Bacterial colonization and biofilm formation on surfaces are typically mediated by the deposition of exopolysaccharides and conditioning protein layers. Pseudomonas aeruginosa is a nosocomial opportunistic pathogen that utilizes strain-specific exopolysaccharides such as Psl, Pel or alginate for both initial surface attachment and biofilm formation. To generate surfaces that resist P. aeruginosa colonization, we covalently bound a Psl-specific glycoside hydrolase (PslG_h) to several, chemically-distinct surfaces using amine functionalization (APTMS) and glutaraldehyde (GDA) linking. In situ quartz crystal microbalance (QCM) experiments and confocal microscopy demonstrated a complete lack of Psl adsorption on the PslG_h-bound surfaces. Covalently-bound PslG_h was also found to significantly reduce P. aeruginosa surface attachment and biofilm formation over extended growth periods (8 days). The PslG_h surfaces showed a ∼99.9% (∼3-log) reduction in surface associated bacteria compared to control (untreated) surfaces, or those treated with inactive enzyme. This work demonstrates a non-eluting ‘bioactive’ surface that specifically targets a mechanism of cell adhesion, and that surface-bound glycoside hydrolase can significantly reduce surface colonization of bacteria through local, continuous enzymatic degradation of exopolysaccharide (Psl). These results have significant implications for the surface design of medical devices to keep bacteria in a planktonic state, and therefore susceptible to antibiotics and antimicrobials.

表面上的细菌定植和生物膜形成通常由胞外多糖的沉积和调节蛋白层介导。铜绿假单胞菌是一种医院机会病原体，它利用菌株特异性胞外多糖（例如Psl，Pel或藻酸盐）进行初始表面附着和生物膜形成。为了生成抗铜绿假单胞菌定植的表面，我们使用胺官能化（APTMS）和戊二醛（GDA）连接将Psl特异性糖苷水解酶（PslG _h）共价结合到几个化学上不同的表面。原位石英晶体微量天平（QCM）实验和共聚焦显微镜表明，PslG _h上完全缺乏Psl吸附绑定的表面。还发现共价结合的PslG _h在延长的生长期（8天）内可显着减少铜绿假单胞菌的表面附着和生物膜形成。PslG _h与对照（未处理）表面或未用酶处理的表面相比，与表面相关的细菌减少了约99.9％（约3-log）。这项工作表明非洗脱的“生物活性”表面特别针对细胞粘附机制，并且表面结合的糖苷水解酶可以通过胞外多糖（Psl）的局部连续酶促降解显着减少细菌的表面定殖。这些结果对于使细菌保持浮游状态并因此易受抗生素和抗菌剂影响的医疗设备的表面设计具有重要意义。