Claims
- 1. A method of increasing production of methionine by a microorganism comprising culturing a microorganism which overexpresses a positive regulator of methionine biosynthesis under conditions such that production of methionine is increased.
- 2. The method of claim 1, wherein said positive regulator of methionine biosynthesis is RXN02910.
- 3. The method of claim 2, wherein RXN02910 is encoded by a nucleic acid molecule comprising the nucleotide sequence set forth as SEQ ID NO: 5 or SEQ ID NO: 16.
- 4. The method of claim 1, wherein said microorganism has been transformed with a plasmid comprising the nucleotide sequence of SEQ ID NO: 13.
- 5. The method of claim 1, wherein said positive regulator of methionine biosynthesis is a transcriptional regulatory protein.
- 6. A method of increasing production of methionine by a microorganism comprising culturing a microorganism which is deficient in a negative regulator of methionine biosynthesis or a microorganism which exhibits a decreased activity of a negative regulator of methionine biosynthesis under conditions such that production of methionine is increased.
- 7. The method of claim 6, wherein said negative regulator of methionine biosynthesis is RXA00655.
- 8. The method of claim 7, wherein RXA00655 is encoded by a nucleic acid molecule comprising the nucleotide sequence set forth as SEQ ID NO: 1, SEQ ID NO: 18, SEQ ID NO: 20, or SEQ ID NO: 22.
- 9. The method of claim 6, wherein said microorganism has been transformed with a plasmid comprising the nucleotide sequence of SEQ ID NO: 3.
- 10. The method of claim 6, wherein said negative regulator of methionine biosynthesis is a transcriptional regulatory protein.
- 11. The method of claim 6, wherein deficiency or decreased activity is achieved by a method selected from the group consisting of:
a) knock-out of the gene encoding said negative regulatory protein; b) mutagenesis of the gene encoding said negative regulatory protein, wherein said mutation can be induced in the coding, non-coding, or regulatory regions of said gene; c) expression of an anti-sense RNA, wherein said anti-sense RNA is complementary to at least part of the RNA encoding said negative regulatory protein; d) expression of DNA-binding proteins blocking or reducing expression from the gene encoding said negative regulatory protein; e) expression of protein-binding factors blocking or reducing expression from the gene encoding said negative regulatory protein; f) expression of a dominant-negative variant of said negative regulatory protein; and g) destabilization of the mRNA encoding said negative regulatory protein.
- 12. A method of producing methionine comprising culturing a microorganism which overexpresses RXN02910 under conditions such that methionine is produced.
- 13. A method of producing methionine comprising culturing a microorganism which has suppressed expression of RXA00655 under conditions such that methionine is produced.
- 14. A method of producing methionine comprising culturing a microorganism which has increased RXN02910 activity under conditions such that methionine is produced.
- 15. A method of producing methionine comprising culturing a microorganism which has decreased RXA00655 activity under conditions such that methionine is produced.
- 16. A method of producing methionine comprising culturing a microorganism which overexpresses RXN02910 and a microorganism which has suppressed expression of RXA00655 under conditions such that methionine is produced.
- 17. A method of producing methionine comprising culturing a microorganism which overexpresses RXN02910 and which has suppressed expression of RXA00655 under conditions such that methionine is produced.
- 18. A method of producing methionine comprising culturing a microorganism containing a functionally disrupted RXA00655 gene under conditions such that methionine is produced.
- 19. A method of producing methionine comprising culturing a microorganism which has been transformed with a plasmid comprising the nucleotide sequence of SEQ ID NO: 13 under conditions such that methionine is produced.
- 20. A method of producing methionine comprising culturing a microorganism which has been transformed with a plasmid comprising the nucleotide sequence of SEQ ID NO: 3 under conditions such that methionine is produced.
- 21. The method of any of claims 1, 6, 12, 13, 14, 15, 16, 17, 18, 19 or 20, wherein said microorganism is Corynebacterium glutamicum.
- 22. The method of any one of claims 1, 6, 12, 13, 14, 15, 16, 17, 18, 19 or 20, further comprising recovering said methionine.
