对CYP450融合蛋白 (CYPLY) 的基质耐受机制和域协同工程的动态分析
Yisang Zhang1,2, Yuanwei Wang1,2, Huiying Zhu1,2
1State Key Laboratory of Green Papermaking and Resource Recycling, Qilu University of Technology, Jinan 250353, China.
Journal of agricultural and food chemistry
|December 22, 2025
概括
工业用途的细胞染色体P450 (CYP450) 酶工程通过了解基质抑制和电子转移而得到了推进. 这项研究揭示了一种协同模型,将关门动态与提高酶性能联系起来,并确定了用于增强催化力的关键突变.
科学领域:
- 生物催化和酶工程 生物催化和酶工程
- 计算生化学计算生化学
- 工业生物技术 工业生物技术
背景情况:
- 细胞染色体P450 (CYP450) 酶对于天然产品生物合成和工业催化是至关重要的.
- 工程 CYP450s 的关键挑战包括克服基质抑制和提高电子转移效率.
- 了解酶结构,动力学和功能之间的相互作用对于有针对性的工程至关重要.
研究的目的:
- 阐明化学CYP450酶中基质道的封闭机制及其与基质抑制的联系.
- 研究构造动态在CYP450系统中调节电子转移效率中的作用.
- 为工业应用设计一种具有减轻基质抑制和改善催化性能的仿真CYP450酶.
主要方法:
- 集成的多模板同质模型和分子动力学 (MD) 模拟来分析酶结构和动力学.
- 使用基于深度学习的突变预测来识别有益的氨基酸替代物.
- 用于基质抑制,电子转移效率和整体催化转换的特征酶变体.
主要成果:
- 阐明了基质道封闭机制与基质抑制之间的直接联系,在仿真CYP153A/M228L-CPRBM3 (CYPLY) 酶中.
- 鉴定了CYPLY-N243D变异,该变异减少了基质抑制的34倍.
- 开发了一种双重突变的CYPLY-N243D/T592N,表现出基质抑制减少了61倍,转化率为65.8%,反应时间减少了4小时.
结论:
- 为工业CYP450工程建立了一种涉及门,动力学和电子转移的协同监管模型.
- 证明了结合计算建模和深度学习的有效性,以实现合理的酶设计.
- 工程化CYP450变体显示出改善工业中的生物催化工艺的巨大潜力.
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