掌叶半夏胰蛋白酶抑制剂的分离纯化及其性质

掌叶半夏胰蛋白酶抑制剂的分离纯化及其性质 Purification and Characterization of a Trypsin Inhibitor with Insecticidal Activity fromPinellia pedatisecta 范汉东 FAN Handong1 1 2 first-author 郭建军 GUO Jianjun1 1 杨一兵 YANG Yibing1 1 邹国林 ZOU Goulin2 2 江西省科学院微生物研究所 江西省科学院微生物研究所 Institute of Microbiology,Jiangxi Academy of Sciences Institute of Microbiology,Jiangxi Academy of Sciences 武汉大学生命科学学院/病毒学国家重点实验室 武汉大学生命科学学院/病毒学国家重点实验室 College of Life Sciences,Wuhan University/State Key Laboratory of Virology College of Life Sciences,Wuhan University/State Key Laboratory of Virology 经硫酸铵分级沉淀、热处理、DEAE等方法从掌叶半夏中分离出一种不含糖的具有抗虫作用的胰蛋白酶抑制剂（Pinellia pedatisecta trypsin in hibitor,PPTI）,SDS-PAGE电泳与反相液相色谱（RT-HPLC）检测证明分离得到一纯蛋白,质谱（LC-MS）测定其相对分子质量为20596.09.荧光光谱分析研究结果显示:PPTI在pH低于9、温度低于60℃时,PPTI的三级结构相对稳定;抗虫活性结果显示有较好的抗芽虫作用. A trypsin inhibitor named Pinellia pedatisectatrypsin inhibior（PPTI）without sugar chain was isolated fromPinellia pedatisecta by heat treatment,salting out and DEAE ionic exchange chromatographic analysis.The purity was validated by SDS-PAGE as well as RT-HPLC.The relative molecular weight of PPTI was 20 596.09which was measured by LC-MS.PPTI was heat-tolerating when heated up to 60℃for 10min,at the same time,the tertiary structure was relatively stable when PPTI was dissolved in the buffer whose pH was lower than 9.Secticidal experiment showed it was effective against Aphids. 掌叶半夏 胰蛋白酶抑制剂 抗虫蛋白 Pinellia pedatisecta trypsin inhibitor insecticidal protein 10.14188/j.1671-8836.2010.05.016 S567.239 国家自然科学基金资助项目（30670464）;江西省科技支撑计划项目（2009BNA06600;2008BC34400）;江西省科学院国家级预研项目（2008-1-01） 2010-05-01 2021-04-01 5 56