通过综合理性设计提高具有广泛基质范围的 (R) 选择性 ω-转氨酶的热稳定性
Peiyao Chang1,2, Xiao Li1,2, Zelin Lu2
1School of Biomedical Engineering (Suzhou), Division of Life Science and Medicine, University of Science and Technology of China, Hefei, Anhui 230026, China.
Journal of agricultural and food chemistry
|February 25, 2026
概括
研究人员设计了一种更稳定的 (R) 选择性欧米茄转氨酶 (R-ω-TA) 酶,用于奇拉胺合成. 这种改进的生物催化剂为工业应用提供了增强的热稳定性和催化效率.
科学领域:
- 生物催化和酶工程 生物催化和酶工程
- 有机合成 有机合成
- 蛋白质工程是指蛋白质工程.
背景情况:
- (R) 选择性欧米茄转氨酶 (R-ω-TAs) 对于合成奇拉氨酸至关重要,但它们的有限可用性和热不稳定性限制了工业用途.
- 天然R-ω-TAs的稀缺性和不良热稳定性对大规模的奇拉胺生产构成重大挑战.
研究的目的:
- 识别和设计一个热稳定的 (R) -选择性 ω-转氨酶,具有广泛的基质范围,用于工业生物催化.
- 为了提高 *Nocardioides* sp. 的新发现的 R-ω-TA 的热稳定性. 使用计算设计的CER19 (*Ns*TA).
主要方法:
- 来自 *Nocardioides* sp. 的一个新的 R-ω-TA (*Ns*TA) 的鉴定. CER19具有广泛的基板范围.
- 综合合理设计策略的应用,包括折叠自由能量预测,共识序列分析和表面电荷优化.
- 结构分析以阐明增强酶稳定性背后的机制.
主要成果:
- *Ns*TA的双重突变 (Y60F/V254L) 在45°C时半衰期增加了5倍,化温度 (Tm) 增加了7.82°C.
- 与野生类型酶相比,工程突变者保持了1.29倍更高的催化效率.
- 结构洞察力表明,改善的稳定性源于加强的界面疏水性和刚化的表面环.
结论:
- 这项研究成功地产生了一种高度热稳定的R-ω-TA突变体,具有广泛的基质范围,适合工业应用.
- 综合计算机辅助设计策略在设计强大的生物催化剂方面证明是有效的.
- 这项工作为通过合理设计开发稳定的工业酶提供了有价值的框架.
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