Hou Kaipeng, Huang Wei, Qi Miao, Tugwell Thomas H, Alturaifi Turki M, Chen Yuda, Zhang Xingjie, Lu Lei, Mann Samuel I, Liu Peng, Yang Yang, DeGrado William F
Department of Pharmaceutical Chemistry and Cardiovascular Research Institute, University of California, San Francisco, CA, USA.
Department of Chemistry and Biochemistry, University of California Santa Barbara, Santa Barbara, CA , USA.
Science. 2025 May 8;388(6747):665-670. doi: 10.1126/science.adt7268.
De novo design of protein catalysts with high efficiency and stereoselectivity provides an attractive approach toward the design of environmentally benign catalysts. Here, we design proteins that incorporate histidine-ligated synthetic porphyrin and heme ligands. Four of 10 designed proteins catalyzed cyclopropanation with an enantiomeric ratio greater than 99:1. A second class of proteins were designed to catalyze a silicon-hydrogen insertion and were optimized by directed evolution in whole cells. The evolved proteins incorporated features unlikely to be generated by computational design alone, including a proline in an α helix. Molecular dynamics simulations showed that as the proteins evolved toward higher activity, their conformational ensembles narrowed to favor more productive conformations. Our work demonstrates that efficient de novo protein catalysts are designable and should be useful for manifold chemical processes.
从头设计具有高效性和立体选择性的蛋白质催化剂为设计环境友好型催化剂提供了一种有吸引力的方法。在此，我们设计了包含组氨酸连接的合成卟啉和血红素配体的蛋白质。在设计的10种蛋白质中，有4种催化环丙烷化反应的对映体比例大于99:1。设计了第二类蛋白质来催化硅氢插入反应，并通过全细胞定向进化进行优化。进化后的蛋白质具有一些不太可能仅通过计算设计产生的特征，包括α螺旋中的一个脯氨酸。分子动力学模拟表明，随着蛋白质向更高活性进化，其构象集合变窄，有利于形成更具生产性的构象。我们的工作表明，高效的从头蛋白质催化剂是可以设计的，并且应该对多种化学过程有用。
