Enterocin A mutants identified by saturation mutagenesis enhance potency towards vancomycin‐resistant Enterococci. Issue 2 (3rd September 2015)

• Record Type: Journal Article
• Authors: McClintock, Maria K.; Kaznessis, Yiannis N.; Hackel, Benjamin J.
• Is Part Of: Biotechnology and bioengineering. Volume 113:Issue 2(2016)
• Page Start: 414
• Page End: 423
• Publication Date: 2015-09-03
• Subjects: antimicrobial peptide -- enterocin A -- lactic acid bacteria -- peptide engineering -- saturation mutagenesis; Biotechnology -- Periodicals; Bioengineering -- Periodicals; 660.6
• DOI: 10.1002/bit.25710
• Languages: English
• ISSNs: 0006-3592

Abstract

ABSTRACT: Vancomycin‐resistant Enterococci infections are a significant clinical problem. One proposed solution is to use probiotics, such as lactic acid bacteria, to produce antimicrobial peptides at the site of infection. Enterocin A, a class 2a bacteriocin, exhibits inhibitory activity against E. faecium and E. faecalis, which account for 86% of vancomycin‐resistant Enterococci infections. In this study, we aimed to engineer enterocin A mutants with enhanced potency within a lactic acid bacterial production system. Peptide mutants resulting from saturation mutagenesis at sites A24 and T27 were efficiently screened in a 96‐well plate assay for inhibition of pathogen growth. Several mutants exhibit increased potency relative to wild‐type enterocin A in both liquid‐ and solid‐medium growth assays. In particular, A24P and T27G exhibit enhanced inhibition of multiple strains of E. faecium and E. faecalis, including clinically isolated vancomycin‐resistant strains. A24P and T27G enhance killing of E. faecium 8 by 13 ± 3‐ and 18 ± 4‐fold, respectively. The engineered enterocin A/lactic acid bacteria systems offer significant potential to combat antibiotic‐resistant infections. Biotechnol. Bioeng. 2016;113: 414–423. © 2015 Wiley Periodicals, Inc. Abstract : Vancomycin‐resistant Enterococci are a significant clinical problem for which antimicrobial peptides offer a possible solution. Enterocin A mutants, produced in lactic acid bacteria, were identified through saturation mutagenesis at sites A24 and T27 that increase (up to 18 ± 4‐fold) killing of multiple strains of E. faecium and E. faecalis, including clinically isolated vancomycin‐resistant strains.