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In parallel, advances in deep-learning-driven\n                  <jats:italic>de novo<\/jats:italic>\n                  protein design have expanded accessible protein architectural space, enabling the generation of functional catalysts for increasingly challenging transformations. We herein report a synergistic design-evolution strategy to encode new-to-nature regio- and enantioselective C\u2013H amination reactions using engineered\n                  <jats:italic>de novo<\/jats:italic>\n                  haemproteins to synthesise chiral piperidines, a privileged pharmacophore in drug discovery. By integrating motif scaffolding, computational re-design, and directed evolution, we demonstrate\n                  <jats:italic>de novo<\/jats:italic>\n                  enzyme design can enable challenging, high-energy-barrier enzyme chemistry with programmable regio- and enantioselectivity, high catalytic efficiency, excellent enzyme stability, and scaffold diversity.\n                <\/jats:p>","DOI":"10.64898\/2026.10.01.754751","type":"posted-content","created":{"date-parts":[[2026,10,2]],"date-time":"2026-10-02T21:10:23Z","timestamp":1790975423000},"source":"Crossref","is-referenced-by-count":0,"title":["Design-Evolution Synergy Enables New-to-Nature Enzyme Chemistry"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-1353-3004","authenticated-orcid":false,"given":"Zi-Qi","family":"Li","sequence":"first","affiliation":[{"name":"Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena, California 91125, United 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Manuscript in preparation. In this work, we demonstrate the complementary regioselectivity for the synthesis of chiral pyrrolidines through directed evolution of a natural haemprotein."},{"key":"2026100620254845000_2026.10.01.754751v1.68","doi-asserted-by":"publisher","DOI":"10.1021\/jacs.3c08053"},{"key":"2026100620254845000_2026.10.01.754751v1.69","doi-asserted-by":"publisher","DOI":"10.1038\/s41589-026-02235-9"},{"key":"2026100620254845000_2026.10.01.754751v1.70","doi-asserted-by":"publisher","DOI":"10.1021\/jacs.2c08285"},{"key":"2026100620254845000_2026.10.01.754751v1.71","doi-asserted-by":"publisher","DOI":"10.1126\/science.abk1603"},{"key":"2026100620254845000_2026.10.01.754751v1.72","doi-asserted-by":"publisher","DOI":"10.1016\/j.bpj.2014.05.009"}],"container-title":[],"original-title":[],"link":[{"URL":"https:\/\/syndication.highwire.org\/content\/doi\/10.64898\/2026.10.01.754751","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,10,7]],"date-time":"2026-10-07T03:26:03Z","timestamp":1791343563000},"score":1,"resource":{"primary":{"URL":"http:\/\/biorxiv.org\/lookup\/doi\/10.64898\/2026.10.01.754751"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,10,2]]},"references-count":72,"URL":"https:\/\/doi.org\/10.64898\/2026.10.01.754751","relation":{},"subject":[],"published":{"date-parts":[[2026,10,2]]},"subtype":"preprint"}}