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- Title
A single point mutation converts a glutaryl-7-aminocephalosporanic acid acylase into an N-acyl-homoserine lactone acylase.
- Authors
Murugayah, Shereen A.; Evans, Gary B.; Tyndall, Joel D. A.; Gerth, Monica L.
- Abstract
Objective: To change the specificity of a glutaryl-7-aminocephalosporanic acid acylase (GCA) towards N-acyl homoserine lactones (AHLs; quorum sensing signalling molecules) by site-directed mutagenesis. Results: Seven residues were identified by analysis of existing crystal structures as potential determinants of substrate specificity. Site-saturation mutagenesis libraries were created for each of the seven selected positions. High-throughput activity screening of each library identified two variants—Arg255Ala, Arg255Gly—with new activities towards N-acyl homoserine lactone substrates. Structural modelling of the Arg255Gly mutation suggests that the smaller side-chain of glycine (as compared to arginine in the wild-type enzyme) avoids a key clash with the acyl group of the N-acyl homoserine lactone substrate. Conclusions: Mutation of a single amino acid residue successfully converted a GCA (with no detectable activity against AHLs) into an AHL acylase. This approach may be useful for further engineering of 'quorum quenching' enzymes.
- Subjects
ACYL-homoserine lactones; AMINO acid residues; HIGH throughput screening (Drug development); ACYL group; SITE-specific mutagenesis; QUORUM sensing; GLYCINE
- Publication
Biotechnology Letters, 2021, Vol 43, Issue 7, p1467
- ISSN
0141-5492
- Publication type
Article
- DOI
10.1007/s10529-021-03135-9