PUBLICATION

From discovery to potential application: engineering a novel M23 peptidase to combat Listeria monocytogenes

Authors
Kaus-Drobek, M., Nowacka, M., Gewartowska, M., Korzeniowska Nee Wiweger, M., Jensen, M.R., Møretrø, T., Heir, E., Nowak, E., Sabała, I.
ID
ZDB-PUB-250506-10
Date
2025
Source
Scientific Reports   15: 1562815628 (Journal)
Registered Authors
Keywords
Listeria monocytogenes, Antimicrobial agents, Biofilm eradication, M23 peptidases, Peptidoglycan hydrolases
MeSH Terms
  • Protein Engineering
  • Zebrafish
  • Peptide Hydrolases*/genetics
  • Peptide Hydrolases*/pharmacology
  • Animals
  • Microbial Sensitivity Tests
  • Anti-Bacterial Agents*/pharmacology
  • Bacterial Proteins*/genetics
  • Bacterial Proteins*/metabolism
  • Bacterial Proteins*/pharmacology
  • N-Acetylmuramoyl-L-alanine Amidase/genetics
  • N-Acetylmuramoyl-L-alanine Amidase/metabolism
  • N-Acetylmuramoyl-L-alanine Amidase/pharmacology
  • Listeria monocytogenes*/drug effects
  • Listeriosis/drug therapy
  • Listeriosis/microbiology
  • Biofilms/drug effects
  • Humans
PubMed
40325189 Full text @ Sci. Rep.
Abstract
Peptidoglycan hydrolases are promising alternatives for combating pathogens due to their specificity and potent bacteriolytic activity. In this study, a novel M23 peptidase from Streptococcus thermophilus NCTC10353, designated StM23, was discovered and characterized. It exhibited antibacterial activity against Listeria monocytogenes and other Gram-positive bacteria with meso-DAP-type peptidoglycan, including Bacillus subtilis and Bacillus cereus. To enhance StM23's efficacy and specificity, a chimeric enzyme, StM23_CWT, was engineered by fusing its catalytic domain with a cell wall-targeting domain (CWT) from SpM23B, a peptidoglycan hydrolase found in Staphylococcus pettenkoferi. The engineered chimera demonstrated expanded specificity, showing activity against Staphylococcus aureus and Enterococcus faecium. Its ability to disrupt L. monocytogenes cells was visualized by electron microscopy. The enzyme effectively disrupted biofilm structures and decontaminated surfaces like glass, stainless steel, and silicone, showcasing its industrial potential. Safety evaluations using zebrafish, moth larvae, and human cell models confirmed its non-toxic profile, supporting its broad applicability. Based on these findings, StM23_CWT is a novel and potent antimicrobial agent with significant potential to reduce the risk of listeriosis and control persistent pathogens.
Genes / Markers
Figures
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Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping