氧化还原可逆人造胺基还原酶的动态和结构分析
Alex H Miller1, Ingrid B S Martins2, Elena V Blagova3
1Department of Chemistry, University of York, Heslington, York YO10 5DD, United Kingdom.
三种人造胺基还原酶 (ArM) 显示出不同的催化效率. 动力学和对接研究揭示了基质特异性相互作用和多个结合位点,解释了性能和中度的酶选择性差异.
科学领域:
- 生物催化和酶工程 生物催化和酶工程
- 有机金属化学 有机金属化学
- 超分子化学 超分子化学
背景情况:
- 人工金属酶 (ArM) 是一种工程催化剂,它将蛋白质支架与金属辅助因子结合起来.
- 使用Fe(III) - 亚托切林的超分子定已被用于将催化剂固定在周等离子体结合蛋白 (PBP) 支架上.
- 之前的研究合成了三种同类的ARM,但观察到它们的催化周转频率 (TOF) 的显著变化.
研究的目的:
- 为了研究三种人工 imine 减少酶 (ArM) 与不同循环 imine 基质的动态行为.
- 阐明了导致同类ARM之间观察到的催化效率变化的因素.
- 了解基质结合相互作用及其对催化活性和酶选择性的影响.
主要方法:
- 使用三种循环 imine (二化素,哈马林,帕帕维林) 来确定迈凯利斯-门参数的ARM的动态分析.
- 分子对接模拟用于预测人工结合口袋内的基质蛋白相互作用.
- 评估未经修改的PBP支架所赋予的enantioselectivity. 没有修改的PBP支架.
主要成果:
- 这三种ARM都遵循迈凯利斯-门动力学,哈马林表现出最明显的和度.
- 分子对接揭示了基质和ArM残留物之间的多种结合,pi堆积和pi离子相互作用.
- 基质哈马林和帕帕维林的亲和力比二化素更高,与对接预测一致.
- 在催化中心附近确定了多个基质容纳腔.
结论:
- 人工伊米因还原酶的动态行为和结合相互作用是基质依赖的.
- 同类ARM之间的TOFs的变化可以归因于基质结合亲和力和方向的差异.
- 多个结合口袋的存在影响了这些ARM的催化效率和中等的反选择性.
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