A designer enzyme for hydrazone and oxime formation featuring an unnatural catalytic aniline residue
A designer enzyme for hydrazone and oxime formation featuring an unnatural catalytic aniline residue
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DOI：
10.1038/s41557-018-0082-z
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发表时间：
2018-09-01
期刊：
影响因子：
21.8
通讯作者：
Roelfes, Gerard
中科院分区：
文献类型：
--
作者：
Drienovska, Ivana;Mayer, Clemens;Roelfes, Gerard
创建具有催化物质转化能力的设计师酶是一个巨大的挑战。实现此目标的努力通常仅考虑最初的设计过程中的规范氨基酸。但是，结合具有独特反应性侧链的非天然氨基酸可以显着扩大设计酶的催化库。为了探索这种人工构建块在酶设计中的潜力，在这里，我们选择了P-氨基苯胺作为潜在的新型催化残基。我们证明，通过将p-氨基苯甲酰氨酸嵌入来自乳酸乳酸乳球菌的多剂量转录调节剂的疏水孔中，通过将P-氨基苯胺的苯胺侧链和氧赛形成反应的催化活性增加。通过混合袋对反应物的募集和明智地放置的苯胺，作为催化残基的作用都有助于确定的人工酶的成功。我们预计，我们的设计策略将获得回报，以显着扩大未来设计师酶的催化曲目。
Creating designer enzymes with the ability to catalyse abiological transformations is a formidable challenge. Efforts toward this goal typically consider only canonical amino acids in the initial design process. However, incorporating unnatural amino acids that feature uniquely reactive side chains could significantly expand the catalytic repertoire of designer enzymes. To explore the potential of such artificial building blocks for enzyme design, here we selected p-aminophenylalanine as a potentially novel catalytic residue. We demonstrate that the catalytic activity of the aniline side chain for hydrazone and oxime formation reactions is increased by embedding p-aminophenylalanine into the hydrophobic pore of the multidrug transcriptional regulator from Lactococcus lactis. Both the recruitment of reactants by the promiscuous binding pocket and a judiciously placed aniline that functions as a catalytic residue contribute to the success of the identified artificial enzyme. We anticipate that our design strategy will prove rewarding to significantly expand the catalytic repertoire of designer enzymes in the future.