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.

创建具有催化生物转化能力的设计酶是一项艰巨的挑战。在最初的设计过程中，为实现这一目标所做的努力通常仅考虑规范氨基酸。但是，掺入具有独特反应性侧链特征的非天然氨基酸可能会大大扩展设计酶的催化范围。为了探索这种人工构建基团用于酶设计的潜力，在这里我们选择对氨基苯丙氨酸作为潜在的新型催化残基。我们证明that胺和侧链对和肟形成反应的催化活性是通过将对氨基苯丙氨酸嵌入多药转录调节剂的疏水孔中而增加的。乳酸乳球菌。通过混杂的结合口袋募集反应物和充当催化残基的明智放置的苯胺都有助于所鉴定的人造酶的成功。我们预计，我们的设计策略将在将来极大地扩展设计酶的催化范围方面带来回报。