Functional characterization of purified pear protease and its proteolytic activities with casein and myofibrillar proteins

Abstract

This study was performed to characterize pear protease proteolytic activity and investigate the use of pear protease as a meat tenderizer. Pear protease was purified and stabilized by 5% dextrin during lyophilization (dry) or concentration (liquid). Pear protease was further characterized with respect to pH, thermodynamics, and enzyme kinetics. Pear protease was stable at a pH range of 5-8 with an optimum pH of 6.5. From Arrhenius plots, liquid protease showed higher temperature dependency (23.49 kJ/mol) than dry protease (18.62 kJ/mol) due to its higher activation energy. The kcat/Km, catalytic efficiency of enzyme, was similar with 2.9 and 2.7 µM/min with dry and liquid proteases. Pear protease was evaluated for its proteolytic activities with casein and beef myofibrillar proteins by individually and combination with fig and kiwifruit proteases. These result indicated that pear and kiwifruit proteases could be complementary to be a desirable product for meat tenderization.

Enzyme activity and partial characterization of proteases obtained from Bromelia karatas fruit and compared with Bromelia pinguin proteases

“…The development of techniques and methods for the purification of proteases is important for many biotechnological advances because the protease isolation treatment defines the kinetic behavior of these enzymes in different processes such as meat tenderization, brewing, protein hydrolysates [9], cheese production [10], and the pharmaceutical industry [11]. In the protocols for isolation of plant proteases, several extraction solutions have been used: water, phosphate buffer, Tris-HCl buffer, and sodium phosphate buffer.…”

Application of Plant Proteases in Meat Tenderization: Recent Trends and Future Prospects

“…Combining both these had a synergistic effect [ 91 ]. Pear protease has been recently explored as an ideal meat tenderizer with the desired meat tenderization level rather than overaction leading to mushy spots due to over proteolysis as with bromelain and actinidin [ 92 ]. In plant protease extracted from four overmatured fruits, viz., kiwifruit, pear, apple, and grape, kiwifruit demonstrated potent protease activity (921 U) followed by grape (225.86 U) > pear (97.75 U) > apple (78.29 U).…”

“…Nam et al [ 92 ] explored the potential to use purified pear protease as a meat tenderizer and noted that pear protease and kiwi fruit protease have synergistic effects and could produce good quality meat products. Pear protease was stable at a wide pH range (5–8) with a catalytic efficiency of 2.9–2.7 μM/min.…”

“…Pear protease was stable at a wide pH range (5–8) with a catalytic efficiency of 2.9–2.7 μM/min. Pear protease stabilized in 5% dextrin during lyophilization demonstrated good proteolytic activity on casein and beef myofibrillar proteins [ 92 ]. A combination of the proteolytic enzyme of kiwifruit and pineapple and sous vide cooking was observed to demonstrate a significant reduction in the hardness of pork foreshank, and an increase in the L* and b* color value with kiwifruit enzyme was 60% more effective than pineapple protease and commercial neutrase [ 120 ].…”

Characteristics of Flavor Enhancer Based on Moringa Leaves and Tempeh Using Protease Enzymes From Sap of Papaya and Crown Flower

“…The results of this study showed that the hydrolysis process using mixed enzymes at a percentage of 3% addition can produce more glutamic acid compared to the use of a single enzyme. Similarly, Nam et al [41] explored the potential for using purified pear protease as a meat tenderizer and noted that the combination of pear protease and kiwi fruit protease has a synergistic effect and can produce good quality meat products. Research by Linberg et al [42] also showed that the combination of proteases resulted in an increase in soluble protein yield of about 15% compared to a single protease.…”