- 23. A method of modulating production of methionine by a microorganism comprising culturing a microorganism which overexpresses RXN02910 under conditions such that production of is methionine modulated.
- 24. A method of modulating production of methionine comprising culturing a microorganism which has suppressed expression of RXA00655 under conditions such that production of methionine is modulated.
- 25. A method of modulating production of methionine comprising culturing a microorganism which has increased RXN02910 activity under conditions such that production of methionine is modulated.
- 26. A method of modulating production of methionine comprising culturing a microorganism which has decreased RXA00655 activity under conditions such that production of methionine is modulated.
- 27. A method of modulating production of methionine comprising culturing a microorganism containing a functionally disrupted RXA00655 gene under conditions such that production of methionine is modulated.
- 28. A method of modulating production of methionine comprising culturing a microorganism which has been transformed with a plasmid comprising the nucleotide sequence of SEQ ID NO: 13 under conditions such that production of methionine is modulated.
- 29. A method of modulating production of methionine comprising culturing a microorganism which has been transformed with a plasmid comprising the nucleotide sequence of SEQ ID NO: 3 under conditions such that production of methionine is modulated.
- 30. The method of any one of claims 23-29, wherein production of methionine has been increased.
- 31. A method for producing a microorganism deficient in RXA00655 comprising transforming said microorganism with a plasmid comprising SEQ ID NO: 3.
- 32. A method for producing a microorganism which overexpresses RXA02910 comprising transforming said microorganism with a plasmid comprising SEQ ID NO: 13.
- 33. An isolated a nucleic acid molecule comprising the nucleotide sequence set forth in SEQ ID NO: 1, SEQ ID NO: 5, SEQ ID NO: 16, SEQ ID NO: 18, SEQ ID NO: 20, or SEQ ID NO: 22.
- 34. An isolated nucleic acid molecule which encodes a polypeptide comprising the amino acid sequence set forth in SEQ ID NO: 2, SEQ ID NO: 6, SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 21, or SEQ ID NO: 23.
- 35. An isolated nucleic acid molecule which encodes a naturally occurring allelic variant of a polypeptide comprising the amino acid sequence set forth in SEQ ID NO: 2, SEQ ID NO: 6, SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 21, or SEQ ID NO: 23.
- 36. An isolated nucleic acid molecule selected from the group consisting of:
a) a nucleic acid molecule comprising a nucleotide sequence which is at least 60% identical to the nucleotide sequence of SEQ ID NO: 1, SEQ ID NO: 5, SEQ ID NO: 16, SEQ ID NO: 18, SEQ ID NO: 20, or SEQ ID NO: 22, or a complement thereof; b) a nucleic acid molecule comprising a fragment of at least 30 nucleotides of a nucleic acid comprising the nucleotide sequence of SEQ ID NO: 1, SEQ ID NO: 5, SEQ ID NO: 16, SEQ ID NO: 18, SEQ ID NO: 20, or SEQ ID NO: 22, or a complement thereof; c) a nucleic acid molecule which encodes a polypeptide comprising an amino acid sequence at least about 60% identical to the amino acid sequence of SEQ ID NO: 2, SEQ ID NO: 6, SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 21, or SEQ ID NO: 23; and d) a nucleic acid molecule which encodes a fragment of a polypeptide comprising the amino acid sequence of SEQ ID NO: 2, SEQ ID NO: 6, SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 21, or SEQ ID NO: 23, wherein the fragment comprises at least 10 contiguous amino acid residues of the amino acid sequence of SEQ ID NO: 2, SEQ ID NO: 6, SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 21, or SEQ ID NO: 23.
- 37. An isolated nucleic acid molecule which hybridizes to a complement of the nucleic acid molecule of any one of claims 33, 34, 35, or 36, under stringent conditions.
- 38. An isolated nucleic acid molecule comprising a nucleotide sequence which is complementary to the nucleotide sequence of the nucleic acid molecule of any one of claims 33, 34, 35, or 36.
