title: Characterization of the Two Methylation Steps Involved in the Biosynthesis of Mycinose in Tylosin.
abstract: The S-adenosyl-l-methionine-dependent O-methyltransferases TylE and TylF catalyze the last two methylation reactions in the tylosin biosynthetic pathway of Streptomyces fradiae. It has long been known that the TylE-catalyzed C2‴-O-methylation of the 6-deoxy-d-allose bound to demethylmacrocin or demethyllactenocin precedes the TylF-catalyzed C3‴-O-methylation of the d-javose (C2‴-O-methylated 6-deoxy-d-allose) attached to macrocin or lactenocin. This study reveals the unexpected substrate promiscuity of TylE and TylF responsible for the biosynthesis of d-mycinose (C3‴-O-methylated d-javose) in tylosin through the identification of a new minor intermediate 2‴-O-demethyldesmycosin (2; 3‴-methyl-demethyllactenocin), which lacks a 2‴-O-methyl group on the mycinose moiety of desmycosin, along with 2‴-O-demethyltylosin (1; 3‴-methyl-demethylmacrocin) that was previously detected from the S. fradiae mutant containing a mutation in the tylE gene. These results unveil the unique substrate flexibility of TylE and TylF and demonstrate their potential for the engineered biosynthesis of novel glycosylated macrolide derivatives.
authors.affiliation: Department of Chemistry and Nanoscience, Ewha Womans University , Seoul 03760, Republic of Korea.
authors.forename: Eunji
authors.initials: E
authors.lastname: Kim
authors.affiliation: Department of Chemistry and Nanoscience, Ewha Womans University , Seoul 03760, Republic of Korea.
authors.forename: Myoung Chong
authors.initials: MC
authors.lastname: Song
authors.affiliation: Department of Chemistry and Nanoscience, Ewha Womans University , Seoul 03760, Republic of Korea.
authors.forename: Myoun Su
authors.initials: MS
authors.lastname: Kim
authors.affiliation: Department of Chemistry and Nanoscience, Ewha Womans University , Seoul 03760, Republic of Korea.
authors.forename: Ji Yoon
authors.initials: JY
authors.lastname: Beom
authors.affiliation: Department of Chemical Engineering, Kyung Hee University , Gyeonggi-do 17104, Republic of Korea.
authors.forename: Eun Yeol
authors.initials: EY
authors.lastname: Lee
authors.affiliation: Department of Chemical Engineering and Applied Chemistry, Chungnam National University , Daejeon 34134, Republic of Korea.
authors.forename: Dong-Myung
authors.initials: DM
authors.lastname: Kim
authors.affiliation: Department of Chemistry and Nanoscience, Ewha Womans University , Seoul 03760, Republic of Korea.
authors.forename: Sang-Jip
authors.initials: SJ
authors.lastname: Nam
authors.affiliation: Department of Chemistry and Nanoscience, Ewha Womans University , Seoul 03760, Republic of Korea.
authors.forename: Yeo Joon
authors.initials: YJ
authors.lastname: Yoon
mesh_terms: D000900:Anti-Bacterial Agents / Q000378:metabolism; D006601:Hexoses / Q000096:biosynthesis* / Q000737:chemistry; D007933:Leucomycins / Q000378:metabolism; D008745:Methylation; D008780:Methyltransferases / Q000378:metabolism*; D015394:Molecular Structure; D009154:Mutation; D012436:S-Adenosylmethionine / Q000378:metabolism; D013302:Streptomyces / Q000201:enzymology* / Q000235:genetics; D015645:Tylosin / Q000031:analogs & derivatives / Q000378:metabolism*
chemical_list: D000900:Anti-Bacterial Agents; C000617711:D-mycinose; D006601:Hexoses; D007933:Leucomycins; C052318:tylosin B; C041661:macrocin; D012436:S-Adenosylmethionine; D008780:Methyltransferases; D015645:Tylosin