Authors
Xiaole Yang, Xia Tian, Hai Zhu, Weiwei Jia, Jiahuan Li, Ruizhou Tang, Chuanzhi Zhang, Yang Cao, Tingting Li
Published in
Biotechnology letters. Volume 48. Issue 1. Pages 2. Nov 22, 2025. Epub Nov 22, 2025.
Abstract
Chiral amines, as essential chiral building blocks in drug synthesis, present a considerable challenge in biomanufacturing due to the requirement for highly efficient stereoselective synthesis. In this study, we successfully cloned and heterologously expressed a novel (R)-amine transaminase, MagAT, from Mycolicibacterium agri. Systematic analysis showed optimal activity at pH 7.0, with the highest reaction rate occurring within 30 min at 50 ℃. However, considering overall thermal stability, 40℃ was selected as the operating temperature for subsequent experiments. Furthermore, the enzyme retained nearly 100% catalytic activity in the presence of 10% methanol, DMSO, and chloroform. Kinetic analysis demonstrated that MagAT possessed high substrate affinity, with Michaelis constants (Km) of 5.13 ± 0.45 mM for (R)-1-phenylethan-1-amine and 3.73 ± 0.34 mM for pyruvate. Substrate specificity studies indicated that MagAT efficiently catalyzed the conversion of C3-C8 aliphatic amines (e.g., (R)-2-aminooctane) and monocyclic aromatic amines (e.g., (R)-1-phenylethan-1-amine), whereas its catalytic activity significantly declined for sterically hindered substrates such as naphthylamine. 3D structural modeling with AlphaFold3 and molecular docking showed that steric hindrance from key residues in the O-pocket critically influences substrate specificity. This study not only provides an efficient biocatalyst for the biosynthesis of chiral amines but also establishes a theoretical basis for the rational design of transaminases with broader substrate adaptability.
PMID:
41272324
Bibliographic data and abstract were imported from PubMed on 24 Nov 2025.
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