纯化梨蛋白酶的功能特性及其对酪蛋白和肌原纤维蛋白的蛋白水解活性
Functional characterization of purified pear protease and its proteolytic activities with casein and myofibrillar proteins.
作者信息
Nam Seung-Hee, Kim Young-Min, Walsh Marie K, Yim Sun-Hee, Eun Jong-Bang
机构信息
1Institute of Agricultural Science and Technology and Department of Food Science & Technology, Chonnam National University, Gwangju, 61186 Korea.
2Department of Food Science & Technology and BK21 Plus Program, Graduate School of Chonnam National University, Gwangju, 61186 Korea.
出版信息
Food Sci Biotechnol. 2016 Mar 31;25(Suppl 1):31-39. doi: 10.1007/s10068-016-0095-6. eCollection 2016.
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.
本研究旨在表征梨蛋白酶的蛋白水解活性，并研究梨蛋白酶作为肉类嫩化剂的用途。在冻干（干粉）或浓缩（液体）过程中，梨蛋白酶用5%糊精进行纯化和稳定化处理。进一步对梨蛋白酶的pH值、热力学和酶动力学进行了表征。梨蛋白酶在pH值5-8的范围内稳定，最适pH值为6.5。根据阿伦尼乌斯曲线，由于液体蛋白酶具有较高的活化能，其显示出比干粉蛋白酶更高的温度依赖性（23.49 kJ/mol），而干粉蛋白酶为18.62 kJ/mol。酶的催化效率kcat/Km在干粉和液体蛋白酶中相似，分别为2.9和2.7 μM/min。通过单独以及与无花果蛋白酶和猕猴桃蛋白酶组合，评估了梨蛋白酶对酪蛋白和牛肉肌原纤维蛋白的蛋白水解活性。这些结果表明，梨蛋白酶和猕猴桃蛋白酶可以互补，成为肉类嫩化的理想产品。
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