Yu Shuangrong, Qian Feng, Zhang Haimin, Sun Xinqiang, Wang Pu
Key Laboratory of Pharmaceutical Engineering of Zhejiang Province, College of Pharmaceutical Science, Zhejiang University of Technology, Hangzhou 310014, Zhejiang, China.
Key Laboratory for Green Pharmaceutical Technologies and Related Equipment of Ministry of Education, Zhejiang University of Technology, Hangzhou 310014, Zhejiang, China.
Sheng Wu Gong Cheng Xue Bao. 2024 Mar 25;40(3):821-833. doi: 10.13345/j.cjb.230478.
()-1-(2-fluorophenyl) ethylamine plays a crucial role as a chiral building block in pharmaceutical synthesis. ω-transaminases are widely recognized as environmentally friendly and efficient catalysts for the preparation of chiral amines. In this study, we isolated a novel ω-transaminase, TA, from through gene mining in the NCBI database. By employing semi-rational design, we obtained a Y168R/R416Q variant with enhanced enzyme activity. This variant exhibited the ability to catalyze the synthesis of ()-1-(2-fluorophenyl) ethylamine from 2-fluorophenone, achieving a yield of 83.58% and an enantioselectivity exceeding 99% after a 10 h reaction. Compared to the wild type, the specific enzyme activity of the Y168R/R416Q variant reached 47.04 U/mg, which represents an increase of 11.65 times. Additionally, the catalytic efficiency, as measured by /, was increased by 20.9 times. Molecular docking and structural simulation analysis revealed that the primary factor contributing to the improved catalytic efficiency is the expansion of the enzyme's active pocket and the alleviation of steric hindrance.
()-1-(2-氟苯基)乙胺作为手性结构单元在药物合成中起着关键作用。ω-转氨酶被广泛认为是制备手性胺的环境友好且高效的催化剂。在本研究中，我们通过在NCBI数据库中进行基因挖掘，从 中分离出一种新型的ω-转氨酶TA。通过采用半理性设计，我们获得了一种酶活性增强的Y168R/R416Q变体。该变体表现出能够催化从2-氟苯乙酮合成()-1-(2-氟苯基)乙胺的能力，在10小时反应后产率达到83.58%，对映选择性超过99%。与野生型相比，Y168R/R416Q变体的比酶活性达到47.04 U/mg，提高了11.65倍。此外，以/衡量的催化效率提高了20.9倍。分子对接和结构模拟分析表明，催化效率提高的主要因素是酶活性口袋的扩大和空间位阻的减轻。
