Microorganism having novel acrylic acid synthesis pathway and method of producing acrylic acid by using the microorganism

Patent metadata

Field	Value
Publication number	US-9506089-B2
Application number	US-201514620049-A
Country	US
Kind code	B2
Filing date	Feb 11, 2015
Priority date	May 14, 2014
Publication date	Nov 29, 2016
Grant date	Nov 29, 2016

Abstract

A microorganism capable of producing an acrylic acid (AA), wherein activities of a pathway producing AA through conversions of 3-HP to 3-HP-CoA and 3-HP-CoA to AA-CoA in the microorganisms are increased; as well as a method of producing the microorganism and a method of producing an acrylic acid by using the same.

First claim

What is claimed is:

1. A genetically engineered microorganism that produces acrylate, wherein the genetically engineered microorganism comprises:
an exogenous polynucleotide encoding a coenzyme A (CoA) transferase catalyzing conversion of 3-hydroxypropionic acid (3-HP) to 3-hydroxypropionyl-CoA (3-HP-CoA) and belonging to EC 2.8.3.8, EC 3.1.2-, or EC 6.2.1.7,
an exogenous polynucleotide encoding a 3-HP-CoA dehydratase catalyzing conversion of 3-HP-CoA to acrylyl-CoA and belonging to EC 4.2.1,
an exogenous polynucleotide encoding a 3-HP-CoA hydrolase or a 3-hydroxyisobutyryl-CoA hydrolase belonging to EC 3.1.2,
a polynucleotide encoding an enzyme catalyzing the conversion of glycerol to 3-propionic aldehyde (3-HPA), and
a polynucleotide encoding an enzyme catalyzing the conversion of 3-HPA to 3-HP, and
wherein the genetically engineered microorganism has increased expression of the exogenous polynucleotides as compared to a microorganism of the same type that does not comprise the exogenous polynucleotides,
wherein the genetically engineered microorganism is an Escherichia , and
wherein the Escherichia produces 3-HP.

2. The microorganism of claim 1 , wherein the CoA transferase comprises an amino acid sequence selected from the group consisting of SEQ ID NOS: 1 to 10.

3. The microorganism of claim 1 , wherein the 3-HP-CoA dehydratase comprises an amino acid sequence selected from the group consisting of SEQ ID NOS: 21 to 98 and 401.

4. The microorganism of claim 1 , wherein the enzyme catalyzing conversion of acrylyl-CoA to acrylate comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 177 to 182.

5. The microorganism of claim 1 , wherein a gene that encodes at least one enzyme involved in a pathway of decomposing acrylate or converting acrylate to another product is deleted or disrupted in the microorganism.

6. The microorganism of claim 1 , wherein the genetically engineered microorganism is an Escherichia coli.

7. The microorganism of claim 6 , wherein the enzyme catalyzing conversion of glycerol to 3-HPA is glycerol dehydratase (dhaB) and the enzyme catalyzing conversion of 3-HPA to 3-HP is aldehyde dehydrogenase (AldH) or succinate semialdehyde dehydrogenase (gabD).

8. A method of producing acrylate, the method comprising culturing the microorganism of claim 1 in a culture medium to thereby produce acrylate.

9. The method of claim 8 , further comprising recovering the acrylate from the culture.

10. The microorganism of claim 1 , further comprising polynucleotides encoding a glycerol dehydratase reactivase (GDR).

11. A method of producing the genetically engineered microorganism according to claim 1 , the method comprising transforming an Escherichia microorganism with the exogenous polynucleotide encoding CoA transferase, the exogenous polynucleotide encoding 3-HP-CoA dehydratase, and the exogenous polynucleotide encoding the 3-HP-CoA hydrolase or the 3-hydroxyisobutyryl-CoA hydrolase, wherein the Escherichia microorganism comprises a polynucleotide encoding an enzyme catalyzing the conversion of glycerol to 3-propionic aldehyde (3-HPA), and a polynucleotide encoding an enzyme catalyzing the conversion of 3-HPA to 3-HP, and wherein the Escherichia produces 3-HP.

12. An expression vector comprising:
a polynucleotide encoding a coenzyme A (CoA) transferase catalyzing conversion of 3-hydroxypropionic acid (3-HP) to 3-hydroxypropionyl-CoA (3-HP-CoA) and belonging to EC 2.8.3.8, EC 3.1.2-, or EC 6.2.1.7,
a polynucleotide encoding a 3-HP-CoA dehydratase catalyzing conversion of 3-HP-CoA to acrylyl-CoA and belonging to EC 4.2.1,
a polynucleotide encoding a 3-HP-CoA hydrolase or a 3-hydroxyisobutyryl-CoA hydrolase belonging to EC 3.1.2, and
a polynucleotide that is heterologous to the polynucleotide encoding the CoA transferase, the polynucleotide encoding the 3-HP-CoA dehydratase, the polynucleotide encoding the 3-HP-CoA hydrolase, or the 3-hydroxyisobutyryl-CoA hydrolase.

13. The microorganism of claim 1 , wherein
the CoA transferase comprises an amino acid sequence selected from the group consisting of SEQ ID NOS: 1 to 10,
the 3-HP-CoA dehydratase comprises an amino acid sequence selected from the group consisting of SEQ ID NOS: 21 to 98 or 401, and
the 3 HP-CoA hydrolase or the 3-hydroxyisobutyryl-CoA hydrolase comprises an amino acid sequence selected from the group consisting of SEQ ID NOS: 177 to 182.

Assignees

• Samsung Electronics Co Ltd

Inventors

• Chu Hunsu
• Ahn Jinho
• Nam Taewook
• Yun Jiae
• Choi Insuk
• Song Yeoju
• Lee Jinsuk

Classifications

• C12Y301/02

  Thioester hydrolases (3.1.2) · CPC title

• C12Y602/01007

  Cholate--CoA ligase (6.2.1.7) · CPC title

• C12P7/40 Primary

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

• Y02P20/52

  using catalysts, e.g. selective catalysts · CPC title

• C12N9/16

  acting on ester bonds (3.1) · CPC title