摘要
用硫酸铵分级沉淀法从木瓜粉提取液中分离木瓜蛋白酶，将其偶联在对氨基苯磺酰乙基（ＡＢＳＥ）－交联琼脂上，制得固定化木瓜蛋白酶。实验结果表明：木瓜蛋白酶与ＡＢＳＥ－交联琼脂的最佳结合量是每克干交联琼脂１４．０μｍｏｌ／ｓ，固定化酶的最适反应ｐＨ为８．０，最适反应温度为７５℃，Ｋｍ（ａｐｐ）为６７．６ｍｍｏｌ／Ｌ。固定化酶对尿素的稳定性较高，用６ｍｍｏｌ／Ｌ脲处理过夜能保存８２％的活力，而游离酶在同样条件下将失去全部活性。
Papain was purified by precipitation of (NH 4) 2SO 4,and then immobilized on p aminobenzene sulphonyl ethyl(ABSE) crosslinked agarose. The support had a binding capacity of 14.0μmol/s.g dry agarose. The pH and temperature optima were 8.0 and 75℃ for immobilized paptain, and K m(app) was 67.6mmol/L. When treated with 6mol/L urea for all night, immobilized papain rest 82% of its activity, whereas soluble papain lost their activity under similar conditions.
出处
《华东理工大学学报（自然科学版）》
CAS
CSCD
北大核心
1997年第1期49-52,共4页
Journal of East China University of Science and Technology
关键词
木瓜蛋白酶
固定化
蛋白酶
制备
papain
immobilization
optimal conditions
K m
protein denaturant
