Antifungal Effect of the Proteolytic Fraction P1G10 Stabilized by Alginate-Chitosan Polyelectrolyte Complexation...

Botrytis cinerea is a major phytopathogen responsible for significant postharvest losses in plant-derived foods. The increasing resistance to synthetic fungicides has driven the search for sustainable alternatives, including enzyme-based biofungicides. In this study, the proteolytic fraction P1G10 from Vasconcellea pubescens latex was encapsulated in an alginate-chitosan (ALG-CS) matrix to improve its stability and antifungal performance. The encapsulated formulation (ALG-CS-P1G10) retained ~95% enzymatic activity after 8 h under stress conditions (37 °C, 1350 lux), compared with 67% for the free enzyme. In vitro assays demonstrated a dose-dependent inhibition of B. cinerea growth, with an IC₅₀ value of ~11 mg/mL determined using a logistic model. At this concentration, the formulation reduced fungal adhesion by more than 80% and increased sensitivity to cell wall-disrupting agents (Congo Red and Calcofluor White), pointing to alterations in cell wall integrity. Importantly, the encapsulated system provided a more stable and sustained antifungal effect, consistent with a controlled-release mechanism. These results demonstrate that coupling enzyme stabilization with controlled release can improve the functional performance of protease-based antifungal systems, offering a promising strategy for the development of biofungicides in postharvest applications.