Materials and methods for the biosynthesis of seven carbon chemicals in the presence of methanol oxidation
AssigneeInvista North America S.A.R.L.
InventorsAlex VAN ECK CONRADIE, Achuthanunni CHOKKATHUKALAM, Remi Ako Mbianyor MOMO
Life of this patent
Estimating the term…
Abstract
This disclosure describes methods for regulating the biosynthesis of pimelic acid, 7-aminoheptanoate, 7-hydroxyheptanoate, heptamethylenediamine, 7-aminoheptanol, or 1,7-heptanediol by channeling increased flux through the biosynthesis pathway to obtain an intermediate required for growth of the host microorganism.
Claims38
A method for regulating biosynthesis of a product chosen from pimelic acid, 7-aminoheptanoate, 7-hydroxyheptanoate, heptamethylenediamine, 7-aminoheptanol, and 1,7-heptanediol, or salts and derivatives thereof, using a pathway having a pimeloyl-ACP intermediate, the method comprising converting methanol to formate via at least one spontaneous enzymatic reaction, wherein the formate is used in the conversion of tetrahydrofolate to N10-formyl-tetrahydrofolate.
(canceled)
(canceled)
(canceled)
10-18. (canceled)
(canceled)
(canceled)
(canceled)
(canceled)
(canceled)
The method of claim 33, wherein the host is Escherichia coli.
(canceled)
(canceled)
(canceled)
A recombinant host for producing pimeloyl-ACP, wherein:
- the host comprises at least one exogenous nucleic acid encoding (i) a formate-tetrahydrofolate ligase, (ii) a S-(hydroxymethyl) glutathione dehydrogenase, and an (iii) a S-formylglutathione hydrolase;
- and
- the host optionally further comprises at least one exogenous nucleic acid encoding an alcohol dehydrogenase.
(canceled)
(canceled)
43-53. (canceled)
A nucleic acid construct or expression vector comprising:
- a polynucleotide encoding a polypeptide having formate-tetrahydrofolate ligase activity, wherein the polynucleotide is operably linked to one or more heterologous control sequences that direct production of the polypeptide and wherein the polypeptide having formate-tetrahydrofolate ligase activity is selected from:
- (a) a polypeptide having at least 70% sequence identity or homology to the polypeptide of SEQ ID NO:
- 18;
- (b) a polypeptide having at least 70% sequence identity or homology to the polypeptide of SEQ ID NO:
- 19;
- (c) a polypeptide having at least 70% sequence identity or homology to the polypeptide of SEQ ID NO:
- 20;
- (d) a polypeptide having at least 70% sequence identity or homology to the polypeptide of SEQ ID NO:
- 21;
- and (e) a polypeptide having at least 70% sequence identity or homology to the polypeptide of SEQ ID NO:
- 22;
- a polypeptide having alcohol dehydrogenase activity, wherein the polynucleotide is operably linked to one or more heterologous control sequences that direct production of the polypeptide and wherein the polypeptide having alcohol dehydrogenase is a polypeptide having at least 70% sequence identity or homology to the polypeptide of SEQ ID NO:
- 31;
- a polypeptide having S-(hydroxymethyl) glutathione dehydrogenase activity, wherein the polynucleotide is operably linked to one or more heterologous control sequences that direct production of the polypeptide and wherein the polypeptide having S-(hydroxymethyl) glutathione dehydrogenase activity is selected from:
- (a) a polypeptide having at least 70% sequence identity or homology to the polypeptide of SEQ ID NO:
- 23;
- (b) a polypeptide having at least 70% sequence identity or homology to the polypeptide of SEQ ID NO:
- 24;
- (c) a polypeptide having at least 70% sequence identity or homology to the polypeptide of SEQ ID NO:
- 25;
- and (d) a polypeptide having at least 70% sequence identity or homology to the polypeptide of SEQ ID NO:
- 26;
- or
- a polypeptide having S-formylglutathione hydrolase activity, wherein the polynucleotide is operably linked to one or more heterologous control sequences that direct production of the polypeptide and wherein the polypeptide having S-formylglutathione hydrolase activity is selected from:
- (a) a polypeptide having at least 70% sequence identity or homology to the polypeptide of SEQ ID NO:
- 27;
- (b) a polypeptide having at least 70% sequence identity or homology to the polypeptide of SEQ ID NO:
- 28;
- (c) a polypeptide having at least 70% sequence identity or homology to the polypeptide of SEQ ID NO:
- 29;
- and (d) a polypeptide having at least 70% sequence identity or homology to the polypeptide of SEQ ID NO:
- 30.
55-58. (canceled)
Dependencies are inferred from the claim text.