Sequestration of carbon dioxide with hydrogen to useful products

Patent metadata

Field	Value
Publication number	US-2017226542-A1
Application number	US-201715410205-A
Country	US
Kind code	A1
Filing date	Jan 19, 2017
Priority date	Sep 6, 2012
Publication date	Aug 10, 2017
Grant date	—

Abstract

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.

First claim

1 - 2 . (canceled)

3 . A genetically engineered microbe modified to convert acetyl CoA, molecular hydrogen and carbon dioxide to 4-hydroxybutyrate, wherein the 4-hydroxybutyrate is produced at increased levels compared to a control microbe, wherein the microbe is a hyperthermophile, and wherein the microbe comprises an exogenous coding region encoding a polypeptide, wherein the polypeptide has an activity selected from 3-hydroxypropionate:CoA ligase activity, 3-hydroxypropionyl-CoA dehydratase activity, acryloyl-CoA reductase activity, methylmalonyl-CoA epimerase activity, methylmalonyl-CoA mutase activity, and succinate semialdehyde reductase activity.

4 - 6 . (canceled)

7 . The genetically engineered microbe of claim 3 wherein the hyperthermophile is an archeon.

8 . The genetically engineered microbe of claim 7 wherein the archeon is a member of the Order Thermococcales, a member of the Order Sulfolobales, or a member of the Order Thermotogales.

9 . The genetically engineered microbe of claim 8 wherein the archeon is Thermococcus kodakarensis, T. onnurineus, Sulfolobus solfataricus, S. islandicus, S. acidocaldarius , or Pyrococcus furiosus.

10 . (canceled)

11 . The genetically engineered microbe of claim 3 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.

12 - 14 . (canceled)

15 . The genetically engineered microbe of claim 3 wherein the microbe produces 4-hydroxybutyrate, and wherein the microbe comprises an exogenous coding region encoding a polypeptide, wherein the polypeptide has an activity selected from 4-hydroxybutyrate:CoA ligase activity, 4-hydroxybutyrl-CoA dehydratase activity, crotonyl-CoA hydratase/(S)-3-hydroxybutyrl-CoA dehydrogenase activity, and acetoacetyl-CoA β-ketothiolase activity.

16 . (canceled)

17 . The genetically engineered microbe of claim 3 wherein an exogenous coding region is operably linked to a temperature sensitive promoter, to a constitutive promoter, or to a non-regulated promoter.

18 . The genetically engineered microbe of claim 3 wherein the microbe further comprises a hydrogenase.

19 . The genetically engineered microbe of claim 18 wherein the hydrogenase is a NADPH-dependent hydrogenase.

20 . The genetically engineered microbe of claim 19 wherein the microbe comprises exogenous coding regions encoding subunits of the NADPH-dependent hydrogenase.

21 . The genetically engineered microbe of claim 20 wherein the subunits of the NADPH-dependent hydrogenase comprise a hydrogenase alpha subunit and a hydrogenase delta subunit.

22 . The genetically engineered microbe of claim 21 wherein the subunits of the NADPH-dependent hydrogenase further comprise a hydrogenase beta subunit and a hydrogenase gamma subunit.

23 - 24 . (canceled)

25 . A method comprising incubating the genetically engineered microbe of claim 3 under anaerobic conditions suitable for converting acetyl CoA, molecular hydrogen, and carbon dioxide to 4-hydroxybutyrate.

26 . The method of claim 25 further comprising recovering the 4-hydroxybutyrate.

27 - 29 . (canceled)

30 . The method of claim 25 wherein the incubating comprises an incubation temperature of at least 75° C.

31 - 33 . (canceled)

34 . A cell free composition that converts acetyl CoA, molecular hydrogen and carbon dioxide to 4-hydroxybutyrate, wherein the composition comprises a polypeptide having 3-hydroxypropionate:CoA ligase activity, a polypeptide having 3-hydroxypropionyl-CoA dehydratase activity, a polypeptide having acryloyl-CoA reductase activity, a polypeptide having methylmalonyl-CoA epimerase activity, a polypeptide having methylmalonyl-CoA mutase activity, and a polypeptide having succinate semialdehyde reductase activity.

35 . A cell free method for fixing CO 2 comprising incubating the cell free composition of claim 34 under anaerobic conditions suitable for the fixation of CO 2 by the conversion of acetyl CoA, molecular hydrogen and carbon dioxide to 4-hydroxybutyrate.

36 . The cell free method of claim 35 further comprising isolating the 4-hydroxybutyrate.

37 - 40 . (canceled)

41 . The cell free composition of claim 34 wherein the cell free composition further comprises a polypeptide comprising acetyl/propionyl-CoA carboxylase activity, a polypeptide comprising malonyl/succinyl-CoA reductase activity, a polypeptide comprising malonate semialdehyde reductase activity, and a polypeptide comprising NADPH-dependent hydrogenase activity.

Assignees

• Univ Georgia
• Univ North Carolina State

Inventors

• Adams Michael W W
• Kelly Robert M
• Hawkins Aaron B
• Menon Angeli Lal
• Lipscomb Gina Lynette Pries
• Schut Gerrit Jan

Classifications

• C12N9/0006

  acting on CH-OH groups as donors (1.1) · CPC title

• C12Y602/01036

  3-Hydroxypropionyl-CoA synthase (6.2.1.36) · CPC title

• C12Y103/01095

  Acrylyl-CoA reductase (NADH) (1.3.1.95) · CPC title

• C12Y504/99002

  Methylmalonyl-CoA mutase (5.4.99.2) · CPC title

• C12Y101/01

  with NAD+ or NADP+ as acceptor (1.1.1) · CPC title