Enzymes and microorganisms for the production of 1,3-butadiene and other dienes

Patent metadata

Field	Value
Publication number	US-10125377-B2
Application number	US-201415022540-A
Country	US
Kind code	B2
Filing date	Aug 13, 2014
Priority date	Sep 18, 2013
Publication date	Nov 13, 2018
Grant date	Nov 13, 2018

Abstract

A method for the in vivo production of 1,3-butadiene from 2,4-pentadienoate is described (FIG. 1 ). Enzymes capable of decarboxylating 2,4-pentadienoate to 1,3-butadiene have been discovered. Recombinant expression of these newly discovered enzymes has resulted in the engineering of microorganisms capable of producing 1,3-butadiene when cultured in the presence of 2,4-pentadienoate. 1,3-butadienoate is an important monomer used in the manufacturing of rubbers and plastics. This invention will help to enable the biological production of 1,3-butadiene from renewable resources such as sugar, for example.

First claim

What is claimed is:

1. A method for production of 1,3-butadiene from a recombinant cell comprising:
(i) contacting a recombinant host cell with at least one carbon source, the recombinant host cell including the ability to:
over produce 2,4-pentadienoate; and
express a gene with SEQ ID NO: 1 encoding a polypeptide having 2,4-pentadienoate decarboxylase activity; and

(ii) culturing said recombinant cell under conditions sufficient to produce 1,3-butadiene, wherein expression of the gene with SEQ ID NO: 1 encoding the polypeptide having 2,4-pentadienoate decarboxylase activity provides increased production of 1,3-butadiene in the recombinant host cell relative to a control recombinant host cell lacking expression of the gene with SEQ ID NO: 1 encoding the polypeptide having 2,4-pentadienoate decarboxylase activity.

2. The method according to claim 1 wherein the polypeptide having 2,4-pentadienoate decarboxylase activity has trans-2,4-pentadienoate decarboxylase activity.

3. The method according to claim 1 wherein the polypeptide having 2,4-pentadienoate decarboxylase activity has cis-2,4-pentadienoate decarboxylase activity.

4. The method according to claim 1 wherein the gene with SEQ ID NO: 1 encoding the polypeptide having 2,4-pentadienoate decarboxylase activity is derived from Saccharomyces cerevisiae.

5. The method according to claim 1 wherein the gene with SEQ ID NO: 1 encoding the polypeptide having 2,4-pentadienoate decarboxylase activity comprises fdc1 from Saccharomyces cerevisiae (SEQ ID NO: 1).

6. The method according to claim 1 wherein the gene with SEQ ID NO: 1 encoding the polypeptide having 2,4-pentadienoate decarboxylase activity is derived from an organism that comprises at least one member selected from the group consisting of bacteria, yeast, filamentous fungi, cyanobacteria, algae, and plant cells.

7. The method according to claim 1 wherein the gene with SEQ ID NO: 1 encoding the polypeptide having 2,4-pentadienoate decarboxylase activity is derived from an organism that comprises at least one member selected from the group consisting of Escherichia, Salmonella, Bacillus, Acinetobacter, Streptomyces, Corynebacterium, Methylosinus, Methylomonas, Rhodococcus, Pseudomonas, Rhodobacter, Synechocystis, Saccharomyces, Klebsiella, Zygosaccharomyces, Kluyveromyces, Candida, Hansuela, Debaryomyces, Mucor, Pichia, Torulopsis, Aspergillus, Arthrobotrys, Brevibacterium, Microbacterium, Arthrobacter, Citrobacter, Chlamydomonas, Fusarium, Penicillium , and Zymomonas.

8. The method according to claim 1 wherein the recombinant host cell comprises at least one member selected from the group consisting of bacteria, yeast, filamentous fungi, cyanobacteria, algae, and plant cells.

9. The method according to claim 1 wherein the recombinant host cell comprises at least one member selected from the group consisting of Escherichia, Salmonella, Bacillus, Acinetobacter, Streptomyces, Corynebacterium, Cupriavidus, Clostridium, Methylosinus, Methylomonas, Rhodococcus, Pseudomonas, Rhodobacter, Synechocystis, Saccharomyces, Klebsiella, Zygosaccharomyces, Kluyveromyces, Candida, Hansuela, Debaryomyces, Mucor, Pichia, Torulopsis, Aspergillus, Arthrobotrys, Brevibacterium, Microbacterium, Arthrobacter, Citrobacter, Chlamydomonas, Fusarium, Penicillium , and Zymomonas.

10. The method according to claim 1 wherein the recombinant host cell includes the natural ability to over produce 2,4-pentadienoate.

11. The method according to claim 1 wherein the recombinant host cell further comprises a recombinant host cell previously engineered to over produce 2,4-pentadienoate.

12. The method according to claim 1 wherein the recombinant host cell comprises a strain that overproduces at least one member selected from the group consisting of aromatic amino acids, chorismate, isochorismate, dihydroshikimate, protocatechuate, salicylate, p-hydroxybenzoate, phenol, catechol, muconate, and 2,4-pentadienoate.

13. The method according to claim 1 wherein the at least one carbon source comprises at least one member selected from the group consisting of monosaccharides, oligosaccharides, polysaccharides, glycerol, carbon dioxide, carbon monoxide, methanol, formaldehyde, formate, amino acids, and carbon-containing amines.

Assignees

• Nielsen David
• Pugh Shawn
• Mckenna Rebekah
• Univ Arizona State

Inventors

• Nielsen David
• Pugh Shawn
• Mckenna Rebekah

Classifications

• C12P5/026 Primary

  Unsaturated compounds, i.e. alkenes, alkynes or allenes · CPC title

• Y02E50/343

  Cross-Sectional Technologies · mapped topic

• C12N9/88

  Lyases (4.) · CPC title

• C12Y401/01

  Carboxy-lyases (4.1.1) · CPC title

• Y02E50/30

  Fuel from waste, e.g. synthetic alcohol or diesel · CPC title