摘要
目的:以大肠杆菌表达的重组荞麦胰蛋白酶抑制剂为原材料,研究其固定化方法及条件。方法:以0.2%聚乙烯醇-3%海藻酸钠溶液为载体,CaCl2为固定剂,用物理包埋法对荞麦胰蛋白酶抑制剂进行固定化;在CaCl2浓度、载体与抑制剂体积比以及固定化时间3个单因素基础上,利用响应面法对荞麦胰蛋白酶抑制剂固定化的影响因素进行优化。结果:建立了响应面法优化固定荞麦胰蛋白酶抑制剂的模型,经优化后得到如下最佳固定化条件:CaCl2浓度为5.5%,载体与抑制剂体积比为1.6∶1,固定化时间为31min。在此条件下实际测得固定化抑制剂抑制率为72.4%,而模型预测此条件下的抑制率为74.3%,实测值与理论值相差很小。结论:所建模型拟合程度较高,用该模型优化荞麦胰蛋白酶抑制剂固定化的工艺条件参数准确可信,可为进一步开发胰蛋白酶的应用提供重要参考。
Objective: To study the immobilization conditions of buckwheat trypsin inhibitor(BTI) expressed by E.coli BL21 pExSec I-BTI. Methods: BTI was immobilized by physical entrapment with carriers of 0.2% polyvinyl alcohol-3% sodium alginate and fixative of CaCl 2 . Based on tests of single factor of CaCl 2 concentration, carri? er-inhibitor volume ratio and immobilization time, the response surface methodology was employed to optimize the overall immobilization conditions. Results: The optimal conditions of immobilize BTI were as follow: CaCl 2 concen? tration of 5.5%, the ratio of carrier to inhibitor volume of 1.6∶1, and the immobilization time of 31 min. Under these conditions, the optimum inhibitory activity of immobilized BTI was 72.4%, which was consistent with the predicted value of 74.3%. Conclusion: This response model showed the validity of optimizing the parameters for BTI immobilization, and it will be a reference for BTI development.
作者
李霞
栗安之
李晨
LI Xia;LI An-Zhi;LI Chen(School of Life Science, Shanxi University, Taiyuan 030006, China)
出处
《生物技术通讯》
CAS
2019年第3期391-396,413,共7页
Letters in Biotechnology
基金
国家自然科学基金青年科学基金(31600631)
山西省高校科技创新项目(2016116)
关键词
荞麦胰蛋白酶抑制剂
固定化
响应面法
buckwheat trypsin inhibitor
immobilization
response surface methodology
