通过半理性设计,增强BpIH的伊马达酶活性,使其向3-异布基利化物转移
Wenping Qin1,2, Long Xu3, Kun Cheng1
1School of Pharmaceutical Sciences, Taizhou University, 1139 Shifu Rd, Taizhou, 318000, China.
Applied microbiology and biotechnology
|September 25, 2024
概括
一种新的酶突变,YHS-I,显著增强了 (R) -3-异布基酸单胺 (R-IBM) 的产生,这是一种用于普雷加巴林合成的关键中间体. 这种工程造影酶实现了高的转换率和反体过量,改善了工业制药的制造.
科学领域:
- 生物催化和酶工程 生物催化和酶工程
- 有机合成 有机合成
- 制药化学 制药化学 制药化学
背景情况:
- 来自Burkholderia phytofirmans的酶BpIH可以有效地产生 (R) -3-异布基酸单胺 (R-IBM).
- R-IBM是合成止痛药pregabalin的一个关键中间体.
- 野生型BpIH从3-异布基利胺 (IBI) 获得了100%的理论产量.
研究的目的:
- 使用同质模型阐明BpIH的基质结合口袋.
- 设计BpIH以提高R-IBM生产中的催化活性和效率.
- 为了研究改善酶性能的结构基础.
主要方法:
- 同一性建模和结构对齐以识别基质结合口袋.
- 基于氨基酸残留保存分析的半理性酶设计.
- 现场定向的突变发生,分子动力学和对接模拟.
主要成果:
- 一种三重突变,Y37FH133NS226I (YHS-I),表明酶活性增加了五倍.
- YHS-I实现了IBI到R-IBM的88.87%的转换率,其反体过量>99.9%.
- 突变增强了酶刚性和基质亲和力,从而提高了催化效率 (kcat/Km).
结论:
- 半理性设计的BpIH可以显著提高其催化性能.
- 这种YHS-I突变体代表了用于R-IBM生产的高效生物催化剂.
- 工程酶为优化制药中间体合成提供了一个有前途的途径.
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