Mutated penicillin G acylase genes
OwnerJan M. Van Der Laan, Adriana M. Riemens +1inventors
InventorsJan M. Van Der Laan, Adriana M. Riemens, Wilhelmus J. Quax
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Abstract
New mutant .beta.-lactam Penicillin G acylases are provided, exhibiting altered substrate specificities. These Penicillin G acylases are obtained by expression of a gene encoding said Penicillin G acylase and having an amino acid sequence which differs at least in one amino acid from the wild-type Penicillin G acylase.
Claims13
An isolated mutant prokaryotic Penicillin G acylase or its preenzmye or preproenzyme having:
- an amino acid substitution at one or more of the positions corresponding to A139 to A142 and A148 to A152 as set forth in SEQ ID NO:
- 27, B20 to B27, B31, B49 to B52, B56, B57, B65, B67 to B72, B154 to B157, B173 to B179, B239 to B241, B250 to B263, B379 to B387, B390, B455, B474 to B480 as set forth in SEQ ID NO:32 in Alcaligenes faecalis Penicillin G acylase or its pre- or preproenzyme, wherein the substitution at position A141 results in a substituent amino acid selected from the group consisting of Ala, Arg, Asn, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, and Val;
- and
- an altered substrate specificity or altered specific activity relative to the corresponding wild-type unsubstituted Penicillin G acylase.
An isolated mutant prokaryotic Penicillin G acylase or its preenzyme or preproenzyme having an amino acid substitution at a position corresponding to A141 (Gly) as set forth in SEQ ID NO:
- 27 wherein A141 (Gly) is changed to Cys, Thr, Met, Ala, Val or Leu.
An isolated mutant prokaryotic Penicillin G acylase or its preenzyme or preproenzyme having an amino acid substitution at a position corresponding to B24 (Phe) as set forth in SEQ ID NO:
- 32 wherein B24 (Phe) is changed to Ala or Leu.
Dependencies are inferred from the claim text.