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Exploring the Milk-Clotting and Proteolytic Activities in Different Tissues of Vallesia glabra: a New Source of Plant Proteolytic Enzymes

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Abstract

Proteolytic enzymes are widely distributed in nature, playing essential roles in important biological functions. Recently, the use of plant proteases at the industrial level has mainly increased in the food industry (e.g., cheesemaking, meat tenderizing, and protein hydrolysate production). Current technological and scientific advances in the detection and characterization of proteolytic enzymes have encouraged the search for new natural sources. Thus, this work aimed to explore the milk-clotting and proteolytic properties of different tissues of Vallesia glabra. Aqueous extracts from the leaves, fruits, and seeds of V. glabra presented different protein profiles, proteolytic activity, and milk-clotting activity. The milk-clotting activity increased with temperature (30–65 °C), but this activity was higher in leaf (0.20 MCU/mL) compared with that in fruit and seed extracts (0.12 and 0.11 MCU/mL, respectively) at 50 °C. Proteolytic activity in the extracts assayed at different pH (2.5–12.0) suggested the presence of different types of active proteases, with maximum activity at acidic conditions (4.0–4.5). Inhibitory studies indicated that major activity in V. glabra extracts is related to cysteine proteases; however, the presence of serine, aspartic, and metalloproteases was also evident. The hydrolytic profile of caseins indicated that V. glabra leaves could be used as a rennet substitute in cheesemaking, representing a new and promising source of proteolytic enzymes.

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Acknowledgments

Gonzalez-Velazquez wants to thank the National Council of Science and Technology of Mexico (CONACyT) for the economic support during postgraduate studies.

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Authors and Affiliations

  1. Coordinación de Tecnología de Alimentos de Origen Animal, Laboratorio de Química de Alimentos y Biotecnología de Productos Lácteos, Centro de Investigación en Alimentación y Desarrollo, A.C, Carretera Gustavo Enrique Astiazarán Rosas No. 46, Colonia La Victoria, CP. 83304, Hermosillo, Sonora, Mexico

    Daniel A. González-Velázquez, Miguel A. Mazorra-Manzano, Belinda Vallejo-Córdoba & Wendy G. Mora-Cortes

  2. Coordinación de Tecnología de Alimentos de Origen Animal, Laboratorio de Biología de Organismos Acuáticos, Centro de Investigación en Alimentación y Desarrollo, A.C., Gustavo Enrique Astiazarán Rosas No. 46, Colonia La Victoria, CP. 83304, Hermosillo, Sonora, Mexico

    Marcel Martínez-Porchas

  3. Laboratorio de Bioquímica de Proteínas y Glicanos. Carretera, CONACYT-Centro de Investigación en Alimentación y Desarrollo A.C., Gustavo Enrique Astiazarán Rosas No. 46, Colonia La Victoria, CP. 83304, Hermosillo, Sonora, Mexico

    José A. Huerta-Ocampo

  4. Instituto Nacional de Investigaciones Forestales, Agrícolas y Pecuarias (INIFAP), Carretera Culiacán-Eldorado km 17,5, Culiacán, Sinaloa, Mexico

    Jesús M. Moreno-Hernández

  5. Coordinación de Tecnología de Alimentos de Origen Animal, Laboratorio de Bioquímica y Calidad de Productos Pesqueros, Centro de Investigación en Alimentación y Desarrollo, A.C, Carretera. Gustavo Enrique Astiazarán Rosas No. 46, Colonia La Victoria, C.P. 83304, Hermosillo, Sonora, Mexico

    Juan C. Ramírez-Suarez

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  1. Daniel A. González-Velázquez
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  2. Miguel A. Mazorra-Manzano
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Correspondence to Miguel A. Mazorra-Manzano.

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González-Velázquez, D.A., Mazorra-Manzano, M.A., Martínez-Porchas, M. et al. Exploring the Milk-Clotting and Proteolytic Activities in Different Tissues of Vallesia glabra: a New Source of Plant Proteolytic Enzymes. Appl Biochem Biotechnol 193, 389–404 (2021). https://doi.org/10.1007/s12010-020-03432-5

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