重组猪羧肽酶B的酶学性质及稳定性研究.PDFVIP

食品与药品 Food and Drug 2012年第 14卷第05期 159 重组猪羧肽酶B 的酶学性质及稳定性研究 吴 珊，李素霞* （华东理工大学生物反应器工程国家重点实验室，上海200237 ） 摘 要：目的 研究重组猪羧肽酶B 的酶学性质及稳定性。方法 测定其酶学动力学参数K 和V ，最适pH和最适温度；研 m max 究不同pH和不同温度条件下酶的稳定性，以及金属离子和EDTA对酶的活性与稳定性的影响。结果 重组猪羧肽酶B 的Km 为 0.27 mmol/L ，V 为222.22 μmol/min ，最适pH为7.65，最适催化温度为25 ℃；pH在5.0~9.0范围内，温度在4~30 ℃稳定性 max 良好，Cu2+和EDTA抑制酶活性，Co2+促进酶活性。结论 重组猪羧肽酶B 的酶学性质与报道的提取猪羧肽酶B 的性质基本相 同，pH5.0~9.0范围内稳定，在4~30 ℃时稳定性好。 关键词：重组猪羧肽酶B ；酶学性质；稳定性 中图分类号：Q555 文献标识码：A 文章编号：1672-979X(2012)05-0159-04 Study on Property and Stability of Recombinant Porcine Carboxypeptidase B in E. coli WU Shan, LI Su-xia (State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China) Abstract: Objective To research the enzymatic property and stability of recombinant porcine pancreatic carboxypeptidase B (CPB). Methods The kinetic parameters K and V , optimum pH and temperature were m max measured; the effects of pH and temperature on stability of CPB were studied, the effects of different metal ions and EDTA on activity and stability of CPB were also investigated. Results The K and V measured were 0.27 mmol/ m max L and 222.22 μmol/ min, respectively. The optimum pH was 7.65, the optimum catalytic temperature was 25℃. CPB was stable when pH was between 5.0 and 9.0 or temperature was lower than 30 ℃. Cu2+ and EDTA inhibited the activity of CPB while Co2+ promoted it. Conclusion The enzymatic property of recombinant