One-pot preparation of laccase@yttrium phosphate hybrid nanostructures as a novel antibiofilm agent

BACKGROUND AND OBJECTIVESA number of strategies have been developed in the last decade to inhibit biofilm formation on biotic and abiotic surfaces. Today, laccases are known to stand as important antibiofilm agents because these molecules can inhibit biofilm formation in an eco-friendly manner. A detailed antibiofilm mechanism of laccase includes penetrating the microbial membrane and oxidizing essential proteins, enzymes, or other cellular sites. For biofilm removal, enzyme immobilization provides excellent stability, high reusability, and better efficiency. In this regard, the use of hybrid nanostructures (HNSs) for enzyme immobilization has gained significant attention due to the ease of preparation and enhanced catalytic activity. Herein, the novel hybrid nanoflowers consisting of yttrium (III) ions combined with laccase were prepared and their antibiofilm activity was evaluated against some pathogenic bacteria.MATERIALS AND METHODSThe hybrid particles were synthesized in an aqueous solution using yttrium phosphate (Y۳(PO۴)۲) as the inorganic component and laccase as the organic constituent. Then, the effects of reaction parameters on the formation of the enzyme-embedded hybrid nanoparticles were investigated. Furthermore, the antibiofilm activity of the prepared HNSs was evaluated against Pseudomonas aeruginosa, Escherichia coli, Bacillus subtilis, and Staphylococcus aureus. Finally, the biofilm formation in the presence of the free enzyme and laccase@YPO۴•HNP was visualized by crystal violet staining and Confocal Laser Scanning Microscopy (CLSM).RESULTS AND DISCUSSIONThe free laccase did not significantly prevent biofilm formation even increasing concentration up to ۱۲۰ U L–۱. Laccase@YPO۴•HNPs effectively inhibited E. coli, S. aureus, and B. subtilis biofilms at concentrations of ≤ ۶۰ U L–۱. According to the crystal violet staining and CLSM images, the total cell coverage for the biofilms significantly declined compared to the control following the treatment with laccase@YPO۴•HNPs.CONCLUSIONLaccase@YPO۴•HNPs could be considered a novel, eco-friendly, and antibiofilm heterogeneous biocatalyst for medicine and industry.

کلیدواژه ها:

Laccase ، Yttrium phosphate ، Antibiofilm ، Hybrid nanostructures