- 39. An isolated nucleic acid molecule comprising the nucleic acid molecule of any one of claims 33, 34, 35, or 36, and a nucleotide sequence encoding a heterologous polypeptide.
- 40. A vector comprising the nucleic acid molecule of any one of claims 33, 34, 35, or 36.
- 41. The vector of claim 40, which is an expression vector.
- 42. A host cell transfected with the expression vector of claim 41.
- 43. A method of producing a polypeptide comprising culturing the host cell of claim 42 in an appropriate culture medium to, thereby, produce the polypeptide.
- 44. An isolated polypeptide selected from the group consisting of:
a) a fragment of a polypeptide comprising the amino acid sequence of SEQ ID NO: 2, SEQ ID NO: 6, SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 21, or SEQ ID NO: 23, wherein the fragment comprises at least 10 contiguous amino acids of SEQ ID NO: 2, SEQ ID NO: 6, SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 21, or SEQ ID NO: 23; b) a naturally occurring allelic variant of a polypeptide comprising the amino acid sequence of SEQ ID NO: 2, SEQ ID NO: 6, SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 21, or SEQ ID NO: 23, wherein the polypeptide is encoded by a nucleic acid molecule which hybridizes to a complement of a nucleic acid molecule consisting of SEQ ID NO: 1, SEQ ID NO: 5, SEQ ID NO: 16, SEQ ID NO: 18, SEQ ID NO: 20, or SEQ ID NO: 22 under stringent conditions; c) a polypeptide which is encoded by a nucleic acid molecule comprising a nucleotide sequence which is at least 60% identical to a nucleic acid comprising the nucleotide sequence of SEQ ID NO: 1, SEQ ID NO: 5, SEQ ID NO: 16, SEQ ID NO: 18, SEQ ID NO: 20, or SEQ ID NO: 22; and d) a polypeptide comprising an amino acid sequence which is at least 60% identical to the amino acid sequence of SEQ ID NO: 2, SEQ ID NO: 6, SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 21, or SEQ ID NO: 23.
- 45. The isolated polypeptide of claim 44 comprising the amino acid sequence of SEQ ID NO: 2, SEQ ID NO: 6, SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 21, or SEQ ID NO: 23.
- 46. The polypeptide of claim 44, further comprising heterologous amino acid sequences.
- 47. A method for producing methionine, comprising culturing a microorganism containing a vector of claim 40 such that methionine is produced.
- 48. The method of claim 47, wherein said method further comprises the step of recovering methionine from said culture.
- 49. The method of claim 47, wherein said microorganism belongs to the genus Corynebacterium or Brevibacterium.
- 50. The method of claim 47, wherein said microorganism is selected from the group consisting of: Corynebacterium glutamicum, Corynebacterium efficiens, Corynebacterium herculis, Corynebacterium, lilium, Corynebacterium acetoacidophilum, Corynebacterium acetoglutamicum, Corynebacterium acetophilum, Corynebacterium ammoniagenes, Corynebacterium fujiokense, Corynebacterium nitrilophilus, Brevibacterium ammoniagenes, Brevibacterium butanicum, Brevibacterium divaricatum, Brevibacterium flavum, Brevibacterium healii, Brevibacterium ketoglutamicum, Brevibacterium ketosoreductum, Brevibacterium lactofermentum, Brevibacterium linens, Brevibacterium paraffinolyticum, and those strains set forth in Table 1.
- 51. The method of claim 47, wherein expression of the nucleic acid molecule from said vector results in modulation of production of said methionine.
- 52. A method of increasing production of a sulfur-containing compound by a microorganism comprising culturing a microorganism which overexpresses a positive regulator of methionine biosynthesis under conditions such that production of said sulfur-containing compound is increased.
- 53. The method of claim 52, wherein said sulfur-containing compound is selected from the group consisting of methionine, cysteine, S-adenosylmethionine and homocycsteine.
- 54. The method of claim 52, wherein said positive regulator of methionine biosynthesis is RXN02910.
