Glycoside hydrolase (PelAh) immobilization prevents Pseudomonas aeruginosa biofilm formation on cellulose-based wound dressing

Carbohydr Polym. 2020 Oct 15:246:116625. doi: 10.1016/j.carbpol.2020.116625. Epub 2020 Jun 12.

Abstract

Bacterial cellulose (BC) is recognized as a wound dressing material well-suited for chronic wounds; however, it has no intrinsic antimicrobial activity. Further, the formation of biofilms can limit the effectiveness of the pre-saturation of BC with antimicrobial agents. Here, to hinder biofilm formation by P. aeruginosa, we immobilized the hydrolytic domain of PelA (a glycohydrolase involved in the synthesis of biofilm polysaccharide Pel) on the surface of BC. The immobilization of 32.35 ± 1.05 mg PelAh per g BC membrane resulted in an eight-fold higher P. aeruginosa cell detachment from BC membrane, indicating reduced biofilm matrix stability. Further, 1D and 2D infrared spectroscopy analysis indicated systematic reduction of polysaccharide biofilm elements, confirming the specificity of immobilized PelAh. Importantly, BC-PelAh was not cytotoxic towards L929 fibroblast cells. Thus, we conclude that PelAh can be used in BC wound dressings for safe and specific protection against biofilm formation by P. aeruginosa.

Keywords: Bacterial cellulose; Biofilm; PelA(h); Pseudomonas aeruginosa; Wound dressings.

MeSH terms

  • Acetobacteraceae / chemistry*
  • Acetobacteraceae / physiology
  • Animals
  • Anti-Bacterial Agents / metabolism
  • Anti-Bacterial Agents / pharmacology
  • Bacterial Proteins / biosynthesis
  • Bacterial Proteins / genetics
  • Bacterial Proteins / pharmacology
  • Bandages*
  • Biofilms / drug effects*
  • Biofilms / growth & development
  • Cell Line
  • Cellulose / biosynthesis
  • Cellulose / chemistry*
  • Cellulose / isolation & purification
  • Cloning, Molecular
  • Enzymes, Immobilized / biosynthesis
  • Enzymes, Immobilized / genetics
  • Enzymes, Immobilized / pharmacology
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Fibroblasts / cytology
  • Fibroblasts / drug effects
  • Gene Expression
  • Genetic Vectors / chemistry
  • Genetic Vectors / metabolism
  • Glycoside Hydrolases / biosynthesis
  • Glycoside Hydrolases / genetics
  • Glycoside Hydrolases / pharmacology*
  • Mice
  • Protein Domains
  • Pseudomonas aeruginosa / drug effects*
  • Pseudomonas aeruginosa / growth & development
  • Pseudomonas aeruginosa / pathogenicity
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / genetics
  • Recombinant Proteins / pharmacology

Substances

  • Anti-Bacterial Agents
  • Bacterial Proteins
  • Enzymes, Immobilized
  • Recombinant Proteins
  • Cellulose
  • Glycoside Hydrolases