...
  • 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Journal of Agricultural and Food Chemistry >Bowman-Birk Proteinase Inhibitor from Cajanus cajan Seeds: Purification, Characterization, and Insecticidal Properties
【24h】

Bowman-Birk Proteinase Inhibitor from Cajanus cajan Seeds: Purification, Characterization, and Insecticidal Properties

机译：Cajanus cajan种子的Bowman-Birk蛋白酶抑制剂：纯化，表征和杀虫特性。

获取原文
获取原文并翻译 | 示例
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

A red gram proteinase inhibitor (RgPI) was purified from red gram (Cajanus cajan) seeds by using ammonium sulfate precipitation and ion-exchange, affinity, and gel filtration chromatography. SDS-PAGE under nonreducing condition revealed two protein bands with molecular masses of ～8.5 and ～16.5 kDa corresponding to monomeric and dimeric forms of RgPI, respectively. Similarly, matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry also confirmed the presence of dimer as well as other oligomeric forms: trimer, tetramer, and pentamer. Reduction of RgPI with dithiothreitol (DTT) led to the dissociation of the dimeric and oligomeric forms. Native-PAGE and two-dimensional gel electrophoresis indicated the existence of isoinhibitors with p/ values of 5.95, 6.25, 6.50, 6.90, and 7.15, respectively. The MALDI-TOF-TOF mass spectrum and N-terminal sequence 'DQHHSSKACC suggested that the isolated RgPI is a member of the Bowman-Birk inhibitor family. RgPI exhibited noncompetitive type inhibitory activity against bovine pancreatic trypsin and chymo-trypsin, with inhibition constants of 292 and 2265 nM, respectively. It was stable up to a temperature of 80 °C and was active over a wide pH range between 2 and 12. However, reduction with DTT or 2-mercaptoethanol resulted in loss of inhibitory activity against trypsin and chymotrypsin. It also decreased the activity of larval midgut trypsin-like proteinases in Manduca sexta. Its insecticidal property was further confirmed by reduction in the growth and development of these larvae, when supplemented in the diet.
机译：通过使用硫酸铵沉淀和离子交换，亲和力和凝胶过滤色谱法，从红克（Cajanus cajan）种子中纯化了红克蛋白酶抑制剂（RgPI）。在非还原条件下的SDS-PAGE显示两个分子量分别为〜8.5和〜16.5 kDa的蛋白带，分别对应于RgPI的单体和二聚体形式。同样，基质辅助激光解吸电离飞行时间（MALDI-TOF）质谱法也证实了二聚体以及其他低聚形式：三聚体，四聚体和五聚体的存在。用二硫苏糖醇（DTT）还原RgPI导致二聚体和寡聚体形式解离。天然PAGE和二维凝胶电泳表明存在同种抑制剂，p /值分别为5.95、6.25、6.50、6.90和7.15。 MALDI-TOF-TOF质谱和N末端序列'DQHHSSKACC表明，分离的RgPI是Bowman-Birk抑制剂家族的成员。 RgPI对牛胰胰蛋白酶和胰凝乳蛋白酶表现出非竞争性抑制活性，抑制常数分别为292和2265 nM。它在高达80°C的温度下稳定，并且在2至12的宽pH范围内均具有活性。但是，用DTT或2-巯基乙醇还原会导致对胰蛋白酶和胰凝乳蛋白酶的抑制活性降低。它也降低了曼杜卡六倍体中幼虫中肠胰蛋白酶样蛋白酶的活性。当在日粮中添加这些幼虫时，其幼虫生长和发育的减少进一步证实了其杀虫性能。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号