Nucleotide sequences which code for the ccpA2 gene
View Patent ↗The invention relates to polynucleotides corresponding to the ccpA2 gene and which encode a CcpA2 catabolite control protein, methods of producing L-amino acids, and methods of screening for polynucleotides which encode proteins having CcpA2 catabolite control activity.
1. A process for producing an L-amino acid comprising:
culturing a bacterial cell in a medium suitable for producing L-amino acid, and
recovering or isolating said L-amino acid,
wherein said bacterial cell has been modified to eliminate or reduce the expression of the CcpA2 polypeptide compared to the corresponding unmodified bacterium, and
wherein said CcpA2 polypeptide exhibits catabolite control activity and is encoded by a polynucleotide which is at least 95% identical to SEQ ID NO: 1.
2. The process of claim 1 , wherein said bacterial cell is a coryneform bacterium.
3. The process of claim 2 , wherein said bacterial cell is selected from the group consisting of Corynebacterium glutamicum, Corynebacterium acetoglutamicum, Corynebacterium acetoacidophilum, Corynebacterium melassecola, Corynebacterium thermoaminogenes, Brevibacterium flavum, Brevibacterium lactofermentum , and Brevibacterium divaricatum.
4. The process of claim 1 , wherein the CcpA2 polypeptide comprises SEQ ID NO: 2.
5. The process of claim 1 , wherein the L-amino acid is L-lysine.
6. The process of claim 1 , wherein the L-amino acid is not L-lysine.
7. The process of claim 1 , wherein the expression of the CcpA2 polypeptide has been eliminated.
8. The process of claim 1 , wherein the expression of the CcpA2 polypeptide has been attenuated.
9. The process of claim 1 , wherein the expression of the CcpA2 polypeptide has been attenuated by genetic modification of a signal structure involved in expression of the CcpA2 coding sequence.
10. The process of claim 1 , wherein the expression of the CcpA2 polypeptide has been eliminated or attenuated by modification of the CcpA2 coding sequence.
11. The process of claim 1 , wherein said bacterium contains one or more coding sequences from lysC, dapA, eno, zwf, dapD, dapE, gap, pyc, mqo, zal, or lysE, wherein said one or more coding sequences is over-expressed compared to a corresponding unmodified bacterium.
12. The process of claim 1 , wherein said bacterium has had one or more coding sequences from pck, pgi, poxB, or zwa2 eliminated or attenuated compared to a corresponding unmodified strain.
13. The process of claim 1 , wherein said L-amino acid is produced by a batch fermentation process.
14. The process of claim 1 , wherein said L-amino acid is produced by a fed batch fermentation process.
15. The process of claim 1 , wherein said L-amino acid is produced by a repeated fed fermentation process.
16. The process of claim 1 , wherein said L-amino acid is recovered from the culture medium.
17. The process of claim 1 , wherein said L-amino acid is recovered along with at least a portion of the biomass of the culture medium.
18. A process for producing an L-amino acid comprising:
culturing a bacterial cell in a medium suitable for producing an L-amino acid, and
recovering or isolating said L-amino acid;
wherein said bacterial cell has been modified to eliminate or reduce the expression of the CcpA2 polypeptide compared to the corresponding unmodified bacterium, and
wherein said CcpA2 polypeptide is encoded by a polynucleotide which may be obtained from Brevibacterium or Corynebacterium by PCR using the oligonucleotides of SEQ ID NOS: 3 and 4.
19. The process of claim 1 , wherein said CcpA2 polypeptide is encoded by a polynucleotide which comprises nucleotides 241 to 1278 of SEQ ID NO: 1.