Recyclable metal nanoparticle-immobilized polymer dot on montmorillonite for alkaline phosphatase-based colorimetric...

Development of simultaneous bacteria detection and eradication with simple, rapid, and reusable material is important in addressing bacterial contamination issues. In this study, we utilized the expression of alkaline phosphatase (ALP) from bacteria to design fluorescence ON/OFF system for bacteria detection, also using metal oxide nanoparticle for obtaining antibacterial activity and recyclability. The fluorescent-based biosensor with antibacterial activity was prepared by intercalating ALP-sensitive polymer dot (PD) containing β-cyclodextrin (β-CD) onto montmorillonite (MMT) as loading matrix via ionic exchange reaction, followed by immobilization of magnetic iron oxide (Fe₃O₄) and NIR-responsive cesium tungsten oxide (CsWO₃). The PD-βCD-MMT/Fe₃O₄-CsWO₃ nanocomposite exhibited strong fluorescence intensity, which was quenched in the presence of bacterial ALP (0-1000 U/L) due to hydrolysis of p-nitrophenyl phosphate (NPP) into p-nitrophenol (NP) in the hydrophobic site of β-CD. Furthermore, the nanocomposite could detect both gram-negative Escherichia coli and gram-positive Staphylococcus aureus in the range of 10¹-10⁷ CFU/mL (LOD 5.09 and 4.62 CFU/mL, respectively), and showed high antibacterial activity against bacteria by generating photothermal heat under 5 min NIR irradiation, causing damage to bacterial cells. This material also demonstrated recyclability via magnetic field exposure due to the presence of Fe₃O_4. In addition, the fluorescence can be recovered following pH shock and re-conjugation of β-CD molecules. After 4 cycles, nanocomposite still showed stable photothermal effects and fluorescence-based bacteria detection. Thus, this reusable material offers promising approach for simultaneous bacteria detection and killing, which is simple, rapid, and effective.