Alternative Sigma Factor Over-Expression Enables Heterologous Expression of a Type II Polyketide Biosynthetic Pathway in Escherichia coli

David Cole Stevens, Kyle R. Conway, Nelson Pearce, Luis Roberto Villegas-Peñaranda, Anthony Garza, Christopher N. Boddy

Research output: Contribution to journalArticle

28 Citations (Scopus)

Abstract

Background:Heterologous expression of bacterial biosynthetic gene clusters is currently an indispensable tool for characterizing biosynthetic pathways. Development of an effective, general heterologous expression system that can be applied to bioprospecting from metagenomic DNA will enable the discovery of a wealth of new natural products.Methodology:We have developed a new Escherichia coli-based heterologous expression system for polyketide biosynthetic gene clusters. We have demonstrated the over-expression of the alternative sigma factor σ54 directly and positively regulates heterologous expression of the oxytetracycline biosynthetic gene cluster in E. coli. Bioinformatics analysis indicates that σ54 promoters are present in nearly 70% of polyketide and non-ribosomal peptide biosynthetic pathways.Conclusions:We have demonstrated a new mechanism for heterologous expression of the oxytetracycline polyketide biosynthetic pathway, where high-level pleiotropic sigma factors from the heterologous host directly and positively regulate transcription of the non-native biosynthetic gene cluster. Our bioinformatics analysis is consistent with the hypothesis that heterologous expression mediated by the alternative sigma factor σ54 may be a viable method for the production of additional polyketide products.

Original languageEnglish (US)
Article numbere64858
JournalPLoS One
Volume8
Issue number5
DOIs
StatePublished - May 28 2013

Fingerprint

sigma factors
Polyketides
Sigma Factor
polyketides
Biosynthetic Pathways
Multigene Family
multigene family
RNA Polymerase Sigma 54
Escherichia coli
biochemical pathways
Genes
Oxytetracycline
oxytetracycline
Bioinformatics
Computational Biology
bioinformatics
Bacterial Genes
Metagenomics
Transcription
Biological Products

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Alternative Sigma Factor Over-Expression Enables Heterologous Expression of a Type II Polyketide Biosynthetic Pathway in Escherichia coli. / Stevens, David Cole; Conway, Kyle R.; Pearce, Nelson; Villegas-Peñaranda, Luis Roberto; Garza, Anthony; Boddy, Christopher N.

In: PLoS One, Vol. 8, No. 5, e64858, 28.05.2013.

Research output: Contribution to journalArticle

Stevens, David Cole ; Conway, Kyle R. ; Pearce, Nelson ; Villegas-Peñaranda, Luis Roberto ; Garza, Anthony ; Boddy, Christopher N. / Alternative Sigma Factor Over-Expression Enables Heterologous Expression of a Type II Polyketide Biosynthetic Pathway in Escherichia coli. In: PLoS One. 2013 ; Vol. 8, No. 5.
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