Anti-biofilm and sporicidal activity of peptides based on wheat puroindoline and barley hordoindoline proteins

J Pept Sci. 2016 Jul;22(7):492-500. doi: 10.1002/psc.2895. Epub 2016 May 30.

Abstract

The broad-spectrum activity of antimicrobial peptides (AMPs) and low probability of development of host resistance make them excellent candidates as novel bio-control agents. A number of AMPs are found to be cationic, and a small proportion of these are tryptophan-rich. The puroindolines (PIN) are small, basic proteins found in wheat grains with proposed roles in biotic defence of seeds and seedlings. Synthetic peptides based on their unique tryptophan-rich domain (TRD) display antimicrobial properties. Bacterial endospores and biofilms are highly resistant cells, with significant implications in both medical and food industries. In this study, the cationic PIN TRD-based peptides PuroA (FPVTWRWWKWWKG-NH2 ) and Pina-M (FSVTWRWWKWWKG-NH2 ) and the related barley hordoindoline (HIN) based Hina (FPVTWRWWTWWKG-NH2 ) were tested for effects on planktonic cells and biofilms of the common human pathogens including Pseudomonas aeruginosa, Listeria monocytogenes and the non-pathogenic Listeria innocua. All peptides showed significant bactericidal activity. Further, PuroA and Pina-M at 2 × MIC prevented initial biomass attachment by 85-90% and inhibited >90% of 6-h preformed biofilms of all three organisms. However Hina, with a substitution of Lys-9 with uncharged Thr, particularly inhibited Listeria biofilms. The PIN based peptides were also tested against vegetative cells and endospores of Bacillus subtilis. The results provided evidence that these tryptophan-rich peptides could kill B. subtilis even in sporulated state, reducing the number of viable spores by 4 log units. The treated spores appeared withered under scanning electron microscopy. The results establish the potential of these tryptophan-rich peptides in controlling persistent pathogens of relevance to food industries and human health. Copyright © 2016 European Peptide Society and John Wiley & Sons, Ltd.

Keywords: Trp-rich peptides; antimicrobial peptides; biofilms; hordoindolines; puroindolines; tryptophan.

MeSH terms

  • Amino Acid Sequence
  • Anti-Bacterial Agents / chemistry
  • Anti-Bacterial Agents / isolation & purification
  • Anti-Bacterial Agents / pharmacology*
  • Antimicrobial Cationic Peptides / chemistry
  • Antimicrobial Cationic Peptides / isolation & purification
  • Antimicrobial Cationic Peptides / pharmacology*
  • Bacillus subtilis / drug effects
  • Bacillus subtilis / growth & development
  • Bacillus subtilis / ultrastructure
  • Biofilms / drug effects*
  • Biofilms / growth & development
  • Hordeum / chemistry
  • Hordeum / immunology
  • Listeria / drug effects
  • Listeria / growth & development
  • Listeria monocytogenes / drug effects
  • Listeria monocytogenes / growth & development
  • Microbial Sensitivity Tests
  • Plankton / drug effects
  • Plankton / growth & development
  • Plant Proteins / chemistry
  • Plant Proteins / isolation & purification
  • Plant Proteins / pharmacology*
  • Pseudomonas aeruginosa / drug effects
  • Pseudomonas aeruginosa / growth & development
  • Spores, Bacterial / drug effects*
  • Spores, Bacterial / growth & development
  • Spores, Bacterial / ultrastructure
  • Triticum / chemistry
  • Triticum / immunology
  • Tryptophan / chemistry

Substances

  • Anti-Bacterial Agents
  • Antimicrobial Cationic Peptides
  • Plant Proteins
  • hordoindoline B protein, Hordeum vulgare
  • puroindoline protein, Triticum aestivum
  • Tryptophan