Enzymes and Microorganisms for the Production of 1,3-Butadiene and Other Dienes

Patent metadata

Field	Value
Publication number	US-2016230194-A1
Application number	US-201415022540-A
Country	US
Kind code	A1
Filing date	Aug 13, 2014
Priority date	Sep 18, 2013
Publication date	Aug 11, 2016
Grant date	—

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 the production of 1,3-butadiene from a recombinant cell comprising:
culturing a recombinant host cell with 2,4-pentadienoate, the recombinant host cell expressing at least one gene encoding at least one polypeptide configured for 2,4-pentadienoate activity, wherein expression in the recombinant host cell of the at least one gene encoding at least one polypeptide configured for 2,4-pentadienoate activity provides increased production of 1,3-butadiene in the recombinant host cell relative to a control recombinant host cell lacking expression of the at least one gene encoding the at least one polypeptide configured for 2,4-pentadienoate activity.

2 . A method according to claim 1 wherein the at least one polypeptide configured for 2,4-pentadienoate activity is configured for trans-2,4-pentadienoate decarboxylase activity.

3 . A method according to claim 1 wherein the at least one polypeptide configured for 2,4-pentadienoate activity is configured for cis-2,4-pentadienoate decarboxylase activity.

4 . A method according to claim 1 wherein a sequence of the at least one gene encoding the at least one polypeptide configured for 2,4-pentadienoate activity consists of SEQ ID NO:1.

5 . A method according to claim 1 wherein the at least one gene encoding the at least one polypeptide configured for 2,4-pentadienoate activity is derived from saccharomyces cerevisiae.

6 . A method according to claim 1 wherein the at least one gene encoding the at least one polypeptide configured for 2,4-pentadienoate activity comprises fdc1 from saccharomyces cerevisiae (SEQ ID NO: 1).

7 . A method according to claim 1 wherein the at least one gene encoding the at least one polypeptide configured for 2,4-pentadienoate 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.

8 . A method according to claim 1 wherein the at least one gene encoding the at least one polypeptide configured for 2,4-pentadienoate 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, hansenula, debaryomyces, mucor, pichia, torulopsis, aspergillus, arthrobotrys, brevibacterium, microbacterium, arthrobacter, ctirobacter, chlamydomonas, fusarium, penicillium , and zymomonas.

9 . A 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.

10 . A 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, hansenula, debaryomyces, mucor, pichia, torulopsis, aspergillus, arthrobotrys, brevibacterium, microbacterium, arthrobacter, ctirobacter, chlamydomonas, fusarium, penicillium , and zymomonas.

11 . A method for the 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 at least one gene encoding at least one 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 at least one gene encoding at least one 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 at least one gene encoding the at least one polypeptide having 2,4-pentadienoate decarboxylase activity.

12 . A method according to claim 11 wherein the at least one polypeptide having 2,4-pentadienoate decarboxylase activity is configured for trans-2,4-pentadienoate decarboxylase activity.

13 . A method according to claim 11 wherein the at least one polypeptide having 2,4-pentadienoate decarboxylase activity is configured for cis-2,4-pentadienoate decarboxylase activity.

14 . A method according to claim 11 wherein the sequence of the at least one gene encoding the at least one polypeptide having 2,4-pentadienoate decarboxylase activity comprises seq id no:1.

15 . A method according to claim 11 wherein the at least one gene encoding the at least one polypeptide configured for 2,4-pentadienoate activity is derived from saccharomyces cerevisiae.

16 . A method according to claim 11 wherein the at least one gene encoding the at least one polypeptide configured for 2,4-pentadienoate activity comprises fdc1 from saccharomyces cerevisiae (SEQ ID NO: 1).

17 . A method according to claim 11 wherein the at least one gene encoding the at least one polypeptide configured for 2,4-pentadienoate 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.

18 . A method according to claim 11 wherein the at least one gene encoding the at least one polypeptide configured for 2,4-pentadienoate 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, hansenula, debaryomyces, mucor, pichia, torulopsis, aspergillus, arthrobotrys, brevibacterium, microbacterium, arthrobacter, ctirobacter, chlamydomonas, fusarium, penicillium , and zymomonas.

19 . A method according to claim 11 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.

20 . A method according to claim 11 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, hansenula, debaryomyces, mucor, pichia, torulopsis, aspergillus, arthrobotrys, brevibacterium, microbacterium, arthrobacter, ctirobacter, chlamydomonas, fusarium, penicillium , and zymomonas.

21 . A method according to claim 11 wherein the recombinant host cell includes the natural ability to over produce 2,4-pentadienoate.

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

23 . A method according to claim 11 wherein the recombinant host cell comprises a strain that overproduces at least one m

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

• 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