Glutamate 2,3-aminomutases and methods of use thereof
There are provided nucleic acids, including isolated DNA molecules, which encode glutamate 2,3-aminomutase enzymes, polypeptides produced from such nucleic acids and methods of making the nucleic acids and polypeptides. There are further provided methods of producing β-glutamate from glutamate using glutamate 2,3-aminomutase.
1. An isolated DNA molecule encoding a polypeptide having glutamate 2,3-aminomutase activity, wherein the DNA molecule comprises a sequence selected from the group consisting of
a) a DNA sequence encoding a polypeptide having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 2, wherein the polypeptide comprises the following amino acid sequences: CXXXCRXCXR (SEQ ID NO: 19), S/(T)GGE/(D) (SEQ ID NO: 20), GXXXPXXXXXXXXXXXK (SEQ ID NO: 21), PXYXXXXKXXXG (SEQ ID NO: 22), and PXXXXNXXXXXXK (SEQ ID NO: 23);
b) a DNA sequence comprising SEQ ID NO: 1; and
c) a DNA sequence which is degenerate to the sequence of (b) due to the genetic code.
2. An expression vector comprising the DNA molecule of claim 1 .
3. The expression vector of claim 2 wherein the DNA molecule is operably linked to one or more control sequences which direct the production of the polypeptide in a host cell.
4. A recombinant host cell comprising the DNA molecule of claim 1 .
5. A recombinant host cell comprising the expression vector of claim 2 .
6. The recombinant host cell of claim 5 wherein the recombinant host cell is a prokaryotic cell.
7. The recombinant host cell of claim 6 wherein the cell is a bacterial cell.
8. The recombinant host cell of claim 6 wherein the cell is an E. coli cell.
9. A method of producing a polypeptide having glutamate 2,3-aminomutase activity comprising culturing the recombinant host cell of claim 6 .
10. The method of claim 9 further comprising isolating the polypeptide produced from the recombinant host cell.
11. The isolated DNA molecule of claim 1 wherein the DNA sequence encoding the polypeptide has at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 2.
12. An isolated DNA molecule encoding a polypeptide having glutamate 2,3-aminomutase activity, wherein the DNA molecule comprises a sequence encoding a polypeptide having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 2.
13. A recombinant host cell comprising the DNA molecule of claim 12 .
14. An expression vector comprising the DNA molecule of claim 12 .
15. The expression vector of claim 14 wherein the DNA molecule is operably linked to one or more control sequences which direct the production of the polypeptide in a host cell.
16. A recombinant host cell comprising the expression vector of claim 15 .
17. The recombinant host cell of claim 16 wherein the recombinant host cell is a prokaryotic cell.
18. The recombinant host cell of claim 17 wherein the cell is a bacterial cell.
19. The recombinant host cell of claim 18 wherein the cell is an E. coli cell.
20. A method of producing a polypeptide having glutamate 2,3-aminomutase activity comprising culturing the recombinant host cell of claim 16 .
21. The method of claim 20 further comprising isolating the polypeptide produced from the recombinant host cell.