WO1998021341B1 - Method for the recombinant production of 1,3-propanediol - Google Patents

Method for the recombinant production of 1,3-propanediol

Info

Publication number
WO1998021341B1
WO1998021341B1 PCT/US1997/020873 US9720873W WO9821341B1 WO 1998021341 B1 WO1998021341 B1 WO 1998021341B1 US 9720873 W US9720873 W US 9720873W WO 9821341 B1 WO9821341 B1 WO 9821341B1
Authority
WO
WIPO (PCT)
Prior art keywords
protein
nucleic acid
acid encoding
microorganism
seq
Prior art date
Application number
PCT/US1997/020873
Other languages
French (fr)
Other versions
WO1998021341A3 (en
WO1998021341A2 (en
Filing date
Publication date
Application filed filed Critical
Priority to BRPI9714313-8A priority Critical patent/BR9714313B1/en
Priority to NZ335392A priority patent/NZ335392A/en
Priority to AU55076/98A priority patent/AU735080B2/en
Priority to DE69736070T priority patent/DE69736070T2/en
Priority to JP52287098A priority patent/JP2001503636A/en
Priority to DK97951433T priority patent/DK0946732T3/en
Priority to US09/308,207 priority patent/US6953684B2/en
Priority to CA002270906A priority patent/CA2270906A1/en
Priority to EP97951433A priority patent/EP0946732B1/en
Publication of WO1998021341A2 publication Critical patent/WO1998021341A2/en
Publication of WO1998021341A3 publication Critical patent/WO1998021341A3/en
Publication of WO1998021341B1 publication Critical patent/WO1998021341B1/en

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Abstract

The present invention provides an improved method for the production of 1,3-propanediol from a variety of carbon sources is an organism comprising DNA encoding protein X of a dehydratase or protein X in combination with at least one of protein 1, protein 2 and protein 3. The protein X may be isolated from a diol dehydratase or a glycerol dehydratase. The present invention also provides host cells comprising protein X that are capable of increased production of 1,3-propanediol.

