Genes encoding carbon metabolism and energy-producing proteins
View Patent ↗The invention relates to novel nucleic acid molecules, to the use thereof for constructing genetically improved microorganisms and to methods for preparing fine chemicals, in particular amino acids, with the aid of said genetically improved microorganisms.
1. An isolated nucleic acid molecule
encoding the amino acid sequence set forth in SEQ ID NO:32, wherein either (i) the amino acid residue at position 158 of SEQ ID NO:32 is any amino acid except proline, or (ii) the amino acid residue at position 361 is any amino acid except serine, or a complement thereof.
2. The isolated nucleic acid molecule of claim 1 wherein the amino acid residue at position 158 of SEQ ID NO:32 is serine.
3. An isolated nucleic acid molecule which hybridizes to the complement of the nucleotide sequence set forth in SEQ ID NO:31 at 6X sodium chloride/sodium citrate (SSC) at 45° followed by one or more washes in 0.2X SSC, 0.1% SDS at 50–65° C., wherein the nucleic acid molecule (a) encodes any amino acid except proline at the position corresponding to nucleotide residues 572–574 of SEQ ID NO:31 or (b) encodes any amino acid except serine at the position corresponding to nucleotide residues 1181–1183 of SEQ ID NO:31, and wherein the nucleic acid molecule encodes a polypeptide having a 6-phosphogluconate dehydrogenase activity, or a complement thereof.
4. An isolated nucleic acid molecule comprising the nucleotide sequence set forth in SEQ ID NO:31, wherein the nucleic acid molecule comprises one or more nucleic acid modifications at either (i) nucleotide residues 572–574 of SEQ ID NO:31 such that nucleotide residues 572–574 of SEQ ID NO:31 encode any amino acid except for proline or (ii) nucleotide residues 1181–1183 of SEQ ID NO:31 such that nucleotide residues 1181–1183 of SEQ ID NO:31 encode any amino acid except serine, or a complement thereof.
5. A vector comprising the nucleic acid molecule of any one of claims 1 , 3 or 4 .
6. The vector of claim 5 , which is an expression vector.
7. An isolated host cell, which is transfected with the vector of claim 6 .
8. The host cell of claim 7 , wherein expression of said nucleic acid molecule modulates the production of a fine chemical from said cell.
9. A method for preparing a fine chemical, comprising culturing the cell of claim 7 such that the fine chemical is produced.
10. The method of claim 9 , wherein the fine chemical is an amino acid.
11. The method of claim 10 , wherein said amino acid is lysine.
12. The host cell of claim 7 , wherein said cell is a microorganism.
13. The host cell of claim 12 , wherein said cell belongs to the genus Corynebacterium or Brevibacterium.
14. The host cell of claim 8 , wherein said fine chemical is selected from the group consisting of organic acids, proteinogenic and nonproteinogenic amino acids, purine and pyrimidine bases, nucleosides, nucleotides, lipids, saturated and unsaturated fatty acids, diols, carbohydrates, aromatic compounds, vitamins, cofactors and enzymes.
15. The method of claim 9 , wherein said cell belongs to the genus Corynebacterium or Brevibacterium.
16. The method of claim 9 , wherein expression of the nucleic acid molecule from said vector results in modulation of production of said fine chemical.
17. A method for producing a fine chemical, comprising culturing a cell whose genomic DNA has been altered by the inclusion of a nucleic acid molecule of any one of claims 1 , 3 or 4 .
18. The nucleic acid molecule of claim 4 , wherein nucleotide residues 572-574 of SEQ ID NO:31 encode serine.
19. The nucleic acid molecule of claim 4 , wherein nucleotide residues 1181-1183 of SEQ ID NO:31 encode phenylalanine.
20. The isolated nucleic acid molecule of claim 1 wherein the amino acid residue at position 361 of SEQ ID NO:32 is phenylalanine.
21. The nucleic acid molecule of claim 3 , wherein the nucleic acid molecule encodes phenylalanine at the position corresponding to nucleotide residues 1181-1183.
22. The nucleic acid molecule of claim 3 , wherein the nucleic acid molecule encodes serine at the position corresponding to nucleotide residues 572-574.