title: Random transposon mutagenesis of the Saccharopolyspora erythraea genome reveals additional genes influencing erythromycin biosynthesis.
abstract: A single cycle of strain improvement was performed in Saccharopolyspora erythraea mutB and 15 genotypes influencing erythromycin production were found. Genotypes generated by transposon mutagenesis appeared in the screen at a frequency of ~3%. Mutations affecting central metabolism and regulatory genes were found, as well as hydrolases, peptidases, glycosyl transferases and unknown genes. Only one mutant retained high erythromycin production when scaled-up from micro-agar plug fermentations to shake flasks. This mutant had a knockout of the cwh1 gene (SACE_1598), encoding a cell-wall-associated hydrolase. The cwh1 knockout produced visible growth and morphological defects on solid medium. This study demonstrated that random transposon mutagenesis uncovers strain improvement-related genes potentially useful for strain engineering.
authors.affiliation: Fermalogic, Research and Development, 4222 N. Ravenswood Avenue, Chicago, IL 60613, USA.
authors.forename: Andrij
authors.initials: A
authors.lastname: Fedashchin
authors.affiliation: Fermalogic, Research and Development, 4222 N. Ravenswood Avenue, Chicago, IL 60613, USA.
authors.forename: William H
authors.initials: WH
authors.lastname: Cernota
authors.affiliation: Fermalogic, Research and Development, 4222 N. Ravenswood Avenue, Chicago, IL 60613, USA.
authors.forename: Melissa C
authors.initials: MC
authors.lastname: Gonzalez
authors.affiliation: Fermalogic, Research and Development, 4222 N. Ravenswood Avenue, Chicago, IL 60613, USA.
authors.forename: Benjamin I
authors.initials: BI
authors.lastname: Leach
authors.affiliation: Fermalogic, Research and Development, 4222 N. Ravenswood Avenue, Chicago, IL 60613, USA.
authors.forename: Noelle
authors.initials: N
authors.lastname: Kwan
authors.affiliation: Fermalogic, Research and Development, 4222 N. Ravenswood Avenue, Chicago, IL 60613, USA.
authors.forename: Roy K
authors.initials: RK
authors.lastname: Wesley
authors.affiliation: Fermalogic, Research and Development, 4222 N. Ravenswood Avenue, Chicago, IL 60613, USA mark.weber@fermalogic.com.
authors.forename: J Mark
authors.initials: JM
authors.lastname: Weber
mesh_terms: D000900:Anti-Bacterial Agents / Q000096:biosynthesis*; D053898:Biosynthetic Pathways / Q000235:genetics*; D004251:DNA Transposable Elements; D004917:Erythromycin / Q000096:biosynthesis*; D005798:Genes, Bacterial; D016680:Genome, Bacterial; D060847:Metabolic Engineering / Q000379:methods; D016254:Mutagenesis, Insertional; D016930:Saccharopolyspora / Q000235:genetics* / Q000378:metabolism*
chemical_list: D000900:Anti-Bacterial Agents; D004251:DNA Transposable Elements; D004917:Erythromycin
keywords: Actinomyces; directed evolution; erythromycin; strain improvement; transposon