The Pel Polysaccharide Can Serve a Structural and Protective Role in the Biofilm Matrix of Pseudomonas aeruginosa

Abstract

Bacterial extracellular polysaccharides are a key constituent of the extracellular matrix material of biofilms. Pseudomonas aeruginosa is a model organism for biofilm studies and produces three extracellular polysaccharides that have been implicated in biofilm development, alginate, Psl and Pel. Significant work has been conducted on the roles of alginate and Psl in biofilm development, however we know little regarding Pel. In this study, we demonstrate that Pel can serve two functions in biofilms. Using a novel assay involving optical tweezers, we demonstrate that Pel is crucial for maintaining cell-to-cell interactions in a PA14 biofilm, serving as a primary structural scaffold for the community. Deletion of pelB resulted in a severe biofilm deficiency. Interestingly, this effect is strain-specific. Loss of Pel production in the laboratory strain PAO1 resulted in no difference in attachment or biofilm development; instead Psl proved to be the primary structural polysaccharide for biofilm maturity. Furthermore, we demonstrate that Pel plays a second role by enhancing resistance to aminoglycoside antibiotics. This protection occurs only in biofilm populations. We show that expression of the pel gene cluster and PelF protein levels are enhanced during biofilm growth compared to liquid cultures. Thus, we propose that Pel is capable of playing both a structural and a protective role in P. aeruginosa biofilms. Most bacteria live within biofilm communities, which are a complex population of microorganisms that attach to surfaces and produce copious amounts of extracellular matrix material. Exopolysaccharides are a key feature of the extracellular matrix and are found in many forms, ranging from structurally simple linear homopolymers to structurally complex branched heteropolymers. Exopolysaccharides carry out a wide range of functions involving adherence to surfaces and other cells, structural support and protection against host and environmental stress. The goal of our study was to examine the functional importance of polysaccharide production in the model biofilm organism, Pseudomonas aeruginosa. Using a deletion and over expression strategy, we characterized the function of one polysaccharide, Pel, and demonstrated that this polysaccharide has two roles, a structural role and a protective role, against an important class of antibiotics, aminioglycosides.