The Circular Bacteriocins Gassericin A and Circularin A

Abstract

Gassericin A, a bacteriocin produced by Lactobacillus gasseri LA39, shows antibacterial activity against a number of Gram-positive food-borne pathogenic bacteria. Circularin A produced by Clostridium beijerinckii ATCC25752 is active against C. tyrobutyricum, a known cheese-spoilage bacterium. Both bacteriocins were purified to homogeneity from culture supernatants by reverse-phase chromatography and the subsequently determined amino acid sequences were used to clone the bacteriocin structural genes. Mature gassericin A and circularin A are class V circular bacteriocins comprised of 58 and 69 amino acid residues, respectively. Both bacteriocins are resistant to several peptidases and proteases, as are other cyclic bacteriocins. Heterologous expression of gassericin A in Escherichia coli was used to produce a non-cyclic mature peptide, which was shown to have a specific activity 173-fold lower than the circular molecule. The minimal region for production and secretion of active circularin A is comprised of five genes, as was deduced by heterologous gene expression in Enterococcus faecalis. Gassericin A and circularin A have limited mutual similarity in their primary sequences. Unlike most bacteriocins, including gassericin A, circularin A has a three-amino-acid-leader sequence.