纤维素脱酶的接口工程改善了域间电子转移
Thomas M B Reichhart1,2, Stefan Scheiblbrandner1, Christoph Sygmund2
1Biocatalysis and Biosensing Laboratory, Institute of Food Technology, Department of Food Science and Technology, University of Natural Resources and Life Sciences (BOKU), Vienna, Austria.
Protein science : a publication of the Protein Society
|June 14, 2023
概括
接口工程通过在生理pH下增强域间电子转移 (IET) 来改善生物传感器的细胞质脱酶 (CDH). 然而,修改减少了直接电子转移 (DET),需要进一步优化生物电子应用.
科学领域:
- 生物催化剂是一种生物催化剂.
- 生物电化学 生物电化学
- 蛋白质工程是指蛋白质的工程.
背景情况:
- 蜂脱酶 (CDH) 是生物传感器和生物燃料电池中直接电子转移 (DET) 的关键生物电触媒.
- 它在生理pH值下具有最佳的酸性pH值和缓慢的域间电子转移 (IET),限制了其在葡萄糖测量中的应用.
- 脱酶和细胞染色体域 (CYT) 之间的界面上的静电排斥阻碍了电子传输.
研究的目的:
- 设计CDH酶接口以在生理pH下加速IET.
- 研究改进IET背后的机制,并确定未来生物电子应用的策略.
主要方法:
- 通过突变酸性氨基酸来进行CYT领域的理性接口工程.
- 植物遗传学和结构分析以指导变体设计.
- 17种CDH变体的pH最佳,IET速率和DET的表征.
主要成果:
- 五个单个突变 (G71K,D160K,Q174K,D177K,M180K) 提高了pH最佳值和IET速率.
- 组合变体将pH最佳值转移到7.0并且在pH7.5时增加IET超过12倍.
- 在CYT上增加的积极费用改善了IET,但减少了DET,表明了权衡.
结论:
- 接口工程是有效的优化CDH的pH最佳和IET.
- 静电转向和键稳定了酶的封闭状态,增强了IET.
- 保持DET对于工程CDH的未来生物电子应用至关重要.
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