Purification and characterization of plantaricin A, a Lactobacillus plantarum bacteriocin whose activity depends on the action of two peptides Free

Abstract

SUMMARY: A bacteriocin, plantaricin A, has been purified to homogeneity by ammonium sulphate precipitation, binding to cation exchanger and Octyl-Sepharose, and reverse-phase chromatography. The bacteriocin activity was associated with two peptides, termed a and , which were separated upon reverse-phase chromatography. Bacteriocin activity required the complementary action of both the a and peptides. From the N-terminal end, 21 and 22 amino acid residues of a and , respectively, were sequenced. Further attempts at sequencing revealed no additional amino acid residues, suggesting that either the C terminus had been reached or that modifications in the next amino acid residue blocked the sequencing reaction. Judging from their amino acid sequence, a and may be encoded by the same gene, since a appeared to be a truncated form of . Alanine, the first amino acid residue at the N-terminal end of was not present at this position in a. Otherwise the sequences of a and appeared to be identical. The calculated molecular masses of the sequenced part of a and were 2426 and 2497 Da, respectively. The molecular masses of a and as determined by mass spectroscopy were 2687 30 and 2758 30 Da, respectively, indicating that (i) the only difference between a and was the presence of the N-terminal alanine residue in , and that (ii) in addition to the sequenced residues, two to three unidentified amino acid residues are present at the C-terminal ends of the a and peptides. Both a and may form amphiphilic a-helices, suggesting that they are pore-forming peptides that create cell membrane channels through a barrel-stave' mechanism.

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1993-09-01
2024-03-28
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