Weiß Martin S, Pavlidis Ioannis V, Spurr Paul, Hanlon Steven P, Wirz Beat, Iding Hans, Bornscheuer Uwe T
Department of Biotechnology and Enzyme Catalysis, Institute of Biochemistry, University of Greifswald, Felix-Hausdorff-Strasse 4, 17489, Greifswald, Germany.
Group of Biotechnology, Department of Biochemistry, University of Kassel, Heinrich-Plett-Strasse 40, 34132, Kassel, Germany.
Chembiochem. 2017 Jun 1;18(11):1022-1026. doi: 10.1002/cbic.201700033. Epub 2017 Apr 27.
Amine transaminase (ATA) catalyzing stereoselective amination of prochiral ketones is an attractive alternative to transition metal catalysis. As wild-type ATAs do not accept sterically hindered ketones, efforts to widen the substrate scope to more challenging targets are of general interest. We recently designed ATAs to accept aromatic and thus planar bulky amines, with a sequence-based motif that supports the identification of novel enzymes. However, these variants were not active against 2,2-dimethyl-1-phenyl-propan-1-one, which carries a bulky tert-butyl substituent adjacent to the carbonyl function. Here, we report a solution for this type of substrate. The evolved ATAs perform asymmetric synthesis of the respective R amine with high conversions by using either alanine or isopropylamine as amine donor.
胺转氨酶（ATA）催化前手性酮的立体选择性胺化反应，是过渡金属催化的一种有吸引力的替代方法。由于野生型ATA不接受空间位阻较大的酮，因此将底物范围扩大到更具挑战性的目标的努力具有普遍意义。我们最近设计了ATA，使其能够接受芳香族以及平面型大体积胺，采用基于序列的基序来支持新型酶的鉴定。然而，这些变体对2,2-二甲基-1-苯基-丙-1-酮没有活性，该酮在羰基官能团附近带有一个庞大的叔丁基取代基。在此，我们报告了针对这类底物的解决方案。进化后的ATA通过使用丙氨酸或异丙胺作为胺供体，以高转化率进行相应R型胺的不对称合成。
