title: Erythromycin biosynthesis. Highly efficient incorporation of polyketide chain elongation intermediates into 6-deoxyerythronolide B in an engineered Streptomyces host.
abstract: Feeding of (2S,3R)-[2,3-13C2]-2-methyl-3-hydroxypentanoyl NAC thioester (1a) to the recombinant organism Streptomyces coelicolor CH999/pCK7 harboring the complete set of eryA genes from Saccharopolyspora erythraea encoding the 6-deoxyerythronolide B synthase (DEBS) resulted in the formation of 6-deoxyerythronolide B (2a) labeled with 13C at C-12 and C-13, as evidenced by the appearance of a pair of enhanced and coupled doublets in the 13C NMR spectrum. The level of 13C enrichment was 15-20 atom% 13C, as much as 100 times higher than the usually observed efficiency of incorporation of NAC thioesters into polyketide metabolites. Similar incorporation of (2S,3R)-[3-2H,3-13C]-2-methyl-3-hydroxypentanoyl NAC thioester (1b) gave 6-deoxyerythronolide B (2b) labeled with both 13C and deuterium at C-13. The intact incorporation of both precursors confirms the normal functioning of the recombinant DEBS proteins in the heterologous host.
authors.affiliation: Department of Chemistry, Brown University, Providence, Rhode Island 02912, USA.
authors.forename: D E
authors.initials: DE
authors.lastname: Cane
authors.forename: G
authors.initials: G
authors.lastname: Luo
authors.forename: C
authors.initials: C
authors.lastname: Khosla
authors.forename: C M
authors.initials: CM
authors.lastname: Kao
authors.forename: L
authors.initials: L
authors.lastname: Katz
mesh_terms: D004274:DNA, Recombinant; D004917:Erythromycin / Q000096:biosynthesis*; D015394:Molecular Structure; D009097:Multienzyme Complexes / Q000737:chemistry / Q000378:metabolism*; D010441:Peptide Chain Elongation, Translational; D011994:Recombinant Proteins / Q000378:metabolism; D016930:Saccharopolyspora / Q000235:genetics; D013302:Streptomyces / Q000235:genetics / Q000378:metabolism*
chemical_list: D004274:DNA, Recombinant; D009097:Multienzyme Complexes; D011994:Recombinant Proteins; D004917:Erythromycin