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A succession of TrpB protein engineering campaigns to expand the enzyme's nucleophile substrate range has enabled the biocatalytic production of diverse non\u2010canonical amino acids (ncAAs). Here, we show that ketone\u2010derived enolates can serve as nucleophiles in the TrpB reaction to achieve the asymmetric alkylation of ketones, an outstanding challenge in synthetic chemistry. We engineered TrpB by directed evolution to catalyze the asymmetric alkylation of propiophenone and 2\u2010fluoroacetophenone with a high degree of selectivity. In reactions with propiophenone, preference for the opposite product diastereomer emerges over the course of evolution, demonstrating that full control over the stereochemistry at the new chiral center can be achieved. The addition of this new reaction to the TrpB platform is a crucial first step toward the development of efficient methods to synthesize non\u2010canonical prolines and other chirally dense nitrogen heterocycles.<\/jats:p>","DOI":"10.1002\/anie.202106938","type":"journal-article","created":{"date-parts":[[2021,7,16]],"date-time":"2021-07-16T02:09:41Z","timestamp":1626401381000},"page":"21412-21417","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":30,"title":["Asymmetric Alkylation of Ketones Catalyzed by Engineered TrpB"],"prefix":"10.1002","volume":"60","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-0585-1598","authenticated-orcid":false,"given":"Ella J.","family":"Watkins\u2010Dulaney","sequence":"first","affiliation":[{"name":"Division of Biology and Biological Engineering California Institute of Technology, MC 210-41  1200 E. 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