Materials and Methods of Producing 7-Carbon Monomers
AssigneeInvista North America S.A.R.L.
InventorsAdriana Leonora Botes, Alex Van Eck Conradie
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Abstract
This document describes materials and methods for producing 7-hydroxyheptanoic acid using a β-ketothiolase or a synthase and an alcohol O-acetyltransferase to form a 7-acetyloxy-3-oxoheptanoyl-CoA intermediate. This document describes biochemical pathways for producing 7-hydroxyheptanoic acid using a β-ketothiolase or a synthase and an alcohol O-acetyltransferase to form a 7-acetyloxy-3-oxoheptanoyl-CoA intermediate. 7-hydroxyheptanoic acid can be enzymatically converted to pimelic acid, 7-aminoheptanoic acid, heptamethylenediamine or 1,7 heptanediol. This document also describes recombinant hosts producing 7-hydroxyheptanoic acid as well as pimelic acid, 7-aminoheptanoic acid, heptamethylenediamine and 1,7 heptanediol.
Claims56
A method of producing 7-acetyloxy-3-oxoheptanoyl-CoA, said method comprising enzymatically converting 5-acetyloxypentanoyl-CoA to 7-acetyloxy-3-oxoheptanoyl-CoA using a polypeptide having the activity of a β-ketothiolase or synthase classified under EC. 2.3.1.-.
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-11. (canceled)
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A method for biosynthesizing 7-hydroxyheptanoate, said method comprising enzymatically synthesizing 7-acetyloxy-3-oxoheptanoyl-CoA from 5-acetyloxypentanoyl-CoA using a polypeptide having the activity of a β-ketothiolase or synthase classified under EC. 2.3.1.- and enzymatically converting 7-acetyloxy-3-oxoheptanoyl-CoA to 7-hydroxyheptanoate.
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-22. (canceled)
-25. (canceled)
-32. (canceled)
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-44. (canceled)
A non-naturally occurring recombinant host comprising at least one exogenous nucleic acid encoding (i) an alcohol O-acetyltransferase, (ii) a β-ketothiolase or synthase, (iii) a thioesterase or a CoA transferase, (v) an esterase and one or more of (vi) a 3-hydroxyacyl-CoA dehydrogenase or a 3-oxoacyl-CoA reductase, (vii) an enoyl-CoA hydratase, and (viii) a trans-2-enoyl-CoA reductase, said host producing 7-hydroxyheptanoate.
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-56. (canceled)
A biochemical network comprising a β-ketothiolase or synthase classified under EC. 2.3.1.-, 5-acetyloxypentanoyl-CoA, and 7-acetyloxy-3-oxoheptanoyl-CoA, wherein said β-ketothiolase or synthase enzymatically converts 5-acetyloxypentanoyl-CoA to 7-acetyloxy-3-oxoheptanoyl-CoA.
A means for producing 7-acetyloxy-3-oxoheptanoyl-CoA, wherein said means enzymatically converts 5-acetyloxypentanoyl-CoA to 7-acetyloxy-3-oxoheptanoyl-CoA.
A step for obtaining 7-acetyloxy-3-oxoheptanoyl-CoA using a β-ketothiolase or synthase classified under EC. 2.3.1.-.
A composition comprising a bio-based, bio-derived or fermentation-derived 5-acetyloxypentanoyl-CoA, bio-based, bio-derived or fermentation-derived 7-acetyloxy-3-oxoheptanoyl-CoA, and a β-ketothiolase or synthase classified under EC. 2.3.1.-.
A composition comprising bio-based, bio-derived or fermentation-derived 7-acetyloxy-3-oxoheptanoyl-CoA.
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A bio-based, bio-derived or fermentation-derived 7-acetyloxy-3-oxoheptanoyl-CoA produced by the method of enzymatically converting 5-hydroxypentanoate to 7-acetyloxy-3-oxoheptanoyl-CoA in one or more enzymatic steps using (i) a β-ketothiolase or synthase and (ii) and an alcohol O-acetyltransferase classified under EC. 2.3.1.-.
A non-naturally occurring organism comprising at least one exogenous nucleic acid encoding at least one polypeptide having the activity of at least one enzyme depicted in any one of FIGS. 1 to 5.
