设计一种 (R) - 选择性转氨酶,用于 (R) - 3 - 氨基醇的不对称合成
He Liu1, Shixi Wang1, Meng Xu1
1State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, Shanghai 200237, China.
Bioorganic chemistry
|March 16, 2024
概括
工程化 (R) 选择性转氨酶显示了用于合成奇拉胺的增强的催化效率和稳定性. 这一突破改善了对具有挑战性的基质的酶接受,推动了药物发现.
科学领域:
- 生物催化和酶工程 生物催化和酶工程
- 奇拉胺胺的不对称合成
- 药物的发现和开发.
背景情况:
- 胺胺是小分子药物的关键组成部分.
- (R) 选择性转氨酶是有前途的催化剂,但在基质接受和效率方面存在局限性.
- 工程转氨酶是扩大它们在药物合成中的实用性的关键.
研究的目的:
- 来自Fodinicurvata sediminis的新型 (R) 选择性转氨酶 (FsTA) 的工程,以改善具有挑战性的基质的催化.
- 为了增强FsTA的基质结合,催化效率和热稳定性.
- 为创造具有更广泛基质范围的转氨酶变体提供合理的设计策略.
主要方法:
- 使用基底截断策略识别潜在的 (R) 选择性转氨酶 (FsTA).
- 计算分析 (分子对接和动力学) 以确定基质结合的关键残留物 (Y90).
- 代酶工程涉及活性部位重塑和共识序列策略,以生成改进的变体.
主要成果:
- 一个四倍突变 (H30R/Y90F/V152K/Y156F,mut4) 与初始变体相比,催化效率 (kcat/KM) 增加了7.95倍.
- 工程变体 (mut4) 的化温度 (Tm) 增加了10°C,这表明热稳定性得到了增强.
- Mut4对野生类型酶接受不良的各种子的活性显著提高.
结论:
- 理性酶工程可以克服 (R) 选择性转氨酶的局限性,提高它们的效率和基质范围.
- 开发的四重突变 (mut4) 提供了一个强大的生物催化剂,用于 valuable chiral amines 的不对称合成.
- 这项研究为设计能够在制药应用中容纳新和具有挑战性的基质的转氨酶奠定了基础.
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