使用ProteinMPNN指导工程开发一种高功能的热稳定的D-氨基酸氧化酶,用于高效的L-phosphinothricin生物合成
Kai Yang1, Yueshan Huang1, Liyi Yao1
1Central South University, School of Minerals Processing and Bioengineering, 932 South Lushan Road, Yuelu District, Changsha, Hunan, 410083, PR China.
International journal of biological macromolecules
|December 19, 2025
概括
改造的D-氨基酸氧化酶 (DAAO) 显示了对D-葡萄糖酸盐转换的增强稳定性和效率. 这种蛋白质工程策略推进了性氨基酸生物合成,并降低了查成本.
科学领域:
- 生物催化剂是一种生物催化剂.
- 蛋白质工程是指蛋白质工程.
- 酵素进化 酵素进化
背景情况:
- 由于其立体特异性,D-氨基酸氧化酶 (DAAO) 是性氨基酸合成的关键酶.
- DAAO的有限热稳定性阻碍了其工业应用和蛋白质工程的努力.
研究的目的:
- 为了设计一个DAAO变体,提高催化效率和热稳定性.
- 扩大DAAO的基质范围,用于非天然氨基酸合成.
主要方法:
- 以蛋白MPNN为导向的多重密码子和突变性被用来设计DAAO.
- 工程变体的催化效率,热稳定性和基质范围的表征.
- 用分子动力学 (MD) 模拟来理解增强性能的结构基础.
主要成果:
- 改造的DAAO变体 (M5) 显示了对D-glufosinate (D-PPT) 的催化效率增加了29倍.
- 这种变种在热稳定性方面呈现了208倍的增强,其半衰期在50°C时为519分钟.
- 设计的DAAO在7个小时内成功地在100毫升尺度上以高立体特异性转换了500毫米的D-PPT.
结论:
- 工程设计的DAAO变种在催化效率和热稳定性方面取得了显著的改进,使其适合工业应用.
- 该研究提供了对提高生物催化剂性能的酶工程策略的见解.
- 这种方法提供了一种具有成本效益的方法,用于用于有价值产品生物合成的酶的定向进化.
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