{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,7,30]],"date-time":"2025-07-30T11:59:32Z","timestamp":1753876772897,"version":"3.41.2"},"reference-count":41,"publisher":"Oxford University Press (OUP)","license":[{"start":{"date-parts":[[2024,1,1]],"date-time":"2024-01-01T00:00:00Z","timestamp":1704067200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/academic.oup.com\/journals\/pages\/open_access\/funder_policies\/chorus\/standard_publication_model"}],"funder":[{"DOI":"10.13039\/501100001700","name":"Ministry of Education, Culture, Sports, Science and Technology","doi-asserted-by":"publisher","award":["JP24K09882"],"award-info":[{"award-number":["JP24K09882"]}],"id":[{"id":"10.13039\/501100001700","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100012835","name":"Toho University","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100012835","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2024,1,9]]},"abstract":"<jats:title>Abstract<\/jats:title>\n               <jats:p>Macrolide antibiotics are biosynthesized via enzymatic modifications, including glycosylation, methylation, and oxidation, after the core macro-lactone ring is generated by a polyketide synthase system. This study explored the diversification of macrolides by combining biosynthetic enzymes and reports an approach to produce unnatural hybrid macrolide antibiotics. The cytochrome (CYP) P450 monooxygenase MycG exhibits bifunctional activity, catalyzing late-stage hydroxylation at C-14 followed by epoxidation at C-12\/13 during mycinamicin biosynthesis. The mycinose sugar of mycinamicin serves as a key molecular recognition element for binding to MycG. Thus, we subjected the hybrid macrolide antibiotic 23-O-mycinosyl-20-deoxo-20-dihydro-12,13-deepoxyrosamicin (IZI) to MycG, and confirmed that MycG catalyzed hydroxylation at C-22 and epoxidation at C-12\/13 in IZI. In addition, the introduction of mycinose biosynthesis-related genes and mycG into rosamicin-producing Micromonospora rosaria enabled the fermentative production of 22-hydroxylated and 12,13-epoxidized forms of IZI. Interestingly, MycG catalyzed the sequential oxidation of hydroxylation and epoxidation in mycinamicin biosynthesis, but only single reactions in IZI. These findings highlight the potential for expanding the application of the multifunctional P450 monooxygenase MycG for the production of unnatural compounds.<\/jats:p>","DOI":"10.1093\/femsle\/fnae080","type":"journal-article","created":{"date-parts":[[2024,9,29]],"date-time":"2024-09-29T02:44:18Z","timestamp":1727577858000},"source":"Crossref","is-referenced-by-count":0,"title":["Production of hybrid macrolide antibiotics by exploiting the specific substrate recognition characteristics of multifunctional cytochrome P450 enzyme MycG"],"prefix":"10.1093","volume":"371","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-5951-4511","authenticated-orcid":false,"given":"Yohei","family":"Iizaka","sequence":"first","affiliation":[{"name":"Faculty of Pharmaceutical Sciences, Toho University , 2-2-1 Miyama, Funabashi, Chiba 274-8510 ,","place":["Japan"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mari","family":"Yamada","sequence":"additional","affiliation":[{"name":"Faculty of Pharmaceutical Sciences, Toho University , 2-2-1 Miyama, Funabashi, Chiba 274-8510 ,","place":["Japan"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Suirei","family":"Koshino","sequence":"additional","affiliation":[{"name":"Faculty of Pharmaceutical Sciences, Toho University , 2-2-1 Miyama, Funabashi, Chiba 274-8510 ,","place":["Japan"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sawa","family":"Takahashi","sequence":"additional","affiliation":[{"name":"Faculty of Pharmaceutical Sciences, Toho University , 2-2-1 Miyama, Funabashi, Chiba 274-8510 ,","place":["Japan"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ryota","family":"Saito","sequence":"additional","affiliation":[{"name":"Department of Chemistry, Toho University , 2-2-1 Miyama, Funabashi, Chiba 274-8510 ,","place":["Japan"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"David H","family":"Sherman","sequence":"additional","affiliation":[{"name":"Life Sciences Institute, Department of Medicinal Chemistry, Chemistry, and Microbiology and Immunology, University of Michigan , Ann Arbor, MI 48109 ,","place":["United States"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yojiro","family":"Anzai","sequence":"additional","affiliation":[{"name":"Faculty of Pharmaceutical Sciences, Toho University , 2-2-1 Miyama, Funabashi, Chiba 274-8510 ,","place":["Japan"]}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"286","published-online":{"date-parts":[[2024,9,28]]},"reference":[{"key":"2024102116403017900_bib1","doi-asserted-by":"crossref","first-page":"307","DOI":"10.1271\/bbb.70.307","article-title":"Hydroxylation of testosterone by bacterial cytochrome P450 using the Escherichia coli expression system","volume":"70","author":"Agematu","year":"2006","journal-title":"Biosci Biotechnol Biochem"},{"key":"2024102116403017900_bib2","doi-asserted-by":"crossref","first-page":"1013","DOI":"10.1007\/s10295-009-0579-y","article-title":"Production of rosamicin derivatives in Micromonospora rosaria by introduction of D-mycinose biosynthetic gene with \u03a6C31-derived integration vector pSET152","volume":"36","author":"Anzai","year":"2009","journal-title":"J Ind Microbiol Biotechnol"},{"key":"2024102116403017900_bib3","doi-asserted-by":"crossref","first-page":"950","DOI":"10.1016\/j.chembiol.2008.07.014","article-title":"Functional analysis of MycCI and MycG, cytochrome P450 enzymes involved in biosynthesis of mycinamicin macrolide antibiotics","volume":"15","author":"Anzai","year":"2008","journal-title":"Chem Biol"},{"key":"2024102116403017900_bib4","doi-asserted-by":"crossref","first-page":"325","DOI":"10.1038\/ja.2010.38","article-title":"Isolation and characterization of 23-O-mycinosyl-20-dihydro-rosamicin: a new rosamicin analogue derived from engineered Micromonospora rosaria","volume":"63","author":"Anzai","year":"2010","journal-title":"J Antibiot"},{"key":"2024102116403017900_bib5","doi-asserted-by":"crossref","first-page":"3648","DOI":"10.1128\/AAC.06063-11","article-title":"Function of cytochrome P450 enzymes MycCI and MycG in Micromonospora griseorubida, a producer of the macrolide antibiotic mycinamicin","volume":"56","author":"Anzai","year":"2012","journal-title":"Antimicrob Agents Chemother"},{"key":"2024102116403017900_bib6","doi-asserted-by":"crossref","first-page":"582","DOI":"10.1271\/bbb.59.582","article-title":"Nucleotide sequence analysis of the carbomycin biosynthetic genes including the 3-O-acyltransferase gene from Streptomyces thermotolerans","volume":"59","author":"Arisawa","year":"1995","journal-title":"Biosci Biotechnol Biochem"},{"key":"2024102116403017900_bib7","doi-asserted-by":"crossref","first-page":"131","DOI":"10.7164\/antibiotics.36.131","article-title":"Biosynthesis of the macrolide antibiotic tylosin. 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