基于计算的P450 MycG的机理研究和工程,用于16个成员的宏类抗生素的选择性氧化
Song Yang1, Matthew D DeMars2, Jessica M Grandner1
1Department of Chemistry and Biochemistry, University of California, Los Angeles, California 90095, United States.
Journal of the American Chemical Society
|October 8, 2020
概括
研究人员研究了MycG酶,一个P450细胞染色体,以了解其对化物M-IV的基质特异性. 计算和突变生成方法揭示了关键相互作用和改进的催化活性的工程变体.
科学领域:
- 生物化学
- 酵素学
- 计算化学
背景情况:
- MycG是一种细胞P450酶.
- 它催化了化物M-IV的两个连续氧化.
- 由于糖分的差异,MycG表现出M-IV比M-III和M-VI的优先氧化.
研究的目的:
- 研究MycG的化学选择性反应性和基质特异性.
- 了解MycG偏好M-IV的分子基础.
- 设计MycG变种,改变基质范围和产品配置.
主要方法:
- 密度函数理论 (DFT) 计算分析反应机制.
- 用于探索基质结合和特异性的元动力学和分子动力学 (MD) 模拟.
- 网站导向的突变发生,以验证计算预测和工程酶变体.
主要成果:
- DFT 发现基质环氧化因过度结合性损失而抑制C-H抽象.
- 在MD模拟中发现了对于基质识别至关重要的疏水糖结合口袋.
- 突变性实验证实了特定相互作用的作用,并设计了具有增强活性的MycG变体.
结论:
- 计算方法提供了MycG的催化机制和基质特异性的洞察力.
- 一个疏水口袋控制了基质的识别,这种特征在晶体结构中并不明显.
- 改造的MycG变种显示出对早期的mycinamicin前体的催化活性有所改善.
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