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J Agric Food Chem ; 72(15): 8742-8748, 2024 Apr 17.
Artículo en Inglés | MEDLINE | ID: mdl-38564658

RESUMEN

Tyrosinase is capable of oxidizing tyrosine residues in proteins, leading to intermolecular protein cross-linking, which could modify the protein network of food and improve the texture of food. To obtain the recombinant tyrosinase with microbial cell factory instead of isolation tyrosinase from the mushroom Agaricus bisporus, a TYR expression cassette was constructed in this study. The expression cassette was electroporated into Trichoderma reesei Rut-C30 and integrated into its genome, resulting in a recombinant strain C30-TYR. After induction with microcrystalline cellulose for 7 days, recombinant tyrosinase could be successfully expressed and secreted by C30-TYR, corresponding to approximately 2.16 g/L tyrosinase in shake-flask cultures. The recombinant TYR was purified by ammonium sulfate precipitation and gel filtration, and the biological activity of purified TYR was 45.6 U/mL. The purified TYR could catalyze the cross-linking of glycinin, and the emulsion stability index of TYR-treated glycinin emulsion was increased by 30.6% compared with the untreated one. The cross-linking of soy glycinin by TYR resulted in altered properties of oil-in-water emulsions compared to emulsions stabilized by native glycinin. Therefore, cross-linking with this recombinant tyrosinase is a feasible approach to improve the properties of protein-stabilized emulsions and gels.


Asunto(s)
Reactivos de Enlaces Cruzados , Expresión Génica , Globulinas , Hypocreales , Monofenol Monooxigenasa , Proteínas Recombinantes , Proteínas de Soja , Monofenol Monooxigenasa/biosíntesis , Monofenol Monooxigenasa/genética , Monofenol Monooxigenasa/aislamiento & purificación , Monofenol Monooxigenasa/metabolismo , Reactivos de Enlaces Cruzados/aislamiento & purificación , Reactivos de Enlaces Cruzados/metabolismo , Hypocreales/clasificación , Hypocreales/genética , Hypocreales/crecimiento & desarrollo , Hypocreales/metabolismo , Globulinas/química , Globulinas/metabolismo , Proteínas de Soja/química , Proteínas de Soja/metabolismo , Electroporación , Celulosa , Sulfato de Amonio , Cromatografía en Gel , Precipitación Fraccionada , Emulsiones/química , Emulsiones/metabolismo , Proteínas Recombinantes/biosíntesis , Proteínas Recombinantes/genética , Proteínas Recombinantes/aislamiento & purificación , Proteínas Recombinantes/metabolismo , Estabilidad Proteica , Retículo Endoplásmico/metabolismo , Señales de Clasificación de Proteína , Aceites/química , Agua/química
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