Abstract
Bacterial biofilms are a complex architecture of cells that grow on moist interfaces, and are held together by a molecular glue of extracellular proteins, sugars and nucleic acids. Biofilms are particularly problematic in human healthcare as they can coat medical implants and are thus a potential source of disease. The enzymatic dispersal of biofilms is increasingly being developed as a new strategy to treat this problem. Here, we have characterized NucB, a biofilm-dispersing nuclease from a marine strain of Bacillus licheniformis, and present its crystal structure together with the biochemistry and a mutational analysis required to confirm its active site. Taken together, these data support the categorization of NucB into a unique subfamily of the ββα metal-dependent non-specific endonucleases. Understanding the structure and function of NucB will facilitate its future development into an anti-biofilm therapeutic agent.
Highlights
Free-living, motile bacteria can develop into a stationary, multicellular community of cells known as a biofilm, a colony of sessile cells that forms on natural or artificial moist surfaces [1,2]
Medical implants and devices can be contaminated by biofilms [3], and dental caries [4], upper respiratory tract [5], ear infections [6] and chronic lung infections in cystic fibrosis patients are caused by biofilms [7,8]
To shed light on the biofilm-dispersing properties of NucB, we present here an analysis of the mode of action of this novel nuclease, its crystal structure and a mutational analysis that confirms the active site
Summary
Free-living, motile bacteria can develop into a stationary, multicellular community of cells known as a biofilm, a colony of sessile cells that forms on natural or artificial moist surfaces [1,2]. EDNA is a critical component of the ECM [18,19,20] and is required for the initial adhesion phase [21]. Secreted proteases [16] and glycoside hydrolases [17] degrade the protein and carbohydrate components of the ECM, respectively [15]. It was first shown several decades ago that biofilms treated with bovine DNase I had reduced viscosity [22] leading to biofilm dispersal [21]. There is increasing evidence that secreted nucleases play important roles in biofilm formation, dispersal and remodelling in many bacterial phyla, including major pathogens of humans such as Pseudomonas aeruginosa [7,8], Vibrio cholera [23] and Staphylococcus aureus [19]
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