Synergistic Antimicrobial Activity by Light or Thermal Treatment and Lauric Arginate: Membrane Damage and Oxidative Stress.

The need for more effective antimicrobials is critical for the food industry to improve food safety and reduce spoilage of minimally processed foods. The present study was initiated to develop an efficient and novel antimicrobial approach which combines physical treatments (UV-A or mild heat) and generally recognized as safe lauroyl arginate ethyl (LAE) to inactivate surrogate strains, including and Synergistic inactivation of bacteria resulted in an ∼6-log reduction of target bacteria, while individual treatments resulted in <1.5-log inactivation under the same set of conditions. In addition, the synergistic mechanism between LAE and UV-A/mild heat was evaluated by supplementing with a variety of antioxidants for suppressing oxidative stress and measurement of cell membrane damage by nucleic acid release. These results demonstrate that the synergistic antimicrobial activity of LAE and mild physical stresses was suppressed by supplementation with antioxidants. The research also compared LAE with another membrane-targeting lipopeptide antimicrobial agent, polymyxin B, to understand the uniqueness of LAE-induced synergy. Briefly, differences in modes of action between LAE and polymyxin B were characterized by comparing the MIC, damage to liposomes, and oxidative stress generation. These differences in the mode of action between LAE and polymyxin B suggested that both compounds target cell membrane but significantly differ in mechanisms, including membrane disruption and oxidative stress generation. Overall, this study illustrates synergistic antimicrobial activity of LAE with light or mild heat and indicates a novel oxidative stress pathway that enhances the activity of LAE beyond membrane damage. This study highlights an effective antimicrobial processing approach using a novel combination of lauroyl arginate ethyl (LAE) and two different physical treatments, light (UV-A) and mild heat. Both combinations demonstrated synergistic inactivation against a model Gram-negative bacterium or a Gram-positive bacterium or both by a >5-log reduction. Further mechanistic study revealed that oxidative stress is responsible for synergistic inactivation between LAE and UV-A, while both membrane damage and oxidative stress are responsible for the synergistic combination between LAE and mild heat. The mode of action of LAE was further compared to that of polymyxin B and analyzed using artificial membrane model systems and the addition of antioxidants. The proposed combination of LAE and common physical treatments may improve food preservation, food safety, and current sanitation processes for the food industry and the inactivation of pathogenic strains in biomedical environments.