Microorganism with modified aldehyde:ferredoxin oxidoreductase activity and related methods

Patent metadata

Field	Value
Publication number	US-2017327849-A1
Application number	US-201715594252-A
Country	US
Kind code	A1
Filing date	May 12, 2017
Priority date	May 14, 2016
Publication date	Nov 16, 2017
Grant date	—

Abstract

The invention provides a non-naturally occurring bacterium having decreased or eliminated activity of an enzyme that catalyzes the reaction defined by EC 1.2.7.5, such as aldehyde:ferredoxin oxidoreductase (AOR). Optionally, the bacterium also has decreased or eliminated activity of an enzyme that catalyzes the reaction defined by EC 1.2.1.10 and/or EC 1.1.1.1, such as aldehyde dehydrogenase, alcohol dehydrogenase, or bifunctional aldehyde/alcohol dehydrogenase. The invention further provides methods of producing products by culturing the bacterium in the presence of a gaseous substrate containing one or more of CO, CO 2 , and H 2 .

First claim

1 . A non-naturally occurring bacterium having decreased or eliminated activity of an enzyme that catalyzes the reaction defined by EC 1.2.7.5 compared to a parental bacterium.

2 . The non-naturally occurring bacterium of claim 1 , wherein the non-naturally occurring bacterium comprises at least one disruptive mutation in a gene encoding the enzyme that catalyzes the reaction defined by EC 1.2.7.5.

3 . The non-naturally occurring bacterium of claim 1 , wherein the enzyme that catalyzes the reaction defined by EC 1.2.7.5 is aldehyde:ferredoxin oxidoreductase.

4 . The non-naturally occurring bacterium of claim 1 , wherein the non-naturally occurring bacterium further has decreased or eliminated activity of at least one enzyme that catalyzes the reaction defined by EC 1.2.1.10 and/or EC 1.1.1.1 compared to the parental bacterium.

5 . The non-naturally occurring bacterium of claim 4 , wherein the non-naturally occurring bacterium comprises at least one disruptive mutation in a gene encoding the enzyme that catalyzes the reaction defined by EC 1.2.1.10 and/or EC 1.1.1.1.

6 . The non-naturally occurring bacterium of claim 4 , wherein the enzyme that catalyzes the reaction defined by EC 1.2.1.10 and/or EC 1.1.1.1 is selected from the group consisting of bifunctional aldehyde/alcohol dehydrogenase, aldehyde dehydrogenase, and alcohol dehydrogenase.

7 . The non-naturally occurring bacterium of claim 1 , wherein the non-naturally occurring bacterium produces a product selected from the group consisting of acetyl-CoA, acetoacetyl-CoA, acetoacetate, acetone, isopropanol, 3-hydroxyisovaleryl-CoA, 3-hydroxyisovalerate, isobutylene, isoprene, 3-hydroxybutyryl-CoA, 3-hydroxybutyrate, 3-hydroxybutyrylaldehyde, 1,3-butanediol, 2-hydroxyisobutyryl-CoA, 2-hydroxyisobutyrate, pyruvate, acetolactate, acetoin, 2,3-butanediol and lactate.

8 . The non-naturally occurring bacterium of claim 1 , wherein the non-naturally occurring bacterium consumes a gaseous substrate comprising one or more of CO, CO 2 , and H 2 .

9 . The non-naturally occurring bacterium of claim 1 , wherein the parental bacterium is selected from the group consisting of Alkalibaculum bacchi, Blautia product, Butyribacterium methylotrophicum, Chloroflexus aurantiacus, Clostridium aceticum, Clostridium acetobutylicum, Clostridium autoethanogenum, Clostridium botulinum, Clostridium carboxidivorans, Clostridium coskatii, Clostridium drakei, Clostridium formicoaceticum, Clostridium ljungdahlii, Clostridium ragsdalei, Desulfovibrio vulgaris, Eubacterium limosum, Geobacter sulfurreducens, Methylomicrobium alcaliphilum, Moorella thermoautrophica, Moorella thermoacetica, Rhodospirillum rubrum, Sporomusa ovata, Sporomusa silvacetica, Sporomusa sphaeroides, Thermanaerovibrio acidaminovorans, Thermanaerovibrio acidaminovorans, Thermoanaerobacter wiegelii, Thermodesulfovibrio yellowstonii, Thermodesulfovibrio yellowstonii , and Thermus thermophilus.

