Biofilm formation is a major cause of post-harvest microbial persistence on fresh produce, processing, and packaging surfaces, as the biofilm matrix has been shown to provide pathogens and other bacteria with protection from common disinfection approaches. The potential of an enzyme-based food processing aide has been investigated as a green, targeted supplement to common disinfection practices for preventing bacterial adhesion and removing mature biofilms. Crystal violet staining of biofilms formed in multi-well polystyrene plates has been used to demonstrate the efficacy of enzymatic biofilm prevention and removal on E. coli O157:H7, E. coli 25922, Salmonella Typhimurium, and Listeria monocytogenes. In the presence of 0.1 mg/mL enzyme, biofilm development was significantly inhibited for all bacteria, with a maximum of 41 ± 7% reduction for E. coli O157:H7. Effectiveness of mature biofilm removal varied by bacteria species type, with a maximum of 35 ± 12% reduction for E. coli O157:H7. A microfluidic flow cell was used to directly observe and quantify the impact of enzyme rinses on E. coli O157:H7 cells adhered to spinach leaf surfaces. In the flow cell, enzyme rinses resulted in significantly greater cell removal than water, representing a reversal of initial phases of biofilm formation. Initial biochemical chracterization of the enzyme indicates that it may degrade extracellular polysaccharides and other cell surface polymers, which can play an important role in bacterial adhesion and biofilm formation. Transmission electron microscopy (TEM) of treated and untreated cells reveal major modification to surface polysaccharide structures, consistent with the predicted mechanism of enzyme action. Additionally, changes in cell surface hydrophobicity were measured using the microbial adhesion to hydrocarbons (MATH) test and provide further evidence of changes to the cell surface. These results present a strong case for further development and optimization of enzyme activity to be applied as a novel food processing and packaging aid to minimize food safety risks associated with bacterial pathogens.

Reducing foodborne bacterial biofilms on produce and packaging with an engineered enzyme

Macromolecular Chemistry: The Second Century

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ACS Spring 2020 National Meeting & Expo

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.