生物学杂志

• 技术方法 • 上一篇    下一篇

基于点击化学的蛋白酶修饰及其应用

  

  1. 江南大学 生态纺织教育部重点实验室, 无锡 214122
  • 出版日期:2019-06-18 发布日期:2019-06-18
  • 通讯作者: 王强,教授,主要从事纺织品生物加工技术研究,E-mail: qiang_wang@163.com
  • 作者简介:邵奕文,硕士研究生,主要研究方向为纺织生物技术,E-mail: shaoyiwen111@163.com
  • 基金资助:
    国家重点研发计划项目(2017YFB0309200);国家自然科学基金(51673087); 教育部长江学者和创新团队发展计划 (IRT_15R26)

Modification of protease via click chemistry and its application

  1. Key Laboratory of Eco-textiles, Ministry of Education, Jiangnan University, Wuxi 214122, China
  • Online:2019-06-18 Published:2019-06-18

摘要: 以葡聚糖为修饰剂,通过点击化学反应修饰蛋白酶(Savinase),增大蛋白酶分子量,从而更好地应用于羊毛生物法防毡缩加工。通过SDS-PAGE凝胶电泳、圆二色谱法和荧光光谱法考察蛋白酶修饰前后的结构变化,并测定其酶学性质和酶促动力学参数。结果表明,修饰后蛋白酶分子量明显增大,二级结构有所变化,酶与底物亲和力略有降低,但温度和pH值对酶活的影响情况相似。在羊毛防毡缩整理加工中,修饰蛋白酶处理的羊毛织物毡缩率满足“机可洗”标准,但强力损失明显减小。

关键词: 蛋白酶, 葡聚糖, 点击化学, 化学修饰, 羊毛防毡缩

Abstract: Savinase was modified with dextran via click chemistry so as to enlarge the molecular weight, which can be applied for shrink resistance of wool. The structure changes of modified Savinase were assessed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), circular dichroism (CD) and fluorescence spectroscopy. Enzymatic properties and kinetic parameters were also determined. The results indicated that the molecular weight of the modified Savinase was significantly increased. After modification, the secondary structure of enzyme changed and the substrate affinity lowered, while the effects of temperature and pH on enzyme activities were similar. For the shrink-resistance finishing of wool, it could be found that the wool fabric treated by modified Savinase could meet the machine-washable standard with obviously decreased strength loss.

Key words: protease, dextran, click chemistry, chemical modification, shrink resistance of wool

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