Exploring the Milk-Clotting and Proteolytic Activities in Different Tissues of Vallesia glabra : a New Source of Plant P
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Exploring the Milk-Clotting and Proteolytic Activities in Different Tissues of Vallesia glabra: a New Source of Plant Proteolytic Enzymes Daniel A. González-Velázquez 1 & Miguel A. Mazorra-Manzano 1 & Marcel Martínez-Porchas 2 & José A. Huerta-Ocampo 3 & Belinda Vallejo-Córdoba 1 & Wendy G. Mora-Cortes 1 & Jesús M. Moreno-Hernández 4 & Juan C. Ramírez-Suarez 5 Received: 12 August 2020 / Accepted: 28 September 2020/ # Springer Science+Business Media, LLC, part of Springer Nature 2020
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. Keywords Vallesia glabra . Proteolytic enzymes . Plant tissues . Milk-clotting . Zymography
* Miguel A. Mazorra-Manzano [email protected] Extended author information available on the last page of the article
Applied Biochemistry and Biotechnology
Introduction Proteases are a group of enzymes that hydrolyze the peptide bonds of proteins, causing the release of polypeptides or free amino acids. They are classified based on their biochemical and structural characteristics into 62 clans divided into 268 families [1]. Plant proteolytic enzymes possess excellent catalytic properties and are present in different cellular compartments (e.g., mitochondria, chloroplasts, Golgi apparatus, and peroxisomes), playing fundamental roles in protein processing, degradation, assimilation, and replacement as well as in gene regulation, expression, and maturation of zymogens during plant development [2, 3]. Currently, proteases represent nearly 60% of the industrial enzymes marketed worldwide due to their attractive characteristics of specificity and stability during their use in
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