Assessment of Photoactivated Chlorophyllin Production of Singlet Oxygen and Inactivation of Foodborne Pathogens

Abstract

Singlet oxygen (1O2) is known to have antibacterial activity; however, production can involve complex processes with expensive chemical precursors and/or significant energy input. Recent studies have confirmed the generation of 1O2 through the activation of photosensitizer molecules (PSs) with visible light in the presence of oxygen. Given the increase in the incidence of foodborne diseases associated with cross-contamination in food-processing industries, which is becoming a major concern, food-safe additives, such as chlorophyllins, have been studied for their ability to act as PSs. The fluorescent probe Singlet Oxygen Sensor Green (SOSG®) was used to estimate 1O2 formation upon the irradiation of traditional PSs (rose bengal (RB), chlorin 6 (ce6)) and novel chlorophyllins, sodium magnesium (NaChl) and sodium copper (NaCuChl), with both simulated-solar and visible light. NaChl gave rise to a similar 1O2 production rate when compared to RB and ce6. Basic mixing was shown to introduce sufficient oxygen to the PS solutions, preventing the limitation of the 1O2 production rate. The NaChl-based inactivation of Gram-positive S. aureus and Gram-negative E. coli was demonstrated with a 5-log reduction with UV–Vis light. The NaChl-based inactivation of Gram-positive S. aureus was accomplished with a 2-log reduction after 105 min of visible-light irradiation and a 3-log reduction following 150 min of exposure from an initial viable bacterial concentration of 106 CFU mL−1. CHS-NaChl-based photosensitization under visible light enhanced Gram-negative E. coli inactivation and provided a strong bacteriostatic effect preventing E. coli proliferation. The difference in the ability of NaChl and CHS-NaChl complexes to inactivate Gram-positive and Gram-negative bacteria was confirmed to result from the cell wall structure, which impacted PS–bacteria attachment and therefore the production of localized singlet oxygen.