Phenotypes and gene expression profiles of Saccharopolyspora erythraea rifampicin-resistant (rif) mutants affected in erythromycin production

Elisabetta Carata, Clelia Peano, Salvatore M. Tredici, Francesco Ferrari, Adelfia Talà, Giorgio Corti, Silvio Bicciato, Gianluca De Bellis, Pietro Alifano

Research output: Contribution to journalArticle

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Abstract

Background: There is evidence from previous works that bacterial secondary metabolism may be stimulated by genetic manipulation of RNA polymerase (RNAP). In this study we have used rifampicin selection as a strategy to genetically improve the erythromycin producer Saccharopolyspora erythraea. Results: Spontaneous rifampicin-resistant (rif) mutants were isolated from the parental strain NRRL2338 and two rif mutations mapping within rpoB, S444F and Q426R, were characterized. With respect to the parental strain, S444F mutants exhibited higher respiratory performance and up to four-fold higher final erythromycin yields; in contrast, Q426R mutants were slow-growing, developmental-defective and severely impaired in erythromycin production. DNA microarray analysis demonstrated that these rif mutations deeply changed the transcriptional profile of S. erythraea. The expression of genes coding for key enzymes of carbon (and energy) and nitrogen central metabolism was dramatically altered in turn affecting the flux of metabolites through erythromycin feeder pathways. In particular, the valine catabolic pathway that supplies propionyl-CoA for biosynthesis of the erythromycin precursor 6-deoxyerythronolide B was strongly up-regulated in the S444F mutants, while the expression of the biosynthetic gene cluster of erythromycin (ery) was not significantly affected. In contrast, the ery cluster was down-regulated (

Original languageEnglish
Article number18
JournalMicrobial Cell Factories
Volume8
DOIs
Publication statusPublished - Mar 30 2009

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Saccharopolyspora
Erythromycin
Rifampin
Transcriptome
Gene expression
Genes
Phenotype
Biosynthesis
Microarrays
Metabolites
RNA
Metabolism
DNA
Enzymes
Fluxes
Nitrogen
Carbon
Secondary Metabolism
Mutation
Valine

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

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Phenotypes and gene expression profiles of Saccharopolyspora erythraea rifampicin-resistant (rif) mutants affected in erythromycin production. / Carata, Elisabetta; Peano, Clelia; Tredici, Salvatore M.; Ferrari, Francesco; Talà, Adelfia; Corti, Giorgio; Bicciato, Silvio; De Bellis, Gianluca; Alifano, Pietro.

In: Microbial Cell Factories, Vol. 8, 18, 30.03.2009.

Research output: Contribution to journalArticle

Carata, Elisabetta ; Peano, Clelia ; Tredici, Salvatore M. ; Ferrari, Francesco ; Talà, Adelfia ; Corti, Giorgio ; Bicciato, Silvio ; De Bellis, Gianluca ; Alifano, Pietro. / Phenotypes and gene expression profiles of Saccharopolyspora erythraea rifampicin-resistant (rif) mutants affected in erythromycin production. In: Microbial Cell Factories. 2009 ; Vol. 8.
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