In this study, we present a significant advancement in the field of enzymatic asymmetric reductive amination (ARA) of ketones, a pivotal reaction for chiral amine synthesis. Through a combination of semirational enzyme design and bioprocess development, we achieve the dual activation and stabilization of amine dehydrogenase (AmDH) to meet industrial demands. The engineered AmDH exhibits remarkable catalytic efficiency (turnover number, TON >1,000,000) and exceptional stability (half-life >7 days at 50 °C), with a broadened substrate scope including various aryl alkyl ketones and fatty ketones. Leveraging biobased oleic acid as an activator and stabilizer, we achieved kilogram-scale synthesis of chiral amines. Furthermore, the integration of AmDH with chemical catalysts in chemoenzymatic cascades has enabled the synthesis of a wide array of pharmaceutically relevant amines from diverse substrates, demonstrating the enzyme’s versatility and potential to transform synthetic chemistry.

中文翻译：

---

脂肪酸活化和稳定工程胺脱氢酶用于不对称还原胺化的生物过程强化


在这项研究中，我们提出了酮的酶促不对称还原胺化 （ARA） 领域的重大进展，这是手性胺合成的关键反应。通过半合理的酶设计和生物工艺开发的结合，我们实现了胺脱氢酶 （AmDH） 的双重活化和稳定化，以满足工业需求。工程化的 AmDH 表现出卓越的催化效率（周转数，吨 >1,000,000）和出色的稳定性（50 °C 下半衰期 >7 天），并扩大了底物范围，包括各种芳基烷基酮和脂肪酮。利用生物基油酸作为活化剂和稳定剂，我们实现了手性胺的公斤级合成。此外，AmDH 与化学酶级联反应中的化学催化剂的整合使得从不同底物合成多种药学相关胺成为可能，证明了该酶的多功能性和改变合成化学的潜力。