• 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Nanotechnology >Thermostable trypsin conjugates immobilized to biogenic magnetite show a high operational stability and remarkable reusability for protein digestion
【24h】

Thermostable trypsin conjugates immobilized to biogenic magnetite show a high operational stability and remarkable reusability for protein digestion

机译：固定在生物磁铁矿上的热稳定胰蛋白酶偶联物显示出高的操作稳定性和显着的蛋白质消化可重复使用性

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

In this work, magnetosomes produced by microorganisms were chosen as a suitable magnetic carrier for covalent immobilization of thermostable trypsin conjugates with an expected applicability for efficient and rapid digestion of proteins at elevated temperatures. First, a biogenic magnetite was isolated from Magnetospirillum gryphiswaldense and its free surface was coated with the natural polysaccharide chitosan containing free amino and hydroxy groups. Prior to covalent immobilization, bovine trypsin was modified by conjugating with α-, β- and γ-cyclodextrin. Modified trypsin was bound to the magnetic carriers via amino groups using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide and N-hydroxysulfosuccinimide as coupling reagents. The magnetic biomaterial was characterized by magnetometric analysis and electron microscopy. With regard to their biochemical properties, the immobilized trypsin conjugates showed an increased resistance to elevated temperatures, eliminated autolysis, had an unchanged pH optimum and a significant storage stability and reusability. Considering these parameters, the presented enzymatic system exhibits properties that are superior to those of trypsin forms obtained by other frequently used approaches. The proteolytic performance was demonstrated during in-solution digestion of model proteins (horseradish peroxidase, bovine serum albumin and hen egg white lysozyme) followed by mass spectrometry. It is shown that both magnetic immobilization and chemical modification enhance the characteristics of trypsin making it a promising tool for protein digestion.
机译：在这项工作中，由微生物产生的磁小体被选作合适的磁性载体，用于共价固定热稳定的胰蛋白酶结合物，并有望在高温下有效，快速地消化蛋白质。首先，从格氏螺螺旋藻中分离出一种生物磁铁矿，其自由表面涂有含有游离氨基和羟基的天然多糖壳聚糖。在共价固定之前，通过与α-，β-和γ-环糊精偶联来修饰牛胰蛋白酶。使用1-乙基-3-（3-二甲基氨基丙基）碳二亚胺和N-羟基磺基琥珀酰亚胺作为偶联剂，修饰的胰蛋白酶通过氨基与磁性载体结合。通过磁力分析和电子显微镜对磁性生物材料进行了表征。关于它们的生化特性，固定化的胰蛋白酶结合物显示出对高温的抵抗力增强，消除了自溶作用，具有最佳的pH值不变，并且具有明显的储存稳定性和可重复使用性。考虑到这些参数，提出的酶促系统显示出优于通过其他常用方法获得的胰蛋白酶形式的性质。蛋白溶解性能在模型蛋白（辣根过氧化物酶，牛血清白蛋白和鸡蛋清溶菌酶）的溶液中消化后进行了质谱分析。结果表明，磁性固定和化学修饰均增强了胰蛋白酶的特性，使其成为有希望的蛋白质消化工具。

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