The protease \u3b1-chymotrypsin (\u3b1-CT) was covalently immobilized on a low-density polyethylene (LDPE) surface, providing a new non-leaching material (LDPE-\u3b1-CT) able to preserve surfaces from biofilm growth over a long working timescale. The immobilized enzyme showed a transesterification activity of 1.24 nmol/h, confirming that the immobilization protocol did not negatively affect \u3b1-CT activity. Plate count viability assays, as well as confocal laser scanner microscopy (CLSM) analysis, showed that LDPE-\u3b1-CT significantly impacts Escherichia coli biofilm formation by (i) reducing the number of adhered cells ( 1270.7 \ub1 5.0%); (ii) significantly affecting biofilm thickness ( 1281.8 \ub1 16.7%), roughness ( 1213.8 \ub1 2.8%), substratum coverage ( 1263.1 \ub1 1.8%), and surface to bio-volume ratio (+7.1 \ub1 0.2-fold); and (iii) decreasing the matrix polysaccharide bio-volume (80.2 \ub1 23.2%). Additionally, CLSM images showed a destabilized biofilm with many cells dispersing from it. Notably, biofilm stained for live and dead cells confirmed that the reduction in the biomass was achieved by a mechanism that did not affect bacterial viability, reducing the chances for the evolution of resistant strains
