title: Engineering of the methylmalonyl-CoA metabolite node of Saccharopolyspora erythraea for increased erythromycin production.
abstract: Engineering of the methylmalonyl-CoA (mmCoA) metabolite node of the Saccharopolyspora erythraea wild-type strain through duplication of the mmCoA mutase (MCM) operon led to a 50% increase in erythromycin production in a high-performance oil-based fermentation medium. The MCM operon was carried on a 6.8kb DNA fragment in a plasmid which was inserted by homologous recombination into the S. erythraea chromosome. The fragment contained one uncharacterized gene, ORF1; three MCM related genes, mutA, mutB, meaB; and one gntR-family regulatory gene, mutR. Additional strains were constructed containing partial duplications of the MCM operon, as well as a knockout of ORF1. None of these strains showed any significant alteration in their erythromycin production profile. The combined results showed that increased erythromycin production only occurred in a strain containing a duplication of the entire MCM operon including mutR and a predicted stem-loop structure overlapping the 3' terminus of the mutR coding sequence.
authors.affiliation: Fermalogic Inc. Research and Development, 2201 West Campbell Park Drive, Chicago, IL 60612, US.
authors.forename: Andrew R
authors.initials: AR
authors.lastname: Reeves
authors.forename: Igor A
authors.initials: IA
authors.lastname: Brikun
authors.forename: William H
authors.initials: WH
authors.lastname: Cernota
authors.forename: Benjamin I
authors.initials: BI
authors.lastname: Leach
authors.forename: Melissa C
authors.initials: MC
authors.lastname: Gonzalez
authors.forename: J Mark
authors.initials: JM
authors.lastname: Weber
mesh_terms: D000214:Acyl Coenzyme A / Q000235:genetics* / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism*; D002876:Chromosomes, Bacterial / Q000235:genetics*; D004917:Erythromycin / Q000096:biosynthesis*; D005818:Genetic Engineering; D016366:Open Reading Frames / Q000235:genetics; D016930:Saccharopolyspora / Q000235:genetics* / Q000378:metabolism
chemical_list: D000214:Acyl Coenzyme A; D001426:Bacterial Proteins; C015357:methylmalonyl-coenzyme A; D004917:Erythromycin