从前体前体的前体前体前体前体基因酸盐的酶合成
Wen-Shan Li1, Yingchun Li, Craig M Hill
1Department of Chemistry, Texas A&M University, P.O. Box 30012, College Station, Texas 77842-3012, USA.
Journal of the American Chemical Society
|April 4, 2002
概括
Pseudomonas diminuta 三酶可以选择性地切割特定的酶基质. 蛋白质工程允许相反的替代剂的水解,使得从前体前体的产物合成.
科学领域:
- 生物催化剂是一种生物催化剂.
- 酵素工程是什么? 酶工程是什么
- 有机合成 有机合成
背景情况:
- 三酶 (PTE) 酶对于排毒有机化合物至关重要.
- 来自Pseudomonas diminuta的酶在化基质中表现出酶选择性.
- 了解和操纵PTE的活性部位是控制其基质特异性的关键.
研究的目的:
- 为了研究 Pseudomonas diminuta phosphotriesterase (PTE) 对 bis-p-nitrophenyl 基酸盐的反选择性.
- 为了设计PTE变体,改变基质特异性,用于奇拉合成.
- 通过使用工程PTE来证明从前体前体中制备合产品的方法.
主要方法:
- 使用bis-p-nitrophenyl基酸酸的酶性水解试验.
- PTE活性部位的局部定向突变发生.
- 使用奇拉色谱分析产品的反体过量.
主要成果:
- 野生类型的PTE选择性地从各种基酸酸盐中切割了pro-R p-nitrophenolate替代物.
- 对于甲基,乙基和异基替代剂,产品的反体过量超过了99%.
- 产生了工程PTE变种,优先化了pro-S替代剂.
结论:
- Pseudomonas diminuta PTE 在特定基质水解方面表现出高的酶选择性.
- 在PTE活性位点的蛋白质工程允许反转enantioselectivity.
- 这种方法提供了一条新的途径,用于从亲基质中合成奇拉化合物的不对称合成.
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