Alleles of the rel gene from coryneform bacteria
View Patent ↗An isolated mutant of a coryneform bacterium comprising a gene coding for a polypeptide having GTP-pyrophosphate kinase activity, wherein said polypeptide comprises an amino acid sequence in which one of the proteinogenic amino acids other than L-proline is present in position 38 or a corresponding or comparable position. In addition, an isolated polynucleotide encoding a polypeptide having GTP-pyrophosphate kinase enzyme activity, a vector comprising the isolated polynucleotide, a recombinant microorganism comprising the vector, and a process for preparing the recombinant coryneform bacterium is described. A method for over-expressing a GTP-pyrophosphate kinase, a method of preparing an L-amino acid, an L-lysine comprising and L-tryptophan comprising feed is also described.
1. An isolated polynucleotide encoding an amino acid sequence comprising a polypeptide having GTP-pyrophosphate kinase enzyme activity, wherein the polypeptide is as set forth by SEQ ID NO: 2 in which L-proline in position 38 has been replaced by L-leucine.
2. The isolated polynucleotide as claimed in claim 1 , wherein the encoded amino acid sequence is 760 amino acids in length.
3. The isolated polynucleotide as claimed in claim 1 , which comprises nucleotides 751-3033 of SEQ ID NO: 7.
4. The isolated polynucleotide as claimed in claim 1 , which hybridizes with the nucleotide sequence complementary to the full-length of SEQ ID NO: 5 under conditions of hybridization in 5×SSC at approximately 50° C.-68° C. followed by washing in 0.5×SSC at 66° C.-68° C.
5. A process for preparing a recombinant microorganism, comprising
a) transferring the isolated polynucleotide as claimed in claim 1 to a microorganism,
b) replicating said isolated polynucleotide in said microorganism, and
c) propagating the microorganism obtained by a) and b).
6. A recombinant microorganism comprising the isolated polynucleotide as claimed in claim 1 .
7. The recombinant microorganism as claimed in claim 6 , which is a coryneform bacterium or a bacterium of the genus Escherichia.
8. The recombinant microorganism as claimed in claim 7 , wherein the coryneform bacterium is the genus Corynebacterium.
9. The recombinant microorganism as claimed in claim 8 , wherein the bacterium of the genus Corynebacterium is the species Corynebacterium glutamicum.
10. A vector comprising the isolated polynucleotide as claimed in claim 1 .
11. A recombinant microorganism comprising the vector as claimed in claim 10 .
12. The recombinant microorganism as claimed in claim 11 , which is a coryneform bacterium or a bacterium of the genus Escherichia.
13. The recombinant microorganism as claimed in claim 12 , wherein the coryneform bacterium is the genus Corynebacterium.
14. The recombinant microorganism as claimed in claim 13 , wherein the bacterium of the genus Corynebacterium is the species Corynebacterium glutamicum.
15. A process for overexpressing a GTP-pyrophosphate kinase in a microorganism comprising increasing the copy number of the polynucleotide as claimed in claim 1 in a microorganism by at least one (1) copy.
16. A process for overexpressing a GTP-pyrophosphate kinase in a microorganism, comprising functionally linking a promotor or an expression cassette to the polynucleotide of claim 1 and expressing the encoded polypeptide in a microorganism.