A Metabolomic Approach of Ai-Driven Enzymatic Digestion of Pumpkin Seed Flour for Producing Umami Metabolites

This study presents an innovative approach that combines enzymatic digestion with AI-driven optimization to produce pumpkin seed (Cucurbita pepo) metabolites that resemble those of chicken. Proteolytic enzymes, such as subtilisin, a mix of leucine aminopeptidase and glutamyl endopeptidase, and glutaminase, were used to break down the protein matrix and release key amino acids, with glutamate standing out for its umami-enhancing properties. Using AI-optimized reaction conditions reduces the number of trials needed and maximizes the production of desired metabolites. Metabolomic analyses revealed a substantial increase in compounds such as alanine and cysteine and the release of umami-boosting nucleotides (GMP and IMP). Sensory analyses showed that plant-based nuggets enriched with digested pumpkin seed flour exhibited flavor characteristics and acceptance levels comparable to traditional chicken nuggets. This approach highlights the potential of AI-optimized enzymatic technology to develop sustainable and appealing food alternatives, aligning with the growing demand for plant-based products that replicate the sensory experience of animal-based foods. © 2025 Elsevier B.V.