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<p>This study developed a novel two-step enzymatic modification strategy involving papain hydrolysis followed by microbial transglutaminase (TGase) cross-linking to enhance the functionality of soy protein isolate (SPI) for high-moisture meat analogue (HMMA) production. By systematically optimizing the degree of hydrolysis (DH) and cross-linking conditions, it was found that moderate hydrolysis (DH = 3.56%) combined with TGase cross-linking (30 U/g) significantly improved the hydration and emulsifying properties of SPI, with the oil holding capacity (OHC), emulsifying activity index (EAI), and emulsion stability index (ESI) increased by 18.45%, 40.28%, and 20.93%, respectively. High-moisture extrusion of the modified SPI resulted in HMMA with a more ordered and anisotropic fibrous structure, as evidenced by a fibrous degree of up to 1.86, an anisotropy index of up to 1.68, as well as enhanced hardness and springiness, which increased by 49.50% and 26.90%. These enhancements can be attributed to the rearrangement and covalent cross-linking of protein molecules, as well as the increased content of ordered structures such as <i>β</i>-sheets and disulfide bonds (S–S bonds) and improved water immobilization. Furthermore, the in vitro protein digestibility increased from 81.80% in the control group to 92.00% in the enzymatic hydrolysis group and 87.10% in the enzymatic hydrolysis-cross-linking group, indicating superior nutritional quality. These findings demonstrate that the synergistic “enzymatic hydrolysis-cross-linking” strategy effectively restructured SPI, providing a promising approach for producing plant-based meat alternatives with meat-like texture and enhanced performance.</p> Graphical Abstract <p></p>

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Synergistic Enzymatic Strategy Using Papain and Transglutaminase to Engineer the Structure and Texture of High-Moisture Plant-Based Meat Analogs

  • 期刊论文

  • 发表日期：

    2026年6月3日

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  • Ning Wang,
  • Shanshan Liu,
  • Jie Yu,
  • Xiaoxuan Li,
  • Fuhui Qi,
  • Kaiyue Yuan,
  • Danqing Ye,
  • Wenjie Sui,
  • Tao Wu,
  • Min Zhang

摘要

This study developed a novel two-step enzymatic modification strategy involving papain hydrolysis followed by microbial transglutaminase (TGase) cross-linking to enhance the functionality of soy protein isolate (SPI) for high-moisture meat analogue (HMMA) production. By systematically optimizing the degree of hydrolysis (DH) and cross-linking conditions, it was found that moderate hydrolysis (DH = 3.56%) combined with TGase cross-linking (30 U/g) significantly improved the hydration and emulsifying properties of SPI, with the oil holding capacity (OHC), emulsifying activity index (EAI), and emulsion stability index (ESI) increased by 18.45%, 40.28%, and 20.93%, respectively. High-moisture extrusion of the modified SPI resulted in HMMA with a more ordered and anisotropic fibrous structure, as evidenced by a fibrous degree of up to 1.86, an anisotropy index of up to 1.68, as well as enhanced hardness and springiness, which increased by 49.50% and 26.90%. These enhancements can be attributed to the rearrangement and covalent cross-linking of protein molecules, as well as the increased content of ordered structures such as β-sheets and disulfide bonds (S–S bonds) and improved water immobilization. Furthermore, the in vitro protein digestibility increased from 81.80% in the control group to 92.00% in the enzymatic hydrolysis group and 87.10% in the enzymatic hydrolysis-cross-linking group, indicating superior nutritional quality. These findings demonstrate that the synergistic “enzymatic hydrolysis-cross-linking” strategy effectively restructured SPI, providing a promising approach for producing plant-based meat alternatives with meat-like texture and enhanced performance.

Graphical Abstract