7YPM

Crystal structure of transaminase CC1012 complexed with PLP and L-alanine

7YPM の概要

• エントリーDOI: 10.2210/pdb7ypm/pdb
• 分子名称: Aspartate aminotransferase family protein, PYRIDOXAL-5'-PHOSPHATE, ALANINE, ... (6 entities in total)
• 機能のキーワード: complex, tranaminase, transferase
• 由来する生物種: Caulobacter sp. D5
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 209464.33

構造登録者

Yang, L.,Wang, H.,Wei, D. (登録日: 2022-08-03, 公開日: 2023-05-24, 最終更新日: 2023-11-29)

主引用文献

Yang, L.,Zhang, K.,Xu, M.,Xie, Y.,Meng, X.,Wang, H.,Wei, D.
Mechanism-Guided Computational Design of omega-Transaminase by Reprograming of High-Energy-Barrier Steps.
Angew.Chem.Int.Ed.Engl., 61:e202212555-e202212555, 2022

PubMed Abstract: ω-Transaminases (ω-TAs) show considerable potential for the synthesis of chiral amines. However, their low catalytic efficiency towards bulky substrates limits their application, and complicated catalytic mechanisms prevent precise enzyme design. Herein, we address this challenge using a mechanism-guided computational enzyme design strategy by reprograming the transition and ground states in key reaction steps. The common features among the three high-energy-barrier steps responsible for the low catalytic efficiency were revealed using quantum mechanics (QM). Five key residues were simultaneously tailored to stabilize the rate-limiting transition state with the aid of the Rosetta design. The 14 top-ranked variants showed 16.9-143-fold improved catalytic activity. The catalytic efficiency of the best variant, M9 (Q25F/M60W/W64F/I266A), was significantly increased, with a 1660-fold increase in k /K and a 1.5-26.8-fold increase in turnover number (TON) towards various indanone derivatives.

PubMed: 36300723
DOI: 10.1002/anie.202212555

実験手法

X-RAY DIFFRACTION (1.984 Å)