A nucleic acid construct or expression vector comprising
- (a) a polynucleotide encoding a polypeptide having the activity of a β-ketothiolase, wherein the polynucleotide is operably linked to one or more heterologous control sequences that direct production of the polypeptide and wherein the polypeptide having the activity of a β-ketothiolase is selected from the group consisting of a polypeptide having at least 70% sequence identity to the polypeptide of SEQ ID NO:
- 1 or SEQ ID NO:
- 16;
- (b) a polynucleotide encoding a polypeptide having the activity of a trans-2-enoyl-CoA reductase, wherein the polynucleotide is operably linked to one or more heterologous control sequences that direct production of the polypeptide and wherein the polypeptide having the activity of a trans-2-enoyl-CoA reductase is selected from the group consisting of a polypeptide having at least 70% sequence identity to the polypeptide of SEQ ID NO:
- 23, SEQ ID NO:
- 24, SEQ ID NO:
- 25, SEQ ID NO:
- 26, SEQ ID NO:
- 27 or SEQ ID NO:
- 28;
- (c) a polynucleotide encoding a polypeptide having the activity of ω-transaminase, wherein the polynucleotide is operably linked to one or more heterologous control sequences that direct production of the polypeptide and wherein the polypeptide having the activity of ω-transaminase is selected from the group consisting of a polypeptide having at least 70% sequence identity to the polypeptide of SEQ ID NO:
- 7, SEQ ID NO:
- 8, SEQ ID NO:
- 9, SEQ ID NO:
- 10, SEQ ID NO:
- 11, or SEQ ID NO:
- 12;
- (d) a polynucleotide encoding a polypeptide having the activity of a phosphopantetheinyl transferase, wherein the polynucleotide is operably linked to one or more heterologous control sequences that direct production of the polypeptide and wherein the polypeptide having phosphopantetheinyl transferase is selected from the group consisting of a polypeptide having at least 70% sequence identity to the polypeptide of SEQ ID NO:
- 13 or 14;
- (e) a polynucleotide encoding a polypeptide having the activity of an alcohol-O-acetyltransferase, wherein the polynucleotide is operably linked to one or more heterologous control sequences that direct production of the polypeptide and wherein the polypeptide having the activity of an alcohol-O-acetyltransferase is selected from the group consisting of a polypeptide having at least 70% sequence identity to the polypeptide of SEQ ID NO:
- 17;
- (f) a polynucleotide encoding a polypeptide having the activity of a carboxylate reductase, wherein the polynucleotide is operably linked to one or more heterologous control sequences that direct production of the polypeptide and wherein the polypeptide having the activity of a carboxylate reductase is selected from the group consisting of a polypeptide having at least 70% sequence identity to the polypeptide of SEQ ID NO:
- 2, SEQ ID NO:
- 3, SEQ ID NO:
- 4, SEQ ID NO:
- 5, SEQ ID NO:
- 6 or SEQ ID NO:
- 29;
- (g) a polynucleotide encoding a polypeptide having the activity of a esterase, wherein the polynucleotide is operably linked to one or more heterologous control sequences that direct production of the polypeptide and wherein the polypeptide having the activity of a esterase is selected from the group consisting of a polypeptide having at least 70% sequence identity to the polypeptide of SEQ ID NO:
- 15;
- (h) a polynucleotide encoding a polypeptide having the activity of a pimeloyl-[acp] methyl ester esterase, wherein the polynucleotide is operably linked to one or more heterologous control sequences that direct production of the polypeptide and wherein the polypeptide having the activity of a pimeloyl-[acp] methyl ester esterase is selected from the group consisting of a polypeptide having at least 70% sequence identity to the polypeptide of SEQ ID NO:
- 18;
- (i) a polynucleotide encoding a polypeptide having the activity of a CoA-transferase, wherein the polynucleotide is operably linked to one or more heterologous control sequences that direct production of the polypeptide and wherein the polypeptide having the activity of a CoA-transferase is selected from the group consisting of a polypeptide having at least 70% sequence identity to the polypeptide of SEQ ID NO:
- 19;
- or
- (j) a polynucleotide encoding a polypeptide having the activity of a decarboxylase, wherein the polynucleotide is operably linked to one or more heterologous control sequences that direct production of the polypeptide and wherein the polypeptide having the activity of a decarboxylase is selected from the group consisting of a polypeptide having at least 70% sequence identity to the polypeptide of SEQ ID NO:
- 20, SEQ ID NO:
- 21, or SEQ ID NO:
- 22.
Dependencies are inferred from the claim text.