Microorganism having carbon dioxide fixation cycle introduced thereinto

Patent metadata

Field	Value
Publication number	US-9828618-B2
Application number	US-201414428928-A
Country	US
Kind code	B2
Filing date	Jan 23, 2014
Priority date	Jan 24, 2013
Publication date	Nov 28, 2017
Grant date	Nov 28, 2017

Abstract

An acetyl-CoA producing microorganism obtained by imparting at least one enzymatic activity selected from the group consisting of malate thiokinase, malyl-CoA lyase, glyoxylate carboligase, 2-hydroxy-3-oxopropionate reductase, and hydroxypyruvate reductase, to a microorganism that does not have any of the following (a), (b), (c), (d) or (e): (a) a carbon dioxide fixation cycle including an enzymatic reaction from malonyl-CoA to malonate semialdehyde or 3-hydroxypropionate; (b) a carbon dioxide fixation cycle including an enzymatic reaction from acetyl-CoA and CO 2 to pyruvate; (c) a carbon dioxide fixation cycle including an enzymatic reaction from crotonyl-CoA and CO 2 to ethylmalonyl-CoA or glutaconyl-CoA; (d) a carbon dioxide fixation cycle including an enzymatic reaction from CO 2 to formate; or (e) at least one selected from the group consisting of malate thiokinase and malyl-CoA lyase.

First claim

The invention claimed is:

1. A method of producing acetyl-CoA, comprising:
culturing an acetyl-CoA producing microorganism comprising an acetyl-CoA production cycle obtained by imparting at least one enzymatic activity selected from the group consisting of malate thiokinase, malyl-CoA lyase, glyoxylate carboligase, 2-hydroxy-3-oxopropionate reductase, and hydroxypyruvate reductase, to a microorganism that does not have any of the following (a), (b), (c), (d) or (e):
(a) a carbon dioxide fixation cycle including an enzymatic reaction from malonyl-CoA to malonate semialdehyde catalyzed by malonate semialdehyde dehydrogenase, or an enzymatic reaction from malonyl-CoA to 3-hydroxypropionate catalyzed by malonyl-CoA reductase;
(b) a carbon dioxide fixation cycle including an enzymatic reaction from acetyl-CoA and CO2 to pyruvate catalyzed by pyruvate synthase;
(c) a carbon dioxide fixation cycle including an enzymatic reaction from crotonyl-CoA and CO 2 to ethylmalonyl-CoA catalyzed by crotonyl-CoA carboxylase-reductase, or an enzymatic reaction from crotonyl-CoA and CO 2 to glutaconyl-CoA catalyzed by methylcrotonyl-CoA carboxylase;
(d) a carbon dioxide fixation cycle including an enzymatic reaction from CO2 to formate catalyzed by formate dehydrogenase; or
(e) at least one selected from the group consisting of malate thiokinase and malyl-CoA lyase,
the acetyl-CoA producing microorganism being obtained without imparting any of (a), (b), (c), or (d) to the microorganism, or such that the microorganism exhibits none of the functions of (a), (b), (c), or (d) even if one or more of (a), (b), (c), or (d) is imparted thereto; and
supplying at least one selected from the group consisting of a carbonate ion or a bicarbonate ion with a total supply amount of 150 mmol/L or more, and carbon dioxide gas with an average bubble diameter of 100 μm or more, to a culture medium used for the culturing
wherein the microorganism produces acetyl-CoA.

2. The method of producing acetyl-CoA according to claim 1 , further comprising resupplying carbon dioxide generated by the culturing to the culture medium used for the culturing.

3. The method of producing acetyl-CoA according to claim 1 ,
wherein the acetyl-CoA producing microorganism comprises an acetyl-CoA production cycle including the following (f), (g), (h), (i), (j), (k), (l), and (m):
(f) at least one selected from the group consisting of: pyruvate kinase and pyruvate carboxylase; phosphoenolpyruvate carboxylase; and phosphoenolpyruvate carboxykinase;
(g) malate dehydrogenase;
(h) malate thiokinase;
(i) malyl-CoA lyase;
(j) glyoxylate carboligase;
(k) at least one selected from the group consisting of: 2-hydroxy-3-oxopropionate reductase; and hydroxypyruvate isomerase and hydroxypyruvate reductase;
(l) at least one selected from the group consisting of: glycerate 2-kinase; and glycerate 3-kinase and phosphoglycerate mutase; and
(m) enolase.

4. The method of producing acetyl-CoA according to claim 1 , wherein the microorganism that does not have any of (a), (b), (c), (d), or (e), is a microorganism belonging to Enterobacteriaceae or a microorganism belonging to coryneform bacteria.

5. The method of producing acetyl-CoA according to claim 1 ,
wherein the microorganism that does not have any of (a), (b), (c), (d), or (e), is an Escherichia bacterium or a Pantoea bacterium belonging to Enterobacteriaceae, or is a Corynebacterium bacterium belonging to coryneform bacteria.

