Pioneering EnzymMeat® Biotechnology: Direct Koji Application for Enhancing Physicochemical Quality, Sensory Performance, Satiety and Natural Bioprotection in fresh Beef

This work presents the first documented global study evaluating the direct application of Aspergillus oryzae (Koji) cellular mass and enzymatic suspensions on fresh beef, establishing the foundation of EnzymMeat® biotechnology. The study investigates the physicochemical behavior, sensory performance, satiety effects, and natural bioprotective mechanisms induced by Koji-based preparations applied to two anatomical regions of beef (round and neck). Koji treatments significantly improved moisture retention, reduced cooking losses, enhanced marinade uptake, and increased juiciness, tenderness, and flavor intensity. Variants activated at moderate temperatures (25 °C) demonstrated superior water-holding capacity, while salt-assisted preparations achieved the highest absorption. Sensory evaluation confirmed improved mouthfeel and overall preference, and satiety assessment indicated stronger and more persistent post-prandial fullness in Koji-treated samples. Physicochemical analysis showed moisture contents above 52–56% in Koji-treated beef, compared to <50% in controls, and cooking losses of 25–38% depending on activation temperature. pH values ranged from 4.60 to 5.16, reflecting moderate acidification driven by kojic, citric, and succinic acids. These mechanisms, combined with controlled proteolysis and the generation of bioactive peptides (<3 kDa), contributed to oxidative stability, antimicrobial protection, and extended refrigerated shelf-life. EnzymMeat® biotechnology operates through a synergistic set of natural processes: enzymatic proteolysis, acidification, bioactive peptide formation, antioxidant activity, and microbial competition. Together, these mechanisms position Koji as a clean-label, multifunctional fungal biotechnology capable of enhancing technological, sensory, nutritional, and safety attributes of fresh beef. This article provides a complete experimental framework, including sample preparation, physicochemical measurements, sensory evaluation, satiety assessment, and statistical analysis. The results establish EnzymMeat® as a pioneering platform in meat science and offer a reproducible methodology for future research and industrial applications.