The Nonribosomal Peptide Synthetase Enzyme DdaD Tethers Nβ-Fumaramoyl-l-2,3-diaminopropionate for Fe(II)/α-Ketoglutarate-Dependent Epoxidatio

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• 作者：Marie A. Hollenhorst ; Stefanie B. Bumpus ; Megan L. Matthews ; J. Martin Bollinger ; Jr. ; Neil L. Kelleher ; Christopher T. Walsh
• 刊名：Journal of the American Chemical Society
• 出版年：2010
• 出版时间：November 10, 2010
• 年：2010
• 卷：132
• 期：44
• 页码：15773-15781
• 全文大小：378K
• 年卷期：v.132,no.44(November 10, 2010)
• ISSN：1520-5126

文摘

The gene cluster from Pantoea agglomerans responsible for biosynthesis of the dapdiamide antibiotics encodes an adenylation−thiolation didomain protein, DdaD, and an Fe(II)/α-ketoglutarate-dependent dioxygenase homologue, DdaC. Here we show that DdaD, a nonribosomal peptide synthetase module, activates and sequesters N_β-fumaramoyl-lass="smallcaps">l-2,3-diaminopropionate as a covalently tethered thioester for subsequent oxidative modification of the fumaramoyl group. DdaC catalyzes Fe(II)- and α-ketoglutarate-dependent epoxidation of the covalently bound N_β-fumaramoyl-lass="smallcaps">l-2,3-diaminopropionyl-S-DdaD species to generate N_β-epoxysuccinamoyl-DAP (DAP = 2,3-diaminopropionate) in thioester linkage to DdaD. After hydrolytic release, N_β-epoxysuccinamoyl-DAP can be ligated to lass="smallcaps">l-valine by the ATP-dependent ligase DdaF to form the natural antibiotic N_β-epoxysuccinamoyl-DAP-Val.