Three phase partitioning to concentrate milk clotting proteases from Wrightia tinctoria R. Br and its characterization

Biological protein precipitation: A green process for the extraction of cucumisin from melon (Cucumis melo L. inodorus) by-products

“…The ample range of temperatures showed the high thermal stability of both MPJ and COM, producing the best MCA values at 85 • C with titles of 6300 and 4200 MCU/mg protein, respectively. These results are in concordance with the reported for milk-clotting proteases from Citrus aurantium L., Wrightia tinctoria and Morinda citrifolia L. at temperatures of 35-80 • C (de Farias et al, 2020;Mazorra-Manzano et al, 2013;Rajagopalan & Sukumaran, 2018). To the best of our knowledge, these findings were obtained for the first time from melon by-products, suggesting that MPJ still maintained bioactive proteins with relevant enzymatic activity, which can be effectively extracted by the natural chemical interaction with CRG without any previous purification step, leading a reduction of process time and cost when compare with traditional extractive methods.…”

“…This decreases the net charge, and therefore, decreases the electrostatic repulsion between charged groups, leading to milk clots agglomeration and enzymatic action (Ranadheera et al, 2019). Previous reports on MCA from plant proteases (Wrightia tinctorial, Ficus carica, Colatropis procera) also exhibited activity over a wide range of pH (Rajagopalan & Sukumaran, 2018). Fig.…”

Structural Analysis of a Novel Aspartic-Type Endopeptidase from Moringa oleifera Seeds and Its Milk-Clotting Properties

“…This was consistent with the observation that both the extracellular metalloproteinases (ACPs) isolated from Termitomyces clypeatus MTCC 5091 by Majumder et al and the protease isolated from Wrightia tinctoria rhizome by Rajagopalan et al preferentially hydrolyzed κ-CN. 24,25 In the positive control group (to the left of lane M in Figure 2A−C), calf rennet also hydrolyzed the three caseins to various degrees; α-CN hydrolysis did not occur within 5 min, while β-CN and κ-CN showed a certain degree of hydrolysis and an obvious product band appeared near the band of κ-CN, indicating that calf rennet also first hydrolyzed κ-CN and produced hydrolysates. This is consistent with the hydrolysis specificity of calf rennet previously reported by Uniacke-Lowe et al 26 and Jensen et al 27 Furthermore, the original bands corresponding to α-CN and β-CN did not change after 1 h, although the κ-CN band weakened significantly at 3 h. This indicated that the aspartic-type endopeptidase did not further hydrolyze α-CN and β-CN after 1 h, whereas it continued to hydrolyze κ-CN; this also implied that the extent of hydrolysis of κ-CN was higher than those of α-CN and β-CN.…”

Production of Plant Proteases and New Biotechnological Applications: An Updated Review

“…Once the material was conditioned, the solubilization process is performed, obtaining an extract with the target protease in a soluble form. The efficiency of the solubilization process and the distribution coefficient predominantly depend on material composition (protein, lipids, carbohydrates, pigments, fibers, polyphenols, gums, polysaccharides among other compounds), solvent buffer, extraction time, pH, ionic strength, reducing agents, and temperature [77] . During the solubilization process, rigorous control of operating conditions is required to prevent protein denaturation and functional adverse reactions.…”

“…The efficiency of the solubilization process and the distribution coefficient predominantly depend on material composition (protein, lipids, carbohydrates, pigments, fibers, polyphenols, gums, polysaccharides among other compounds), solvent buffer, extraction time, pH, ionic strength, reducing agents, and temperature. [77] During the solubilization process, rigorous control of operating conditions is required to prevent protein denaturation and functional adverse reactions. For most plant proteases, the extraction is carried out at pH between 4 and 9 because solubilization at extreme basic pH could produce the racemization of amino acids.…”