更快的表面结合反应改善了固定酶的结构和活性
Andres F Chaparro Sosa1, Riley M Bednar2, Ryan A Mehl2
1Department of Chemical and Biological Engineering, University of Colorado, Boulder, Colorado 80309, United States.
Journal of the American Chemical Society
|April 29, 2021
概括
在表面上的酶失活是个挑战. 更快的结合化学,如四素-sTCO,提高了酶固定效率,保护了酶结构和活性,以获得更好的生物混合材料.
科学领域:
- 生物混合材料科学
- 表面化学
- 酶固定化
背景情况:
- 材料表面的酶失活是生物混合材料的主要障碍.
- 表面变质热点会导致酶失活,限制材料的功能.
- 目前的策略侧重于酶稳定性,忽视了表面相互作用的动态.
研究的目的:
- 研究表面结合反应效率与酶结构/活性保留之间的联系.
- 确定更快的结合化学是否可以最大限度地减少化表面部位的酶探索.
- 提供设计更稳定和活跃的固定酶的见解.
主要方法:
- 使用快速四素 (Tet) 和紧张的转环 octene (sTCO) 结合化学.
- 使用动态单分子光追踪来监测酶表面相互作用.
- 将Tet-sTCO与较慢的化学物质 (thiol-maleimide,azide-dibenzocyclooctyne) 对酶固定进行比较.
主要成果:
- 碳酸无水酶 (CA) 与Tet- sTCO保持了77%的活性,相比之下,thiol- maleimide的活性为46%和azide- dibenzocyclooctyne的活性为27%.
- 较长的表面搜索距离 (> 0.5 μm) 显著增加了CA展开的概率.
- Tet-sTCO化学减少了CA固定距离,与更高的活动保留相关.
结论:
- 结合反应效率极大地影响了化表面热点的酶探索.
- 更快的结合化学减少了酶的展开,保持了结构和活性.
- 这种方法为创建强大的,功能性的生物混合材料提供了新的策略.
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