Storage conditions and antiviral efficacy of yeast-derived vacuoles on T4 virus

In response to the urgent need for effective antiviral agents, this study explores the potential of vacuoles isolated from yeast in combating non-enveloped tailed viruses, using the T4 virus as a model. Concentration- and time-dependent assays revealed that vacuoles significantly inhibit T4 virus infectivity, achieving over 80% inhibition at 250 μg/mL. Morphological analysis via Bio-TEM imaging unveiled structural changes in the T4 virus after vacuole treatment, including separation of the capsid and tail, leading to impaired virion integrity. Optimization of vacuole storage conditions, particularly storing vacuoles in a pellet state, enhanced their antiviral efficiency. Characterization studies revealed structural modifications in vacuoles stored in the pellet state, such as increased particle size and changes in surface charge properties, potentially facilitating increased interaction with virion particles. These findings underline the promising potential of yeast-derived vacuoles as eco-friendly and effective antiviral agents against non-enveloped tailed viruses and provide insights into their mechanism of action. Further research is needed to elucidate molecular-level interactions and evaluate efficiency against other non-enveloped viruses. By offering novel insights into the antiviral potential of vacuoles, this study contributes to the development of eco-friendly antiviral strategies to address global health challenges. IMPORTANCE Non-enveloped viruses remain difficult to inactivate without harsh chemicals or heat. This study introduces yeast-derived vacuoles as a biologically based antiviral platform that disables a model non-enveloped bacteriophage (T4) with >80% inhibition at 250 μg/mL. Bio‑TEM reveals capsid-tail disassembly after vacuole exposure, linking macroscopic loss of infectivity to a defined structural mechanism. Storage engineering—maintaining vacuoles in pellet form—enhances efficacy and correlates with increased particle size and altered surface charge, suggesting tunable physicochemical interactions with virions. These results establish vacuoles as scalable, eco-friendly antiviral agents and provide design rules (dose, contact time, storage state, surface properties) for optimizing activity. Because the approach targets virion integrity rather than specific proteins, it may generalize across non-enveloped viruses, motivating molecular-level studies and translational testing. The work broadens the antiviral toolkit by leveraging a safe, low-cost cellular organelle.