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US 7,582,457 B2Granted
Method for the production of 1,3-propanediol by recombinant organisms comprising genes for coenzyme B12 synthesis
AssigneesE.I. du Pont de Nemours and Company, Genencor International, Inc.
InventorsNigel S. Dunn-Coleman, Anthony Gatenby, Fernando Valle
Life of this patent
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Abstract
Recombinant organisms are provided comprising genes encoding cob(II)alamin reductase, cob(I)alamin adenosyltransferase, glycerol dehydratase and 1,3-propanediol oxidoreductase activities useful for the production of 1,3-propanediol from a variety of carbon substrates.
Claims9
A process for the bio-production of 1,3-propanediol comprising:
- (i) contacting a transformed host cell with at least one fermentable carbon source and an effective amount of coenzyme B12 precursor whereby 1,3-propanediol is produced, the transformed host cell comprising:
- (a) at least one copy of genes encoding a protein having a glycerol dehydratase activity or a diol dehydratase activity selected from the group consisting of Klebsiella pneumoniae dhaB1-3 as shown in SEQ ID NO:4, SEQ ID NO:5, and SEQ ID NO:6;
- (b) at least one copy of a gene encoding a protein having a 1,3-propanediol oxidoreductase activity of Klebsiella pneumoniae dhaT as shown in SEQ ID NO:7;
- and
- (c) one copy of a gene encoding a protein having a cob(I)alamin adenosyltransferase activity selected from the group consisting of Escherichia coil btuR as shown in SEQ ID NO:1, Salmonella typhimurium cobA as shown in SEQ ID NO:2, and Pseudomonas denitrificans cobO as shown in SEQ ID NO:3;
- wherein the genes of (i)(a)-(i)(c) are heterologous to the host cell and express their respective gene products in active form, and
- (ii) recovering the 1,3-propanediol produced from step (i).
4 more elements
An improved process for the bio-production of 1,3-propanediol comprising:
- (i) contacting a transformed host cell with at least one carbon source selected from the group consisting of fermentable carbohydrates, single-carbon substrates and mixtures thereof, and an effective amount of coenzyme B12 precursor, whereby 1,3-propanediol is produced;
- the transformed host cell comprising:
- (a) at least one copy of genes encoding a protein having a glycerol dehydratase activity or a diol dehydratase activity selected from the group consisting of Klebsiella pneumoniae dhaB1-3 as shown in SEQ ID NO:4, SEQ ID NO:5, and SEQ ID NO:6;
- (b) at least two copies of a gene encoding a protein having a 1,3-propanediol oxidoreductase activity of Klebsiella pneumoniae dhaT as shown in SEQ ID NO:7;
- and
- (c) at least one copy of a gene encoding a protein having a cob(I)alamin adenosyltransferase activity selected from the group consisting of Escherichia coil btuR as shown in SEQ ID NO:1, Salmonella typhimurium cobA as shown in SEQ ID NO:2, and Pseudomonas denitrificans cobO as shown in SEQ ID NO:3;
- wherein the genes of (i)(a)-(i)(c) are heterologous and express their respective gene products in active form;
- and
- (ii) recovering the 1,3-propanediol produced from step (i);
- the improvement comprising an increase in the production of 1,3-propanediol as compared with a bio-process wherein the gene of (i)(c) is not present in multicopy in the transformed host cell.
7 more elements
A process for regulating the bio-production of 1,3-propanediol comprising:
- (i) contacting a transformed host cell with (a) at least one carbon source selected from the group consisting of fermentable carbohydrates, single-carbon substrates and mixtures thereof and (b) an effective amount of coenzyme B12 precursor whereby 1,3-propanediol is produced, the transformed host cell comprising:
- (a)at least one copy of genes encoding a protein having a glycerol dehydratase activity or a diol dehydratase activity selected from the group consisting of Klebsiella pneumoniae dhaB1-3 as shown in SEQ ID NO:4, SEQ ID NO:5, and SEQ ID NO:6;
- (b) at least one copy of a gene encoding a protein having a 1,3-propanediol oxidoreductase activity of Klebsiella pneumoniae dhaT as shown in SEQ ID NO:7;
- and
- (c) at least one copy of a gene encoding a protein having a cob(I)alamin adenosyltransferase activity selected from the group consisting of Escherichia coil btuR as shown in SEQ ID NO:1, Salmonella typhimurium cobA as shown in SEQ ID NO:2, and Pseudomonas denitrificans cobO as shown in SEQ ID NO:3;
- wherein the gene of (i)(c) is selectively inhibited whereby the metabolism of coenzyme B12 precursor is regulated.
3 more elements
Dependencies are inferred from the claim text.