- 55. The method of claim 52, wherein RXN02910 comprises a nucleic acid molecule comprising the nucleotide sequence set forth as SEQ ID NO: 5 or SEQ ID NO: 16.
- 56. The method of claim 52, wherein said microorganism has been transformed with a plasmid comprising the nucleotide sequence of SEQ ID NO: 13.
- 57. The method of claim 52, wherein said positive regulator of methionine biosynthesis is a transcriptional regulatory protein.
- 58. A method of increasing production of a sulfur-containing compound by a microorganism comprising culturing a microorganism which underexpresses a negative regulator of methionine biosynthesis under conditions such that production of said sulfur-containing compound is increased.
- 59. The method of claim 58, wherein said sulfur-containing compound is selected from the group consisting of methionine, cysteine, S-adenosylmethionine and homocycsteine.
- 60. The method of claim 58, wherein said negative regulator of methionine biosynthesis is RXN00655.
- 61. The method of claim 58, wherein RXN00655 comprises a nucleic acid molecule comprising the nucleotide sequence set forth as SEQ ID NO: 1.
- 62. The method of claim 58, wherein said microorganism has been transformed with a plasmid comprising the nucleotide sequence of SEQ ID NO: 3.
- 63. The method of claim 58, wherein said negative regulator of methionine biosynthesis is a transcriptional regulatory protein.
Priority Claims (12)
Number |
Date |
Country |
Kind |
19930476.9 |
Jul 1999 |
DE |
|
19931419.5 |
Jul 1999 |
DE |
|
19931420.9 |
Jul 1999 |
DE |
|
19932122.1 |
Jul 1999 |
DE |
|
19932128.0 |
Jul 1999 |
DE |
|
19932134.5 |
Jul 1999 |
DE |
|
19932206.6 |
Jul 1999 |
DE |
|
19932207.4 |
Jul 1999 |
DE |
|
19933003.4 |
Jul 1999 |
DE |
|
19941390.8 |
Aug 1999 |
DE |
|
19942088.2 |
Sep 1999 |
DE |
|
19942124.2 |
Sep 1999 |
DE |
|
RELATED APPLICATIONS
[0001] This application claims priority to U.S. Provisional Application Serial No. 60/422,618, filed on Oct. 30, 2002. This application also claims priority to U.S. application Ser. No. 09/602,874, filed on Jun. 23, 2000. This application also claims priority to U.S. Provisional Patent Application No. 60/141031, filed Jun. 25, 1999, U.S. Provisional Patent Application No. 60/142690, filed Jul. 1, 1999, and also to U.S. Provisional Patent Application No. 60/151251, filed Aug. 27, 1999. This application also claims priority to German Patent Application No. 19930476.9, filed Jul. 1, 1999, German Patent Application No. 19931419.5, filed Jul. 8, 1999, German Patent Application No. 19931420.9, filed Jul. 8, 1999, German Patent Application No. 19932122.1, filed Jul. 9, 1999, German Patent Application No. 19932128.0, filed Jul. 9, 1999, German Patent Application No. 19932134.5, filed Jul. 9, 1999, German Patent Application No. 19932206.6, filed Jul. 9, 1999, German Patent Application No. 19932207.4, filed Jul. 9, 1999, German Patent Application No. 19933003.4, filed Jul. 14, 1999, German Patent Application No. 19941390.8, filed Aug. 31, 1999, German Patent Application No. 19942088.2, filed Sep. 3, 1999, and German Patent Application No. 19942124.2, filed Sep. 3, 1999. The entire contents of all of the aforementioned applications are hereby expressly incorporated herein by this reference.
Provisional Applications (4)
|
Number |
Date |
Country |
|
60141031 |
Jun 1999 |
US |
|
60142690 |
Jul 1999 |
US |
|
60151251 |
Aug 1999 |
US |
|
60422618 |
Oct 2002 |
US |
Continuation in Parts (1)
|
Number |
Date |
Country |
Parent |
09602874 |
Jun 2000 |
US |
Child |
10307138 |
Nov 2002 |
US |