De novo design of porphyrin-containing proteins as efficient and stereoselective catalysts

• 成果类型: Article
• 署名作者: 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.
• 署名单位: University of California System; University of California San Francisco; University of California System; University of California San Francisco; University of California System; University of California Santa Barbara; Pennsylvania Commonwealth System of Higher Education (PCSHE); University of Pittsburgh; University of California System; University of California Riverside; University of California System; University of California Santa Barbara
• 刊物名称: SCIENCE
• ISSN/ISSBN: 0036-12997
• DOI: 10.1126/science.adt7268
• 发表日期: 2025-05-08
• 页码: 665-670
• 关键词: molecular-dynamics; carbene transfer; computational design; directed evolution; electron-transfer; atomic charges; heme-proteins; cytochrome-c; enzymes; energy

摘要

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 alpha 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.