利用色氨酸合酶的工程亚基合成β-支链色氨酸类似物
Synthesis of β-Branched Tryptophan Analogues Using an Engineered Subunit of Tryptophan Synthase.
作者信息
Herger Michael, van Roye Paul, Romney David K, Brinkmann-Chen Sabine, Buller Andrew R, Arnold Frances H
机构信息
Division of Chemistry and Chemical Engineering 210-41, California Institute of Technology , 1200 East California Boulevard, Pasadena, California 91125, United States.
出版信息
J Am Chem Soc. 2016 Jul 13;138(27):8388-91. doi: 10.1021/jacs.6b04836. Epub 2016 Jul 1.
Abstract
We report that l-threonine may substitute for l-serine in the β-substitution reaction of an engineered subunit of tryptophan synthase from Pyrococcus furiosus, yielding (2S,3S)-β-methyltryptophan (β-MeTrp) in a single step. The trace activity of the wild-type β-subunit on this substrate was enhanced more than 1000-fold by directed evolution. Structural and spectroscopic data indicate that this increase is correlated with stabilization of the electrophilic aminoacrylate intermediate. The engineered biocatalyst also reacts with a variety of indole analogues and thiophenol for diastereoselective C-C, C-N, and C-S bond-forming reactions. This new activity circumvents the 3-enzyme pathway that produces β-MeTrp in nature and offers a simple and expandable route to preparing derivatives of this valuable building block.
摘要
我们报道，在来自嗜热栖热菌的色氨酸合酶工程亚基的β-取代反应中，L-苏氨酸可以替代L-丝氨酸，一步生成(2S,3S)-β-甲基色氨酸(β-MeTrp)。通过定向进化，野生型β-亚基对该底物的微量活性提高了1000多倍。结构和光谱数据表明，这种增加与亲电氨基丙烯酸酯中间体的稳定性相关。该工程生物催化剂还能与多种吲哚类似物和苯硫酚发生反应，用于非对映选择性的C-C、C-N和C-S键形成反应。这种新活性绕过了自然界中产生β-MeTrp的三酶途径，为制备这种有价值的结构单元的衍生物提供了一条简单且可扩展的路线。
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