Isolated nucleic acid molecules, designated SMP nucleic acid molecules, which encode novel SMP proteins from Corynebacterium glutamicum are described. The invention also provides antisense nucleic acid molecules, recombinant expression vectors containing SMP nucleic acid molecules, and host cells into which the expression vectors have been introduced. The invention still further provides isolated SMP proteins, mutated SMP proteins, fusion proteins, antigenic peptides and methods for the improvement of production of a desired compound from C. glutamicum based on genetic engineering of SMP genes in this organism.
1. An isolated nucleic acid molecule
comprising the nucleotide sequence of SEQ ID NO:179, or the complement thereof.
2. An isolated nucleic acid molecule comprising a nucleotide sequence which is at least 95% identical to the entire nucleotide sequence of SEQ ID NO:179, wherein the nucleic acid molecule encodes a polypeptide having a phosphoenolpyruvate carboxykinase activity, or the complement thereof.
3. An isolated nucleic acid molecule comprising a nucleotide sequence which is at least 98% identical to the entire nucleotide sequence of SEQ ID NO:179, wherein the nucleic acid molecule encodes a polypeptide having a phosphoenolpyruvate carboxykinase activity, or the complement thereof.
4. An isolated nucleic acid molecule which encodes a polypeptide comprising the amino acid sequence of SEQ ID NO:180, or the complement thereof.
5. An isolated nucleic acid molecule comprising a fragment of at least 25 contiguous nucleotides of the nucleotide sequence of SEQ ID NO:179, wherein the fragment encodes a polypeptide having a phosphoenolpyruvate carboxykinase activity, or the complement thereof.
6. An isolated nucleic acid molecule consisting of a fragment of at least 25 contiguous nucleotides of the nucleotide sequence of SEQ ID NO:179, or the complement thereof.
7. An isolated nucleic acid molecule which encodes a polypeptide comprising an amino acid sequence which is at least 95% identical to the entire amino acid sequence of SEQ ID NO:180, wherein the polypeptide has a phosphoenolpyruvate carboxykinase activity, or the complement thereof.
8. An isolated nucleic acid molecule which encodes a polypeptide comprising an amino acid sequence which is at least 98% identical to the entire amino acid sequence of SEQ ID NO:180, wherein the polypeptide has a phosphoenolpyruvate carboxykinase activity, or the complement thereof.
9. An isolated nucleic acid molecule comprising the nucleic acid molecule of any one of claims 1 or 2 - 8 and a nucleotide sequence encoding a heterologous polypeptide.
10. A vector comprising the nucleic acid molecule of any one of claims 1 or 2 - 8 .
11. The vector of claim 10 , which is an expression vector.
12. A host cell transfected with the expression vector of claim 11 , wherein said host cell is a microorganism.
13. The host cell of claim 12 , wherein said cell is a bacterial cell.
14. The host cell of claim 13 , wherein said cell belongs to the genus Corynebacterium or Brevibactertum.
15. A method of producing a polypeptide comprising culturing the host cell of claim 12 in an appropriate culture medium to, thereby, produce the polypeptide.
16. A method for producing an amino acid, comprising culturing the cell of claim 12 such that the amino acid is produced.
17. The method of claim 16 , wherein said method further comprises the step of recovering the amino acid from said culture.
18. The method of claim 16 , wherein said cell belongs to the genus Corynebactertium or Brevibacterium.
19. The method of claim 16 , wherein said cell is selected from the group consisting of Corynebacterium glutamicum, Corynebacterium herculis, Corynebacterium lilium, Corynebactertium 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 3.
20. The metbod of claim 16 , wherein expression of the nucleic acid molecule from said vector results in modulation of production of said amino acid.
21. The method of claim 16 , wherein said anino acid is selected from the group consisting of lysine, glutamate, glutamine, alanine, aspartate, glycine, serine, threonine, methionine, cysteine, valine, leucine, isoleucine, arginine, proline, histidine, tyrosine, phenylalanine, and tryptophan.
22. A method for diagnosing the presence or activity of Corynebacterium diphtheriae in a subject, comprising detecting the presence of at least one of the nucleic acid molecules of any one of claims 1 or 2 - 8 , thereby diagnosing the presence or activity of Corynebacterium diphtheriae in the subject.