |
| Priority date | Sep 6, 2012 |
| Publication date | Mar 7, 2017 |
| Grant date | Mar 7, 2017 |
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Provided herein are genetically engineered microbes that include at least a portion of a carbon fixation pathway, and in one embodiment, use molecular hydrogen to drive carbon dioxide fixation. In one embodiment, the genetically engineered microbe is modified to convert acetyl CoA, molecular hydrogen, and carbon dioxide to 3-hydroxypropionate, 4-hydroxybutyrate, acetyl CoA, or the combination thereof at levels greater than a control microbe. Other products may also be produced. Also provided herein are cell free compositions that convert acetyl CoA, molecular hydrogen, and carbon dioxide to 3-hydroxypropionate, 4-hydroxybutyrate, acetyl CoA, or the combination thereof. Also provided herein are methods of using the genetically engineered microbes and the cell free compositions.
Opening claim text (preview).
What is claimed is: 1. A genetically engineered microbe modified to convert acetyl CoA, molecular hydrogen, and carbon dioxide to 3-hydroxypropionate, wherein the 3-hydroxypropionate is produced at an increased level compared to a control microbe, wherein the microbe is a hyperthermophile, wherein the microbe is a member of the domain Archaea or a member of the domain Bacteria, and wherein the microbe comprises an exogenous coding region encoding a polypeptide, wherein the polypeptide has an activity selected from acetyl/propionyl-CoA carboxylase activity, malonyl/succinyl-CoA reductase activity, and malonate semialdehyde reductase activity. 2. The genetically engineered microbe of claim 1 wherein the hyperthermophile is a member of the Order Thermococcales, a member of the Order Sulfolobales, or a member of the Order Thermotogales. 3. The genetically engineered microbe of claim 1 wherein the microbe further comprises a NADPH-dependent hydrogenase. 4. The genetically engineered microbe of claim 1 wherein the microbe comprises exogenous coding regions encoding subunits of the NADPH-dependent hydrogenase. 5. The genetically engineered microbe of claim 1 wherein the exogenous coding region is operably linked to a temperature sensitive promoter, to a constitutive promoter, or to a non-regulated promoter. 6. The genetically engineered microbe of claim 2 wherein the hyperthermophile is Thermococcus kodakarensis, T. onnurineus, Sulfolobus solfataricus, S. islandicus, S. Acidocaldarius , or Pyrococcus furiosus. 7. The genetically engineered microbe of claim 1 wherein the hyperthermophile is a Caldicellulosiruptor spp. 8. The genetically engineered microbe of claim 7 wherein the Caldicellulosiruptor is C. bescii. 9. A method comprising incubating the genetically engineered microbe of claim 1 under anaerobic conditions suitable for converting acetyl CoA, molecular hydrogen, and carbon dioxide to 3-hydroxypropionate. 10. The method of claim 9 further comprising recovering the 3-hydroxypropionate.
3-Hydroxypropionyl-CoA synthase (6.2.1.36) · CPC title
Methylmalonyl-CoA mutase (5.4.99.2) · CPC title
Methylmalonyl-CoA epimerase (5.1.99.1) · CPC title
3-Hydroxypropionyl-CoA dehydratase (4.2.1.116) · CPC title
Acrylyl-CoA reductase (NADH) (1.3.1.95) · CPC title
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