Purification and Characterization of Transglutaminase Isolated from Sardine (Sardina pilchardus) Flesh Waste
Transglutaminase (TGase) is an enzyme that catalyzes acyl transfer reactions by creating covalent cross-links between protein molecules and has been used to improve the physical and functional properties of protein-based foods. The objectives of this study were the extraction, purification, and bioc...
| Autores: | , , , |
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| Formato: | artículo |
| Estado: | Versión publicada |
| Fecha de publicación: | 2025 |
| País: | España |
| Recursos: | Universidad de Sevilla (US) |
| Repositorio: | idUS. Depósito de Investigación de la Universidad de Sevilla |
| OAI Identifier: | oai:idus.us.es:11441/172519 |
| Acesso em linha: | https://hdl.handle.net/11441/172519 https://doi.org/10.3390/polym17040510 |
| Access Level: | acceso abierto |
| Palavra-chave: | Transglutaminase Sardine Enzyme purification Enzyme characterization Cross-linking Actomyosin |
| Resumo: | Transglutaminase (TGase) is an enzyme that catalyzes acyl transfer reactions by creating covalent cross-links between protein molecules and has been used to improve the physical and functional properties of protein-based foods. The objectives of this study were the extraction, purification, and biochemical characterization of TGase from sardine (Sardina pilchardus) flesh in order to provide a suitable TGase enzyme for food industry applications. The results showed a specific activity, yield, and purification fold of 357.14 U/mg protein, 36.74%, and 183.15, respectively. The enzyme exhibited maximal activity at 40 ◦C and pH 8.0, with a molecular weight of around 57 kDa. The effect of time on TGase thermal stability at 40 ◦C showed a gradual decrease in its catalytic activity during the incubation time until the enzyme was completely inactivated at 60 min. Additionally, the sardine TGase was found to be calcium-dependent. However, Mg2+ and Ba2+ ions were found to be effective in its activation to some extent and a total inhibition was shown by Zn2+ and Sr2+ ions. The TGase activity was affected markedly by NaCl and EDTA, and lost, respectively, about 80.7% and 36.49% from its activity by increasing the concentration (1.5 M NaCl and 20 mM EDTA). Based on the surface hydrophobicity and solubility results, the cross-linking of natural actomyosin mediated by TGase increased to a greater extent. The results revealed that sardine TGase possessed attractive qualities, making it a potential alternative to other TGase sources for food industry applications. |
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