title: Analysis of eryBI, eryBIII and eryBVII from the erythromycin biosynthetic gene cluster in Saccharopolyspora erythraea.
abstract: The gene cluster (ery) governing the biosynthesis of the macrolide antibiotic erythromycin A by Saccharopolyspora erythraea contains, in addition to the eryA genes encoding the polyketide synthase, two regions containing genes for later steps in the pathway. The region 5' of eryA that lies between the known genes ermE (encoding the erythromycin resistance methyltransferase) and eryBIII (encoding a putative S-adenosylmethionine-dependent methyltransferase), and that contains the gene eryBI (orf2), has now been sequenced. The inferred product of the eryBI gene shows striking sequence similarity to authentic beta-glucosidases. Specific mutants were created in eryBI, and the resulting strains were found to synthesise erythromycin A, showing that this gene, despite its position in the biosynthetic gene cluster, is not essential for erythromycin biosynthesis. A mutant in eryBIII and a double mutant in eryBI and eryBIII were obtained and the analysis of novel erythromycins produced by these strains confirmed the proposed function of EryBIII as a C-methyltransferase. Also, a chromosomal mutant was constructed for the previously sequenced ORF19 and shown to accumulate erythronolide B, as expected for an eryB mutant and consistent with its proposed role as an epimerase in dTDP-mycarose biosynthesis.
authors.affiliation: University of Cambridge, Department of Biochemistry, UK.
authors.forename: S
authors.initials: S
authors.lastname: Gaisser
authors.forename: G A
authors.initials: GA
authors.lastname: Böhm
authors.forename: M
authors.initials: M
authors.lastname: Doumith
authors.forename: M C
authors.initials: MC
authors.lastname: Raynal
authors.forename: N
authors.initials: N
authors.lastname: Dhillon
authors.forename: J
authors.initials: J
authors.lastname: Cortés
authors.forename: P F
authors.initials: PF
authors.lastname: Leadlay
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D001665:Binding Sites; D004917:Erythromycin / Q000378:metabolism*; D005800:Genes, Fungal; D008956:Models, Chemical; D008969:Molecular Sequence Data; D005810:Multigene Family; D016296:Mutagenesis; D016930:Saccharopolyspora / Q000235:genetics*
chemical_list: D004917:Erythromycin