Pea Protein Isolate—Enhanced Nanostructured Lipid Carriers with Beeswax for Limonene Encapsulation: Exploring Physicochemical Properties and In Vitro Insights

Abstract Nanostructured lipid carriers (NLCs) have emerged as a promising approach to enhance the stability and bioavailability of volatile bioactive compounds. This study introduces a novel pea protein isolate (PPI)‐stabilized NLC using beeswax as a solid lipid for limonene encapsulation, providing a plant‐based alternative to synthetic emulsifiers. The nanocarriers were prepared via melt emulsification, with an optimized ratio of beeswax‐to‐oleic acid to enhance stability and encapsulation efficiency (EE). Pea protein isolate acted as an emulsifier, enabling precise control over the particle size. The optimal formulation used a beeswax‐to‐oleic acid ratio of 3:1, with 3% pea protein isolate, a hydrodynamic diameter ranging from 120 nm to 155 nm, and a zeta potential below −40 mV, successfully encapsulating more than 70% of limonene. DSC analysis indicated alterations in the crystallinity of beeswax upon the addition of oleic acid, facilitating the incorporation of active compounds as nanostructured carriers. In vitro release studies demonstrated a controlled release profile, suggesting prolonged bioactive retention. These findings underscore the potential of pea protein isolate‐stabilized beeswax‐based NLCs as an innovative and sustainable platform for essential oil encapsulation. Additionally, these findings have promising applications in food preservation, pharmaceuticals, and nutraceuticals, offering a new avenue for enhancing the stability and bioavailability of the bioactive compounds.