Isolation and characterisation of a novel antibacterial peptide from a native swine intestinal tract-derived bacterium

Int J Antimicrob Agents. 2017 Apr;49(4):427-436. doi: 10.1016/j.ijantimicag.2016.12.012. Epub 2017 Feb 27.

Abstract

Antimicrobial peptides (AMPs) are highly associated with antipathogenic activity, without generating drug resistance in targeted bacteria. In this study, the existence of AMPs in the Tibetan swine, a China-native, cold-resistant and seldom-sick breed of pig, was investigated. A peptide secreted by a Tibetan swine intestinal tract-derived Bacillus strain was isolated using reversed-phase chromatography (RPC), ultrafiltration and reversed-phase high-performance liquid chromatography (RP-HPLC). The peptide was identified by mass spectrometry and was characterised for activity against Escherichia coli and Staphylococcus aureus. The 16-amino acid peptide (ASVVNKLTGGVAGLLK), named TP, had a molecular mass of 1568.919 Da and exhibited inhibitory activity against Gram-positive and Gram-negative bacteria [minimum inhibitory concentrations (MICs) of 2.5-5 µM and 10-20 µM for E. coli and S. aureus, respectively] as well as human MKN-45 and NB4 tumour cell lines [50% inhibitory concentration (IC50) = 4.686 µM and 11.479 µM, respectively]. TP also exhibited weak haemolytic activity. Furthermore, TP enhanced cell membrane permeability and K+ outflow, bound with E. coli genomic DNA in vitro and inhibited E. coli growth. Thus, TP represents a strong candidate as an antibacterial peptide.

Keywords: Antibacterial peptide; Antitumour; MIC; Tibet swine.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Anti-Bacterial Agents / chemistry
  • Anti-Bacterial Agents / isolation & purification
  • Anti-Bacterial Agents / pharmacology*
  • Bacillus / chemistry*
  • Bacillus / isolation & purification
  • Biological Products / chemistry
  • Biological Products / isolation & purification
  • Biological Products / pharmacology*
  • China
  • Chromatography, High Pressure Liquid
  • Chromatography, Reverse-Phase
  • Escherichia coli / drug effects*
  • Gastrointestinal Tract / microbiology*
  • Mass Spectrometry
  • Microbial Sensitivity Tests
  • Molecular Weight
  • Peptides / chemistry
  • Peptides / isolation & purification
  • Peptides / pharmacology*
  • Staphylococcus aureus / drug effects*
  • Swine
  • Ultracentrifugation

Substances

  • Anti-Bacterial Agents
  • Biological Products
  • Peptides