6. The method of claim 1 further comprising allowing the acetyl-CoA producing microorganism to produce isopropyl alcohol, using, as an intermediate, the acetyl-CoA produced.

7. The method of claim 1 further comprising allowing the acetyl-CoA producing microorganism to produce acetone, using, as an intermediate, the acetyl-CoA produced.

8. A method of producing glutamic acid, comprising:
culturing an acetyl-CoA producing microorganism comprising an acetyl-CoA production cycle obtained by imparting at least one enzymatic activity selected from the group consisting of malate thiokinase, malyl-CoA lyase, glyoxylate carboligase, 2-hydroxy-3-oxopropionate reductase, and hydroxypyruvate reductase, to a microorganism that does not have any of the following (a), (b), (c), (d) or (e) to produce acetyl CoA:
(a) a carbon dioxide fixation cycle including an enzymatic reaction from malonyl-CoA to malonate semialdehyde catalyzed by malonate semialdehyde dehydrogenase, or an enzymatic reaction from malonyl-CoA to 3-hydroxypropionate catalyzed by malonyl-CoA reductase;
(b) a carbon dioxide fixation cycle including an enzymatic reaction from acetyl-CoA and CO2 to pyruvate catalyzed by pyruvate synthase;
(c) a carbon dioxide fixation cycle including an enzymatic reaction from crotonyl-CoA and CO 2 to ethylmalonyl-CoA catalyzed by crotonyl-CoA carboxylase-reductase, or an enzymatic reaction from crotonyl-CoA and CO 2 to glutaconyl-CoA catalyzed by methylcrotonyl-CoA carboxylase;
(d) a carbon dioxide fixation cycle including an enzymatic reaction from CO2 to formate catalyzed by formate dehydrogenase; or
(e) at least one selected from the group consisting of malate thiokinase and malyl-CoA lyase,
the acetyl-CoA producing microorganism being obtained without imparting any of (a), (b), (c), or (d) to the microorganism, or such that the microorganism exhibits none of the functions of (a), (b), (c), or (d) even if one or more of (a), (b), (c), or (d) is imparted thereto;
supplying at least one selected from the group consisting of a carbonate ion or a bicarbonate ion with a total supply amount of 150 mmol/L or more, and carbon dioxide gas with an average bubble diameter of 100 μm or more, to a culture medium used for the culturing and
allowing the acetyl-CoA producing microorganism to produce glutamic acid, using the acetyl CoA.

9. A method of producing acetyl-CoA comprising:
culturing an acetyl-CoA producing microorganism comprising an acetyl-CoA production cycle obtained by imparting at least one enzymatic activity selected from the group consisting of malate thiokinase, malyl-CoA lyase, glyoxylate carboligase, 2-hydroxy-3-oxopropionate reductase, and hydroxypyruvate reductase, to a microorganism that does not have any of the following (a), (b), (c), (d) or (e):
(a) a carbon dioxide fixation cycle including an enzymatic reaction from malonyl-CoA to malonate semialdehyde catalyzed by malonate semialdehyde dehydrogenase, or an enzymatic reaction from malonyl-CoA to 3-hydroxypropionate catalyzed by malonyl-CoA reductase;
(b) a carbon dioxide fixation cycle including an enzymatic reaction from acetyl-CoA and CO2 to pyruvate catalyzed by pyruvate synthase;
(c) a carbon dioxide fixation cycle including an enzymatic reaction from crotonyl-CoA and CO 2 to ethylmalonyl-CoA catalyzed by crotonyl-CoA carboxylase-reductase, or an enzymatic reaction from crotonyl-CoA and CO 2 to glutaconyl-CoA catalyzed by methylcrotonyl-CoA carboxylase;
(d) a carbon dioxide fixation cycle including an enzymatic reaction from CO2 to formate catalyzed by formate dehydrogenase; or
(e) at least one selected from the group consisting of malate thiokinase and malyl-CoA lyase,
the acetyl-CoA producing microorganism being obtained without imparting any of (a), (b), (c), or (d) to the microorganism, or such that the microorganism exhibits none of the functions of (a), (b), (c), or (d) even if one or more of (a), (b), (c), or (d) is imparted thereto; and
supplying sodium sulfite with a total supply amount of from 0.01 g/l to 50 g/L to a culture medium used for the culturing wherein the microorganism produces acetyl-CoA.

10. The method of producing acetyl-CoA according to claim 1 , further comprising supplying sodium sulfite with

Assignees

• Mitsui Chemicals Inc

Inventors

• Hidesaki Tomonori
• Fujii Ryota
• Matsumoto Yoshiko
• Madhavan Anjali
• Chong Su Sun

Classifications

• C12N15/77

  for Corynebacterium; for Brevibacterium · CPC title

• C12P19/26

  Preparation of nitrogen-containing carbohydrates · CPC title

• C12P7/40

  containing a carboxyl group {including Peroxycarboxylic acids} · CPC title

• C12P13/14

  Glutamic acid; Glutamine · CPC title

• C12N9/0006

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