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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 σ 54  directly and positively regulates heterologous expression of the oxytetracycline biosynthetic gene cluster in  E. coli . Bioinformatics analysis indicates that σ 54  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 σ 54  may be a viable method for the production of additional polyketide products.

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