10 . A method of producing a product by culturing the non-naturally occurring bacterium of claim 1 in the presence of a gaseous substrate comprising one or more of CO, CO 2 , and H 2 .

11 . The method of claim 10 , wherein the non-naturally occurring bacterium comprises at least one disruptive mutation in a gene encoding the enzyme that catalyzes the reaction defined by EC 1.2.7.5.

12 . The method of claim 10 , wherein the enzyme that catalyzes the reaction defined by EC 1.2.7.5 is aldehyde:ferredoxin oxidoreductase.

13 . The method of claim 10 , wherein the non-naturally occurring bacterium further has decreased or eliminated activity of at least one enzyme that catalyzes the reaction defined by EC 1.2.1.10 and/or EC 1.1.1.1 compared to the parental bacterium.

14 . The method of claim 13 , wherein the non-naturally occurring bacterium comprises at least one disruptive mutation in a gene encoding the enzyme that catalyzes the reaction defined by EC 1.2.1.10 and/or EC 1.1.1.1.

15 . The method of claim 13 , wherein the enzyme that catalyzes the reaction defined by EC 1.2.1.10 and/or EC 1.1.1.1 is selected from the group consisting of bifunctional aldehyde/alcohol dehydrogenase, aldehyde dehydrogenase, and alcohol dehydrogenase.

16 . The method of claim 10 , wherein the product is an acetyl-CoA-derived product selected from the group consisting of acetyl-CoA, acetoacetyl-CoA, acetoacetate, acetone, isopropanol, 3-hydroxyisovaleryl-CoA, 3-hydroxyisovalerate, isobutylene, isoprene, 3-hydroxybutyryl-CoA, 3-hydroxybutyrate, 3-hydroxybutyrylaldehyde, 1,3-butanediol, 2-hydroxyisobutyryl-CoA, 2-hydroxyisobutyrate, pyruvate, acetolactate, acetoin, 2,3-butanediol and lactate.

17 . The method of claim 10 , wherein the parental bacterium is selected from the group consisting of Alkalibaculum bacchi, Blautia product, Butyribacterium methylotrophicum, Chloroflexus aurantiacus, Clostridium aceticum, Clostridium acetobutylicum, Clostridium autoethanogenum, Clostridium botulinum, Clostridium carboxidivorans, Clostridium coskatii, Clostridium drakei, Clostridium formicoaceticum, Clostridium ljungdahlii, Clostridium ragsdalei, Desulfovibrio vulgaris, Eubacterium limosum, Geobacter sulfurreducens, Methylomicrobium alcaliphilum, Moorella thermoautrophica, Moorella thermoacetica, Rhodospirillum rubrum, Sporomusa ovata, Sporomusa silvacetica, Sporomusa sphaeroides, Thermanaerovibrio acidaminovorans, Thermanaerovibrio acidaminovorans, Thermoanaerobacter wiegelii, Thermodesulfovibrio yellowstonii, Thermodesulfovibrio yellowstonii , and Thermus thermophilus.

Assignees

• Lanzatech Inc

Inventors

• Liew Fungmin
• Koepke Michael

Classifications

• C12Y102/0101

  Acetaldehyde dehydrogenase (acetylating) (1.2.1.10) · CPC title

• C12N1/20 Primary

  Bacteria; Culture media therefor · CPC title

• C12P7/065 Primary

  with microorganisms other than yeasts · CPC title

• C12Y102/07005

  Aldehyde ferredoxin oxidoreductase (1.2.7.5) · CPC title

• C12P5/007

  containing one or more isoprene units, i.e. terpenes (carotenes C12P23/00) · CPC title