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Metabolic Engineering of Aminocoumarins: Inactivation of the Methyltransferase Gene cloP and Generation of New Clorobiocin Derivatives in a Heterologous Host
Author(s) -
Freitag Anja,
Rapp Heike,
Heide Lutz,
Li ShuMing
Publication year - 2005
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.200500019
Subject(s) - streptomyces coelicolor , dna gyrase , gene cluster , heterologous expression , biochemistry , chemistry , streptomyces , moiety , methyltransferase , gene , antibiotics , stereochemistry , escherichia coli , biology , bacteria , recombinant dna , genetics , methylation , mutant
Aminocoumarin antibiotics are highly potent inhibitors of bacterial gyrase and represent a class of antibiotics that are very suitable for the generation of new compounds by metabolic engineering. In this study, the putative methyltransferase gene cloP in the biosynthetic gene cluster of clorobiocin was inactivated. Expression of the modified gene cluster in the heterologous host Streptomyces coelicolor M512 gave three new aminocoumarin antibiotics. The structures of the new compounds were elucidated by MS and 1 H NMR, and their antibacterial activities were determined. All three compounds lacked clorobiocin's methyl group at 4‐OH of the deoxysugar moiety, noviose. They differed from each other in the position of the 5‐methylpyrrole‐2‐carbonyl group, which was found to be attached to either 2‐OH, 3‐OH or 4‐OH of noviose. Attachment at 4‐OH resulted in the highest antibacterial activity. This is the first time that an aminocoumarin antibiotic acylated at 4‐OH in noviose has been detected.