基于多维理性设计的 Echinocandin B 脱酶的疏水基质结合口袋改造
Heng Tang1, Han-Yue Zhu1, Yin-Feng Huang1
1National and Local Joint Engineering Research Center for Biomanufacturing of Chiral Chemicals, Zhejiang University of Technology, Hangzhou 310014, PR China; Key Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou 310014, PR China.
International journal of biological macromolecules
|April 13, 2024
概括
酶工程显著提高了Actinoplanes utahensis deacylase (AAC) 对抗真菌合成的活性. 量身定制基质结合口袋提高了催化效率,使得安尼杜拉金生产的产品转化率更高.
科学领域:
- 生物催化剂是一种生物催化剂.
- 酶工程是什么? 酶工程是什么?
- 药用化学 医学化学
背景情况:
- 安尼杜拉金是一种赤甘丁抗真菌剂,通过使用Actinoplanes utahensis deacylase (AAC) 脱氧化赤甘丁B (ECB) 来合成.
- 野生类型AAC的低催化活性限制了其在anidulafungin合成中的工业应用.
研究的目的:
- 提高AAC的催化效率,以改善无类菌素的生产.
- 通过合理设计设计AAC,以更好地适应复杂的基板ECB.
主要方法:
- 多维理性设计策略侧重于基板结合口袋体积,曲率和长度.
- 基质性质的定量分析 (水友性和疏水性).
- 在AAC.的酸结合口袋中的关键残留物的位点定向突变发生.
主要成果:
- 工程 AAC 突变物成功地适应了 ECB 的延长脂质链,具有较低的曲率结合口袋.
- 与野生型AAC相比,D53A/I55F/G57M/F154L/Q661L突变 (MT) 的催化效率增加了331%.
- MT实现了94.6%的产品转化率,这是该酶反应报告的最高水平.
结论:
- 多维理性设计是一种有效的策略,用于提高复杂基质的酶催化效率.
- 调整AAC的基质结合口袋显著提高了其在anidulafungin合成中的性能.
- 开发的AAC突变提供了一个有前途的生物催化解决方案,用于高效的anidulafungin生产.
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