Ouyang Yao, Wang Suhao, Sorigue Damien, Hyster Todd K
Department of Chemistry, Princeton University, Princeton, New Jersey 08544, United States.
Aix-Marseille University, CEA, CNRS, Institute of Biosciences and Biotechnologies, BIAM Cadarache, 13108 Saint-Paul-lez-Durance, France.
J Am Chem Soc. 2025 Jul 23;147(29):25184-25190. doi: 10.1021/jacs.5c04097. Epub 2025 Jul 9.
Chiral amines are ubiquitous in pharmaceuticals and agrochemicals, making their efficient and selective synthesis a significant synthetic challenge. Threonine aldolases synthesize chiral amines via stereoselective C-C bond formation; however, they are restricted to small amino acids as pro-nucleophiles, limiting their utility in chemical synthesis. Here, we report an engineered threonine aldolase capable of α-functionalizing benzylamines. The evolved enzyme has excellent catalytic efficiency and accepts a broad range of (heterocyclic)benzyl amines and structurally diverse aldehydes to yield single-enantiomers of 1,2-amino alcohols in high-yield and diastereoselectivity. Mechanistic and crystallographic studies provide a rationale for how these mutations enable this previously unknown function. Moreover, beneficial mutations can be transferred to a related pyridoxal-dependent protein, highlighting the generality of these insights.
手性胺广泛存在于药物和农用化学品中，因此其高效且选择性的合成成为一项重大的合成挑战。苏氨酸醛缩酶通过立体选择性的碳-碳键形成来合成手性胺；然而，它们仅限于使用小氨基酸作为亲核试剂，这限制了它们在化学合成中的应用。在此，我们报道了一种能够对苄胺进行α-官能化的工程化苏氨酸醛缩酶。进化后的酶具有出色的催化效率，可接受多种（杂环）苄胺和结构多样的醛，以高产率和非对映选择性生成1,2-氨基醇的单一对映体。机理和晶体学研究为这些突变如何实现这种前所未有的功能提供了理论依据。此外，有益突变可以转移到相关的依赖于吡哆醛的蛋白质上，突出了这些见解的普遍性。
