AI Article Synopsis
- The study focused on expressing enterolysin A (EnlA), a bacteriocin with anti-listerial properties from Enterococcus faecalis, in Escherichia coli.
- The EnlA gene was cloned into a specific expression vector, resulting in the production of a functional His-tagged protein that was purified and confirmed to be active.
- However, modifications to the protein, such as expressing only parts of EnlA or deleting a section of its C-terminal domain, resulted in non-active proteins.
Article Abstract
The heterologous expression of enterolysin A (EnlA), heat-labile class III bacteriocin from Enterococcus faecalis II/1 with anti-listerial activity, was studied in Escherichia coli. The PCR amplified products of enterolysin A structural gene, N-terminal part of EnlA with endopeptidase-like activity and C-terminal part of EnlA similar to a lysis gene of bacteriophage, were cloned in prelinearized pQE-30UA expression vector. The expression of EnlA structural gene led to the synthesis and secretion of functional-active His-tagged enterolysin A protein, which was purified to homogeneity using His-Select Cartridge and was shown to be fully active against the indicator strain. The expression of N-terminal or C-terminal part of EnlA and deletion of last 58 amino acids from C-terminal domain of EnlA led to the synthesis of biologically non-active proteins.
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| http://dx.doi.org/10.1016/j.pep.2008.03.006 | DOI Listing |
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