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Solid-phase Synthesis of [4.4] Spirocyclic Oximes
Published on: February 6, 2019
循环氨单氧化酶的结构导向进化向大容量欧米硫化物:基质迁移和立体选择性反转
Shiyu Wei1,2, Guochao Xu2, Jieyu Zhou2
1Institution: School of Biological and Pharmaceutical Engineering, Lanzhou Jiaotong University, Address, Lanzhou Jiaotong University, Lanzhou, 730070, Gansu, China.
结构引导的工程 Amycolatopsis methanolica CHMO (AmCHMO) 增强的不对称硫化. 突变将立体选择性从R逆转为S,为梅硫化物生产实现了高反体过量.
科学领域:
- 生物催化和酶工程 生物催化和酶工程
- 有机化学 有机化学
- 分子生物学分子生物学
背景情况:
- 奇拉药物合成依赖于对抗选择性催化剂.
- 循环松单氧酶 (CHMOs) 是多功能生物催化剂.
- 针对特定基质转换的工程CHMO具有挑战性.
研究的目的:
- 为了改进Amycolatopsis methanolica CHMO (AmCHMO) 来改善硫化的不对称硫化.
- 通过结构导向突变实现高立体选择性和活性.
- 调查改变基质特异性和立体选择性的结构基础.
主要方法:
- 组合活性位点和试验 (CASTing) 和代和变异 (ISM).
- 结构引导工程专注于基板道和结合口袋残留物.
- 分子动力学 (MD) 模拟来分析反应前状态.
主要成果:
- 工程突变MT3显示高R-立体选择性 (99%) 和活性 (46.19U/g).
- 进一步的突变产生了MT8与倒置的S-立体选择性 (97%),但减少了活动.
- 突变MT11结合了有益的突变,实现了2.29 U/g的活性和97%的S-立体选择性.
- MD模拟与观察到的活动和立体选择性相关的结构变化.
结论:
- 扩大基质道扩大了基质光谱,而重塑结合口袋则逆转了立体选择性.
- 结构引导工程可以精确控制CHMO活动和立体选择性.
- 这项工作为工程学Baeyer-Villiger单氧化酶 (BVMOs) 提供了一个框架,用于大容量乙烯氧化.
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