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Background  Scientific interest in  Enterococcus faecalis  has increased greatly over recent decades. Some strains are involved in food fermentation and offer health benefits, whereas others are vancomycin-resistant and cause infections that are difficult to treat. The limited availability of vectors able to express cloned genes efficiently in  E. faecalis  has hindered biotechnological studies on the bacterium’s regulatory and pathogenicity-related genes. The agmatine deiminase (AGDI) pathway of  E. faecalis , involved in the conversion of agmatine into putrescine, is driven by a response inducer gene  aguR .    Results  This study describes that the exposure to the induction factor (agmatine) results in the transcription of genes under the control of the  aguB  promoter, including the  aguBDAC  operon. A novel  E. faecalis  expression vector, named pAGEnt, combining the  aguR  inducer gene and the  aguB  promoter followed by a cloning site and a stop codon was constructed. pAGEnt was designed for the overexpression and purification of a protein fused to a 10-amino-acid His-tag at the C-terminus. The use of GFP as a reporter of gene expression in  E. faecalis  revealed that under induction with 60 mM agmatine, fluorescence reached 40 arbitrary units compared to 0 in uninduced cells.    Conclusion  pAGEnt vector can be used for the overexpression of recombinant proteins under the induction of agmatine in  E. faecalis,  with a close correlation between agmatine concentration and fluorescence when GFP was used as reporter.

An agmatine-inducible system for the expression of recombinant proteins in Enterococcus faecalis