Applied Microbiology and Biotechnology

, Volume 93, Issue 4, pp 1575–1583

An erythromycin process improvement using the diethyl methylmalonate-responsive (Dmr) phenotype of the Saccharopolyspora erythraea mutB strain

  • J. Mark Weber
  • William H. Cernota
  • Melissa C. Gonzalez
  • Benjamin I. Leach
  • Andrew R. Reeves
  • Roy K. Wesley
Applied genetics and molecular biotechnology

DOI: 10.1007/s00253-011-3650-3

Cite this article as:
Weber, J.M., Cernota, W.H., Gonzalez, M.C. et al. Appl Microbiol Biotechnol (2012) 93: 1575. doi:10.1007/s00253-011-3650-3

Abstract

The Saccharopolyspora erythraea mutB knockout strain, FL2281, having a block in the methylmalonyl-CoA mutase reaction, was found to carry a diethyl methylmalonate-responsive (Dmr) phenotype in an oil-based fermentation medium. The Dmr phenotype confers the ability to increase erythromycin A (erythromycin) production from 250–300% when the oil-based medium is supplemented with 15 mM levels of this solvent. Lower concentrations of the solvent stimulated proportionately less erythromycin production, while higher concentrations had no additional benefit. Although the mutB strain is phenotypically a low-level erythromycin producer, diethyl methylmalonate supplementation allowed it to produce up to 30% more erythromycin than the wild-type (control) strain—a strain that does not show the Dmr phenotype. The Dmr phenotype represents a new class of strain improvement phenotype. A theory to explain the biochemical mechanism for the Dmr phenotype is proposed. Other phenotypes found to be associated with the mutB knockout were a growth defect and hyper-pigmentation, both of which were restored to normal by exposure to diethyl methylmalonate. Furthermore, mutB fermentations did not significantly metabolize soybean oil in the presence of diethyl methylmalonate. Finally, a novel method is proposed for the isolation of additional mutants with the Dmr phenotype.

Keywords

Diethyl methylmalonateErythromycinmutBMethylmalonyl-CoA mutaseDmrSaccharopolyspora erythraea

Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • J. Mark Weber
    • 1
  • William H. Cernota
    • 1
  • Melissa C. Gonzalez
    • 1
    • 2
  • Benjamin I. Leach
    • 1
    • 3
  • Andrew R. Reeves
    • 1
    • 4
  • Roy K. Wesley
    • 1
  1. 1.Fermalogic, IncChicagoUSA
  2. 2.Department of Cellular and Molecular BiologyNorthwestern UniversityChicagoUSA
  3. 3.Department of MedicineLos Angeles County General HospitalLos AngelesUSA
  4. 4.Coskata, Inc.WarrenvilleUSA