Claims

AMENDED CLAIMS[received by the International Bureau on 17 June 1998 (17.06.98); original claims 1-40 replaced by amended claims 1-40 (4 pages)]
1. An improved method for the production of 1 ,3-propanediol from a microorganism comprising the steps of: a) obtaining a recombinant microorganism capable of producing 1 ,3- propanediol, said microorganism comprising at least one nucleic acid encoding a dehydratase activity and a nucleic acid encoding protein X; and b) culturing the recombinant microorganism in the presence of at least one carbon source capable of being converted to 1 ,3 propanediol in said transformed microorganism and under conditions suitable for the production of 1 ,3 propanediol wherein the carbon source is selected from the group consisting of monosaccharides, oligosaccharides, polysaccharides, and a one carbon substrate.
2. The method of Claim 1 wherein said recombinant microorganism comprises at least one nucleic acid encoding a protein selected from the group consisting of protein 1 , protein 2 and protein 3.
3. The method of Claim 1 further comprising the step of recovering the 1 ,3 propanediol.
4. The method of Claim 1 wherein the nucleic acid encoding protein X is isolated from a glycerol dehydratase gene cluster.
5. The method of Claim 1 wherein the nucleic acid encoding protein X is isolated from a diol dehydratase gene cluster.
6. The method of Claim 4 wherein the glycerol dehydratase gene cluster is from an organism selected from the genera consisting of Kiebsiella and Citrobactor.
7. The method of Claim 5 wherein the diol dehydratase gene cluster is from an organism selected from the genera consisting of Kiebsiella, Clostridium and Salmonella.
8. The method of Claim 1 wherein the nucleic acid encoding a dehydratase activity is heterologous to the organism.
9. The method of Claim 1 wherein the nucleic acid encoding a dehydratase activity is homologous to the organism. ΓÇö 106 ΓÇö
10. The method of Claim 1 wherein the recombinant microorganism is selected from the group of genera consisting of Citrobacter, Enterobacter, Clostridium, Kiebsiella, Aerobacter, Lactobacillus, Aspergillus, Saccharomyces, Schizosaccharomyces, Zygo saccharomyces, Pichia, Kluyveromyces, Candida, Hansenula, Debaryomyces, Mucor, Torulopsis, Methylobacter, Escherichia, Salmonella, Bacillus, Streptomyces and Pseudomonas.
11. The method of Claim 10 wherein the microorganism is selected from the group consisting of E.coli and Kiebsiella spp.
12. The method of Claim 1 wherein the nucleic acid encoding protein X is stably maintained in the host genome.
13. The method of Claim 2 wherein at least one nucleic acid encoding a protein selected from protein 1 , protein 2 and protein 3 is stably maintained in the host genome.
14. The method of Claim 1 wherein the carbon source is glucose.
15. The method of Claim 1 wherein the nucleic acid encoding protein X has the sequence as shown in SEQ ID NO: 59.
16. The method of Claim 2 wherein protein 1 has the sequence as shown in SEQ ID NO: 60 or SEQ ID NO: 61.
17. The method of Claim 2 wherein protein 2 has the sequence as shown in SEQ ID NO: 62 or SEQ ID NO: 63.
18. The method of Claim 2 wherein protein 3 has the sequence as shown in SEQ ID NO:64 or SEQ ID NO: 65.
19. A recombinant microorganism capable of producing 1 ,3-propanediol from a carbon source said recombinant microorganism comprising a) at least one nucleic acid encoding a dehydratase activity; b) at least one nucleic acid encoding a glycerol-3-phosphatase; and c) at least one nucleic acid encoding protein X.
20. The recombinant microorganism of Claim 19 further comprising d) at least one nucleic acid encoding a protein selected from the group consisting of protein 1 , protein 2 and protein 3. ΓÇö 107 ΓÇö
21. The recombinant microorganism of Claim 19 selected from the group consisting of Citrobacter, Enterobacter, Clostridium, Kiebsiella, Aerobacter, Lactobacillus, Aspergillus, Saccharomyces, Schizosaccharomyces, Zygosaccharomyces, Pichia, Kluyveromyces, Candida, Hansenula, Debaryomyces, Mucor, Torulopsis, Methylobacter, Escherichia, Salmonella, Bacillus, Streptomyces and Pseudomonas.
22. The recombinant microorganism of Claim 19 wherein the nucleic acid encoding protein X is isolated from a glycerol dehydratase gene cluster.
23. The recombinant microorganism of Claim 19 wherein the nucleic acid encoding protein X is isolated from a diol dehydratase gene cluster.
24. The recombinant microorganism of Claim 22 wherein the glycerol dehydratase gene cluster is from an organism selected from the genera consisting of Kiebsiella and Citrobactor.
25. The recombinant microorganism of Claim 23 wherein the diol dehydratase gene cluster is from an organism selected from the genera consisting of Kiebsiella, Clostridium and Salmonella.
26. The recombinant microorganism of Claim 19 wherein said dehydratase activity is heterologous to said microorganism.
27. The recombinant microorganism of Claim 19 wherein said dehydratase activity is homologous to said microorganism.
28. The recombinant microorganism of Claim 19 wherein the nucleic acid encoding protein X has the sequence as shown in SEQ ID NO: 59.
29. The recombinant microorganism of Claim 20 wherein protein 1 has the sequence as shown in SEQ ID NO: 60 or SEQ ID NO: 61.
30. The recombinant microorganism of Claim 20 wherein protein 2 has the sequence as shown in SEQ ID NO: 62 or SEQ ID NO: 63. ΓÇö 108 ΓÇö
31. The recombinant of Claim 20 wherein protein 3 has the sequence as shown in SEQ ID: 64 or SEQ ID NO: 65.
32. A method for extending the half-life of dehydratase activity in a transformed microorganism capable of producing 1 ,3-propanediol and containing at least one nucleic acid encoding a dehydratase activity, comprising the step of introducing a nucleic acid encoding protein X into said microorganism and culturing under conditions suitable for production of 1 ,3- propanediol.
33. The method of Claim 32 wherein the nucleic acid encoding the dehydratase activity is heterologous to said microorganism.
34. The method of Claim 32 wherein the nucleic acid encoding the dehydratase activity is homologous to said microorganism.
35. The method of Claim 32 wherein the nucleic acid encoding protein X is isolated from a glycerol dehydratase gene cluster.
36. The method of Claim 32 wherein the nucleic acid encoding protein X is isolated from a diol dehydratase gene cluster.
37. The method of Claim 35 wherein the glycerol dehydratase gene cluster is from an organism selected from the genera consisting of Kiebsiella and Citrobactor.
38. The method of Claim 34 wherein the diol dehydratase gene cluster is from an organism selected from the genera consisting of Kiebsiella, Clostridium and Salmonella.
39. The method of Claim 32 wherein the microorganism is selected from the group consisting of Citrobacter, Enterobacter, Clostridium, Kiebsiella, Aerobacter, Lactobacillus, Aspergillus, Saccharomyces, Schizosaccharomyces, Zygosaccharomyces, Pichia, Kluyveromyces, Candida, Hansenula, Debaryomyces, Mucor, Torulopsis, Methylobacter, Escherichia, Salmonella, Bacillus, Streptomyces and Pseudomonas.
40. The method of Claim 32 further comprising the step of introducing at least one nucleic acid encoding protein 1 , protein 2 or protein 3 into said microorganism.
PCT/US1997/020873 1996-11-13 1997-11-13 Method for the recombinant production of 1,3-propanediol WO1998021341A2 (en)

