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Structure‐ and Data‐Driven Protein Engineering of Transaminases for Improving Activity and Stereoselectivity

Ao, Yu‐Fei; Pei, Shuxin; Xiang, Chao; Menke, Marian J.; Shen, Lin; Sun, Chenghai; Dörr, Mark; Born, Stefan; Höhne, Matthias; Bornscheuer, Uwe T.

Amine transaminases (ATAs) are powerful biocatalysts for the stereoselective synthesis of chiral amines. Machine learning provides a promising approach for protein engineering, but activity prediction models for ATAs remain elusive due to the difficulty of obtaining high-quality training data. Thus, we first created variants of the ATA from Ruegeria sp. (3FCR) with improved catalytic activity (up to 2000-fold) as well as reversed stereoselectivity by a structure-dependent rational design and collected a high-quality dataset in this process. Subsequently, we designed a modified one-hot code to describe steric and electronic effects of substrates and residues within ATAs. Finally, we built a gradient boosting regression tree predictor for catalytic activity and stereoselectivity, and applied this for the data-driven design of optimized variants which then showed improved activity (up to 3-fold compared to the best variants previously identified). We also demonstrated that the model can predict the catalytic activity for ATA variants of another origin by retraining with a small set of additional data.
Published in: Angewandte Chemie International Edition, 10.1002/anie.202301660, Wiley-VCH