Influence of protease type and concentration on protein digestibility, physicochemical and textural properties of extruded high-moisture meat analogues

Influence of protease type and concentration on protein digestibility, physicochemical and textural properties of extruded high-moisture meat analogues

Food Hydrocolloids

10.1016/j.foodhyd.2025.111910

https://doi.org/10.1016/j.foodhyd.2025.111910

Grace Cui Fang Ng, Caleb ONG, Alicia Hui Ping Theng, Dayna Shu Min Ong, Chaoyu DONG, Hui Yu Sim, Xin Yi Hua, Felicia Siew Kay Ng, Jie Hong Chiang

high-moisture meat analogues, extrusion, proteolytic enzyme, in-vitro digestion, Amino acids, molecular weight distributions, Texture

26 August 2025

Fang NG, G. C., ONG, C., Ping THENG, A. H., Min ONG, D. S., DONG, C., Yu SIM, H., Yi HUA, X., Kay NG, F. S., & Hong CHIANG, J. (2025). Influence of protease type and concentration on protein digestibility, physicochemical and textural properties of extruded high-moisture meat analogues. Food Hydrocolloids, 111910. https://doi.org/10.1016/j.foodhyd.2025.111910

High-moisture extrusion of plant proteins can produce meat analogues with fibrous structures and improved nutritional profiles, but replicating the meat-like texture and optimising protein digestibility remains challenging. This study investigated the effects of three proteolytic enzymes, Alcalase® (AL), Protana® Prime (PP), and Flavourzyme® (FL), added at varying concentrations (0, 0.5, 1, 2.5 and 5% w/w) on soy–pea protein-based meat analogues produced through high-moisture co-extrusion. Physicochemical and textural properties (hardness, chewiness, springiness, and degree of texturisation) were assessed alongside protein solubility, molecular weight distribution, and in vitro digestibility. Results indicated that low-to-moderate enzyme concentrations (0.5–1% w/w) slightly reduced hardness and chewiness while improving the fibrous structure, particularly in samples treated with FL and PP. In contrast, higher enzyme concentrations, especially with AL, induced extensive hydrolysis, resulting in fragmented extrudates and a diminished textural quality. Analysis of gastric and intestinal digesta revealed that all enzyme treatments increased soluble protein levels and the release of free amino acids compared to untreated controls. However, the specific profiles depended on enzyme selectivity. Notably, PP tended to generate higher amounts of free arginine, while AL excelled at releasing smaller peptide fragments. These findings highlight the potential to tailor textural attributes and nutritional quality in soy–pea meat analogues by selecting specific enzymes and controlling their addition, thereby paving the way for more meat-like products with enhanced protein digestibility and amino acid bioavailability.

Publisher Copyright

This research / project is supported by the A*STAR BMRC (Biomedical Research Council) - Singapore Food Story 2 Industry Alignment Fund - Pre-Positioning Future Foods: Alternative Proteins
Grant Reference no. : H20H8a0002

© 2025 Elsevier B.V. This is an Accepted Manuscript of an article published by Elsevier in Food Hydrocolloids, available at 10.1016/j.foodhyd.2025.111910.

https://oar.a-star.edu.sg/communities-collections/articles/22150

0268-005X

Singapore Institute of Food and Biotechnology Innovation