Priority Applications (9)

Application Number Priority Date Filing Date Title
BRPI9714313-8A BR9714313B1 (en) 1996-11-13 1997-11-13 METHOD FOR THE PRODUCTION OF 1,3-PROPANODIOL RECOMBINANTS
NZ335392A NZ335392A (en) 1996-11-13 1997-11-13 Method for the recombinant production of 1,3-propanediol
AU55076/98A AU735080B2 (en) 1996-11-13 1997-11-13 Method for the recombinant production of 1,3-propanediol
DE69736070T DE69736070T2 (en) 1996-11-13 1997-11-13 Recombinant method of preparation of 1,3-propanediol
JP52287098A JP2001503636A (en) 1996-11-13 1997-11-13 Method for recombinant production of 1,3-propanediol
DK97951433T DK0946732T3 (en) 1996-11-13 1997-11-13 Process for the Recombinant Preparation of 1,3-Propanediol
US09/308,207 US6953684B2 (en) 1996-11-13 1997-11-13 Method for the recombinant production of 1,3-propanediol
CA002270906A CA2270906A1 (en) 1996-11-13 1997-11-13 Method for the recombinant production of 1,3-propanediol
EP97951433A EP0946732B1 (en) 1996-11-13 1997-11-13 Method for the recombinant production of 1,3-propanediol

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US3060196P 1996-11-13 1996-11-13
US60/030,601 1996-11-13

Publications (3)

Publication Number Publication Date
WO1998021341A2 WO1998021341A2 (en) 1998-05-22
WO1998021341A3 WO1998021341A3 (en) 1998-06-25
WO1998021341B1 true WO1998021341B1 (en) 1998-07-30

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PCT/US1997/020292 WO1998021339A1 (en) 1996-11-13 1997-11-10 Method for the production of 1,3-propanediol by recombinant organisms
PCT/US1997/020873 WO1998021341A2 (en) 1996-11-13 1997-11-13 Method for the recombinant production of 1,3-propanediol

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US (3) US6013494A (en)
EP (2) EP0942989B1 (en)
JP (2) JP4327909B2 (en)
KR (1) KR100509310B1 (en)
CN (2) CN1236063C (en)
AT (2) ATE452979T1 (en)
AU (2) AU733534B2 (en)
BR (2) BR9712761B1 (en)
CA (2) CA2269088C (en)
DE (2) DE69739715D1 (en)
DK (1) DK0946732T3 (en)
ES (1) ES2336858T3 (en)
ID (1) ID21487A (en)
IL (2) IL129722A0 (en)
MY (1) MY131754A (en)
NZ (1) NZ335392A (en)
WO (2) WO1998021339A1 (en)
ZA (1) ZA9710194B (en)

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