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Microbiology 152 (2006), 667-673; DOI  10.1099/mic.0.28680-0
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Microbiology 152 (2006), 667-673; DOI  10.1099/mic.0.28680-0
© 2006 Society for General Microbiology

In vivo characterization of the dTDP-D-desosamine pathway of the megalomicin gene cluster from Micromonospora megalomicea

Eduardo Rodríguez1,2, Salvador Peirú1, John R. Carney2 and Hugo Gramajo1,2

1 Microbiology Division, IBR (Instituto de Biología Molecular y Celular de Rosario), Consejo Nacional de Investigaciones Científicas y Técnicas, Facultad de Ciencias, Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario, Suipacha 531, (S2002LRK) Rosario, Argentina
2 Kosan Biosciences, Inc., 3832 Bay Center Place, Hayward, CA 94545, USA

Correspondence
Hugo Gramajo
gramajo{at}ibr.gov.ar

In vivo reconstitution of the dTDP-D-desosamine pathway of the megalomicin gene cluster from Micromonospora megalomicea was achieved by expression of the genes in Escherichia coli. LC/MS/MS analysis of the dTDP-sugar intermediates produced by operons containing different sets of genes showed that production of dTDP-D-desosamine from dtdp-4-keto-6-deoxy-D-glucose requires only four biosynthetic steps, catalysed by MegCIV, MegCV, MegDII and MegDIII, and that MegCII is not involved. Instead, bioconversion studies demonstrated that MegCII is needed together with MegCIII to catalyse transfer of D-desosamine to 3-{alpha}-mycarosylerythronolide B.

Abbreviations: MEB, 3-{alpha}-mycarosylerythronolide B






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Copyright © 2006 Society for General Microbiology.