RT Journal Article SR Electronic T1 Exopolysaccharide biosynthetic glycoside hydrolases can be utilized to disrupt and prevent Pseudomonas aeruginosa biofilms JF bioRxiv FD Cold Spring Harbor Laboratory SP 032714 DO 10.1101/032714 A1 Perrin Baker A1 Preston J. Hill A1 Brendan D. Snarr A1 Noor Alnabelseya A1 Mathew J. Pestrak A1 Mark J. Lee A1 Laura K. Jennings A1 John Tam A1 Roman Melnyk A1 Matthew R. Parsek A1 Donald C. Sheppard A1 Daniel J. Wozniak A1 P. Lynne Howell YR 2015 UL http://biorxiv.org/content/early/2015/11/26/032714.abstract AB Bacterial biofilms are a significant medical challenge as they are recalcitrant to current therapeutic regimes. A key component of biofilm formation in the opportunistic human pathogen Pseudomonas aeruginosa is the biosynthesis of the exopolysaccharides Pel and Psl, which are involved in the formation and maintenance of the structural biofilm scaffold and protection against antimicrobials and host defenses. Given that the glycoside hydrolases – PelAh and PslGh – encoded in the pel and psl biosynthetic operons, respectively, are utilized for in vivo exopolysaccharide processing, we reasoned that these would provide specificity to target P. aeruginosa biofilms. Evaluating these enzymes as potential therapeutics, we demonstrate that these glycoside hydrolases selectively target and degrade the exopolysaccharide component of the biofilm matrix and that nanomolar concentrations of these enzymes can both prevent biofilm formation as well as rapidly disrupt preexisting biofilms in vitro. This treatment was effective against clinical and environmental P. aeruginosa isolates and reduced biofilm biomass by 58–94%. These non-cytotoxic enzymes potentiated antibiotics as the addition of either enzyme to a sub-lethal concentration of colistin reduced viable bacterial counts by 2.5 orders of magnitude. Additionally, PelAh was able to increase neutrophil killing by ~50%. This work illustrates the feasibility and benefits of using bacterial exopolysaccharide biosynthetic glycoside hydrolases and synthetic biology to develop novel anti-biofilm therapeutics.