Organization of a cluster of erythromycin genes in Saccharopolyspora erythraea.
We used a series of gene disruptions and gene replacements to mutagenically characterize 30 kilobases of DNA in the erythromycin resistance gene (ermE) region of the Saccharopolyspora erythraea chromosome. Five previously undiscovered loci involved in the biosynthesis of erythromycin were found, eryBI, eryBII, eryCI, eryCII, and eryH; and three known loci, eryAI, eryG, and ermE, were further characterized. The new Ery phenotype, EryH, was marked by (i) the accumulation of the intermediate 6-deoxyerythronolide B (DEB), suggesting a defect in the operation of the C-6 hydroxylase system, and (ii) a block in the synthesis or addition reactions for the first sugar group. Analyses of ermE mutants indicated that ermE is the only gene required for resistance to erythromycin, and that it is not required for production of the intermediate erythronolide B (EB) or for conversion of the intermediate 3-alpha-mycarosyl erythronolide B (MEB) to erythromycin. Mutations in the eryB and eryC loci were similar to previously reported chemically induced eryB and eryC mutations blocking synthesis or attachment of the two erythromycin sugar groups. Insertion mutations in eryAI, the macrolactone synthetase, defined the largest (at least 9-kilobase) transcription unit of the cluster. These mutants help to define the physical organization of the erythromycin gene cluster, and the eryH mutants provide a source for the production of the intermediate DEB.
BioProcess Development, Abbott Laboratories, North Chicago, Illinois 60064.
J M
JM
Weber
J O
JO
Leung
G T
GT
Maine
R H
RH
Potenz
T J
TJ
Paulus
J P
JP
DeWitt
D002876:Chromosomes, Bacterial; D004352:Drug Resistance, Microbial / Q000235:genetics*; D004917:Erythromycin / Q000096:biosynthesis* / Q000494:pharmacology; D004926:Escherichia coli / Q000235:genetics; D005798:Genes, Bacterial; D008040:Genetic Linkage; D005810:Multigene Family; D009154:Mutation; D010641:Phenotype; D010957:Plasmids; D015183:Restriction Mapping; D013302:Streptomyces / Q000235:genetics*; D014169:Transformation, Bacterial
D004917:Erythromycin