Computational-Guided Hinge-Region Engineering Unlocks QtPheDH for Synthesizing Bulky Naphthyl Amino Acids
ACS Sustainable Chemistry & Engineering, vol. 13, pp. 12542–12552
Abstract
Asymmetric reductive amination of α-keto acids catalyzed by amino acid dehydrogenases (AADHs) provides an atom-efficient and sustainable strategy for synthesizing optically pure non-natural amino acids. However, the narrow substrate spectrum of AADHs often restricts their broader application in green synthesis. Herein, a functional hinge region composed of three helices was identified as a key structural determinant governing substrate recognition and entry in both phenylalanine and leucine dehydrogenases. By engineering the hinge region, the catalytic performance of Qt PheDH, a phenylalanine dehydrogenase from Quasibacillus thermotolerans, was significantly improved. Specifically, variant M2(V311G/S158G) alleviated the steric barrier and facilitated the entry of bulky substrates, enabling the initial amination activity of 1-naphthylglyoxylic acid ( 5a ). Further optimization through binding energy reconfiguration led to the development of M4.2(V311G/S158G/L308M/T126I), which exhibited a 10-fold increase in catalytic efficiency and achieved complete conversion of 100 mM 5a with >99% ee . MD simulations revealed that the enhanced conformational flexibility in the active pocket and widened substrate tunnel were indispensable for accommodating bulkier substrates. These findings highlight the potential of hinge-region engineering in evolving AADHs toward powerful biocatalysts for sustainable and scalable synthesis of bulky non-natural amino acids.
Authors 7
-
Affiliation as printed
Jiangnan University
Key laboratory of industrial Biotechnology, Ministry of Education, School of Biotechnology
State Key Laboratory of Food Science and Resources
State Key Laboratory of Food Science and Resources, Jiangnan University, Wuxi 214122, Jiangsu, China
-
Affiliation as printed
Jiangnan University
Key laboratory of industrial Biotechnology, Ministry of Education, School of Biotechnology
State Key Laboratory of Food Science and Resources
State Key Laboratory of Food Science and Resources, Jiangnan University, Wuxi 214122, Jiangsu, China
-
Affiliation as printed
Jiangnan University
Key laboratory of industrial Biotechnology, Ministry of Education, School of Biotechnology
State Key Laboratory of Food Science and Resources
State Key Laboratory of Food Science and Resources, Jiangnan University, Wuxi 214122, Jiangsu, China
-
Ulrich Schwaneberg Aachen DWI - Leibniz Institute for Interactive Materials Institute of Biotechnology
DWI – Leibniz Institute for Interactive Materials · RWTH Aachen University
Affiliation as printed
DWI-Leibniz Institute for Interactive Materials
Institute of Biotechnology
RWTH Aachen University
Institute of Biotechnology, RWTH Aachen University, Aachen 52074, Germany
-
Affiliation as printed
Jiangnan University
Key laboratory of industrial Biotechnology, Ministry of Education, School of Biotechnology
State Key Laboratory of Food Science and Resources
State Key Laboratory of Food Science and Resources, Jiangnan University, Wuxi 214122, Jiangsu, China
-
Guochao Xu corresponding
Jiangnan University · Shanghai University of Traditional Chinese Medicine
Affiliation as printed
Jiangnan University
Key laboratory of industrial Biotechnology, Ministry of Education, School of Biotechnology
Shanghai University of Traditional Chinese Medicine
The Research Center of Chiral Drugs, Shanghai Frontiers Science Center for TCM Chemical Biology, Innovation Research Institute of Traditional Chinese Medicine
Key laboratory of industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, Jiangsu, China
-
Ye Ni corresponding
Affiliation as printed
Jiangnan University
Key laboratory of industrial Biotechnology, Ministry of Education, School of Biotechnology
State Key Laboratory of Food Science and Resources
State Key Laboratory of Food Science and Resources, Jiangnan University, Wuxi 214122, Jiangsu, China
Cited by 1 stored of 1
1 result
No patents citing this paper on Lens.org (checked 2026-10-06).