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While these enzymes offer an appealing alternative to traditional Baeyer\u2010Villiger oxidations, these proteins tend to be either too unstable or exhibit too narrow of a substrate scope for implementation as industrial biocatalysts. Here, sequence similarity networks were used to search for novel BVMOs that are both stable and promiscuous. Our genome mining led to the identification of an enzyme from Chloroflexota bacterium (strain G233) dubbed\n                    <jats:italic>ssn<\/jats:italic>\n                    BVMO that exhibits i) the highest melting temperature of any naturally sourced BVMO (62.5\u2009\u00b0C), ii) a remarkable kinetic stability across a wide range of conditions, similar to those of PAMO and PockeMO, iii) optimal catalysis at 50\u2009\u00b0C, and iv) a broad substrate scope that includes linear aliphatic, aromatic, and sterically bulky ketones. Subsequent quantitative assays using propiophenone demonstrated &gt;95\u2009% conversion. Several fusions were also constructed that linked\n                    <jats:italic>ssn<\/jats:italic>\n                    BVMO to a thermostable phosphite dehydrogenase. These fusions can recycle NADPH and catalyze oxidations with sub\u2010stoichiometric quantities of this expensive cofactor. Characterization of these fusions permitted identification of PTDH\u2212L1\u2010\n                    <jats:italic>ssn<\/jats:italic>\n                    BVMO as the most promising protein that could have utility as a seed sequence for enzyme engineering campaigns aiming to develop biocatalysts for Baeyer\u2010Villiger oxidations.\n                  <\/jats:p>","DOI":"10.1002\/cbic.202400443","type":"journal-article","created":{"date-parts":[[2024,7,11]],"date-time":"2024-07-11T16:25:41Z","timestamp":1720715141000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Genome Mining Leads to the Identification of a Stable and Promiscuous Baeyer\u2010Villiger Monooxygenase from a Thermophilic Microorganism"],"prefix":"10.1002","volume":"25","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3501-2776","authenticated-orcid":false,"given":"Amir R.","family":"Bunyat\u2010zada","sequence":"first","affiliation":[{"name":"Department of Chemistry Queen's University  Kingston, Ontario K7L 3N6 Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0001-6093-2585","authenticated-orcid":false,"given":"Stephan E.","family":"Ducharme","sequence":"additional","affiliation":[{"name":"Department of Chemistry Queen's University  Kingston, Ontario K7L 3N6 Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1923-0845","authenticated-orcid":false,"given":"Maria E.","family":"Cleveland","sequence":"additional","affiliation":[{"name":"Department of Chemistry Queen's University  Kingston, Ontario K7L 3N6 Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0009-2873-769X","authenticated-orcid":false,"given":"Esther R.","family":"Hoffman","sequence":"additional","affiliation":[{"name":"Department of Chemistry Queen's University  Kingston, Ontario K7L 3N6 Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5976-0739","authenticated-orcid":false,"given":"Graeme W.","family":"Howe","sequence":"additional","affiliation":[{"name":"Department of Chemistry Queen's University  Kingston, Ontario K7L 3N6 Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2024,9,5]]},"reference":[{"key":"e_1_2_11_1_1","doi-asserted-by":"publisher","DOI":"10.1002\/(SICI)1099-0690(199904)1999:4<737::AID-EJOC737>3.0.CO;2-B"},{"key":"e_1_2_11_2_1","doi-asserted-by":"publisher","DOI":"10.1021\/acs.chemrev.7b00167"},{"key":"e_1_2_11_3_1","doi-asserted-by":"publisher","DOI":"10.1002\/(SICI)1521-3773(19980518)37:9<1198::AID-ANIE1198>3.0.CO;2-Y"},{"key":"e_1_2_11_4_1","doi-asserted-by":"publisher","DOI":"10.1021\/acscatal.8b04132"},{"key":"e_1_2_11_5_1","doi-asserted-by":"publisher","DOI":"10.1021\/acscatal.9b03396"},{"key":"e_1_2_11_6_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jbiotec.2006.03.044"},{"key":"e_1_2_11_7_1","doi-asserted-by":"publisher","DOI":"10.1021\/op800046t"},{"key":"e_1_2_11_8_1","doi-asserted-by":"publisher","DOI":"10.1021\/acs.oprd.9b00185"},{"key":"e_1_2_11_9_1","doi-asserted-by":"publisher","DOI":"10.1002\/cbic.201000464"},{"key":"e_1_2_11_10_1","doi-asserted-by":"publisher","DOI":"10.1038\/srep28223"},{"key":"e_1_2_11_11_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jbiotec.2014.07.449"},{"key":"e_1_2_11_12_1","doi-asserted-by":"publisher","DOI":"10.1002\/bit.20869"},{"key":"e_1_2_11_13_1","doi-asserted-by":"crossref","unstructured":"S. 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