Modified beta-lactamases and methods and uses related thereto
The present invention relates to pharmaceuticals and modified beta-lactamases. Specifically, the invention relates to novel recombinant beta-lactamases and pharmaceutical compositions comprising the beta-lactamases. Also, the present invention relates to methods for modifying a beta-lactamase, producing the beta-lactamase and treating or preventing beta-lactam antibiotic induced adverse effects. Furthermore, the present invention relates to the beta-lactamase for use as a medicament and to the use of the beta-lactamase in the manufacture of a medicament for treating or preventing beta-lactam antibiotics induced adverse effects. Still further, the invention relates to a polynucleotide and a host cell comprising the polynucleotide.
1. A beta-lactamase, comprising an amino acid sequence having at least 90% sequence identity with SEQ ID NO: 1 and a hydrophilic amino acid residue other than aspartic acid (D) at a position corresponding to position 276 according to Ambler classification, wherein:
the hydrophilic amino acid residue is asparagine (N) and
the beta-lactamase hydrolyzes ceftriaxone substantially more efficiently than a beta-lactamase of SEQ ID NO: 1 that has an aspartic acid (D) at a position corresponding to position 276 according to Ambler classification.
2. The beta-lactamase according to claim 1 , wherein the hydrophilic amino acid residue is located in an alpha helix.
3. The beta-lactamase according to claim 1 , wherein the beta-lactamase further comprises at least one amino acid selected from a leucine (L) at a position corresponding to position 220 and an arginine (R) at a position corresponding to position 244 according to Ambler classification.
4. The beta-lactamase according to claim 1 , wherein the beta-lactamase hydrolyzes a penicillin and a cephalosporin.
5. The beta-lactamase according to claim 4 , wherein the cephalosporin is selected from cefoperazone and ceftriaxone.
6. The beta-lactamase according to claim 1 , wherein the amino acid sequence has at least 93% sequence identity with SEQ ID NO: 1.
7. The beta-lactamase according to claim 1 , wherein the amino acid sequence has at least 95% sequence identity with SEQ ID NO: 1.
8. A pharmaceutical composition for oral administration comprising an effective amount of a beta-lactamase comprising an amino acid sequence having at least 90% sequence identity with SEQ ID NO: 1 and a hydrophilic amino acid residue other than aspartic acid (D) at a position corresponding to position 276 according to Ambler classification, wherein:
the hydrophilic amino acid residue is asparagine (N) and
the beta-lactamase hydrolyzes ceftriaxone substantially more efficiently than a beta-lactamase of SEQ ID NO: 1 that has an aspartic acid (D) at a position corresponding to position 276 according to Ambler classification.
9. The pharmaceutical composition according to claim 8 , wherein the beta-lactamase comprises an amino acid sequence of at least 95% sequence identity with SEQ ID NO: 1.
10. A pharmaceutical composition comprising the beta-lactamase according to claim 1 .
11. The pharmaceutical composition according to claim 8 , wherein the beta-lactamase comprises an amino acid sequence of at least 93% sequence identity with SEQ ID NO: 1.
12. The pharmaceutical composition according to claim 8 , wherein the beta-lactamase hydrolyzes a penicillin and a cephalosporin.
13. The pharmaceutical composition according to claim 12 , wherein the cephalosporin is selected from cefoperazone and ceftriaxone.
14. A beta-lactamase comprising an amino acid sequence of SEQ ID NO: 1 with an asparagine (N) residue at position 276 according to Ambler classification.
15. The beta-lactamase according to claim 14 , wherein the beta-lactamase hydrolyzes a cephalosporin and a penicillin.
16. The beta-lactamase according to claim 15 , wherein the cephalosporin is selected from cefoperazone and ceftriaxone.
17. The beta-lactamase according to claim 14 , wherein the beta-lactamase hydrolyzes a cephalosporin that has been excreted into the gastrointestinal tract.