SnoopLigase 催化-锁定并使固相结合体隔离成为可能
Can M Buldun1, Jisoo X Jean1, Michael R Bedford2
1Department of Biochemistry, University of Oxford , South Parks Road, Oxford, OX1 3QU, U.K.
Journal of the American Chemical Society
|February 7, 2018
概括
一种新型的蛋白质SnoopLigase通过同键有效连接标签 (SnoopTagJr和DogTag). 这项技术提高了酶的稳定性,并使生物结合和纳米组装的可重复使用催化剂成为可能.
科学领域:
- 蛋白质工程
- 生物结合化学
- 酶技术
背景情况:
- 连接生物分子的高效方法对于生物技术的发展至关重要.
- 蛋白质工程提供了一个强大的方法来创造具有特定功能的新型催化剂.
研究的目的:
- 设计和表征一种蛋白质连接酶 (SnoopLigase),以有效地形成特定位点的键.
- 设计与SnoopLigase一起使用的标签 (SnoopTagJr和DogTag).
- 证明SnoopLigase在增强酶稳定性和制造可重复使用的催化剂中的应用.
主要方法:
- 以结构,生物信息和计算分析为指导的蛋白质工程.
- 由SnoopLigase催化的地方特异性转amidation反应.
- 固相固定SnoopLigase用于催化剂重复使用.
- 酶循环增强耐热性
主要成果:
- 在SnoopTagJr和DogTag之间形成异键时,SnoopLigase的效率超过95%.
- SnoopTagJr和DogTag在包括内部部位在内的各种蛋白质位置上具有功能.
- 固相固定SnoopLigase允许高纯度产品的化和催化剂的再利用.
- 使用SnoopLigase的酶循环化为模型酶提供了特殊的耐热性 (高达100°C).
结论:
- SnoopLigase是一种高效的蛋白质结合和纳米组合工具.
- 开发的系统可以创建强大的可重复使用的生物催化剂.
- 这种方法提供了一种改善酶稳定性和功效的通用策略.
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