Abstract
An antimycotic activity toward seven strains of Candida albicans was demonstrated erstwhile by a wild-type Enterococcus faecium isolated from a penguin rookery of the Antarctic region. In the present study the antimicrobial principle was purified by ion exchange and gel permeation chromatography and further was analyzed by LC-ESI-MS/MS. In the purification steps, the dialyzed concentrate and ion exchange fractions inhibited C. albicans MTCC 3958, 183, and SC 5314. However, the gel filtration purified fractions inhibited MTCC 3958 and 183. The data obtained from the LC-ESI-MS/MS indicate that the antimicrobial activity of the anti-Candida protein produced by E. faecium is facilitated by Sag A/Bb for the binding of the indicator organism's cell membrane. Partial N-terminal sequence revealed 12 N-terminal amino acid residues and its analysis shown that it belongs to the LysM motif. The nucleotide sequence of PCR-amplified product could detect 574 nucleotides of the LysM gene responsible for binding to chitin of the cell wall of Candida sp.
Highlights
Antimicrobial and antimycotic peptides are small cationic and amphipathic molecules, generally with fewer than 50 amino acids
The fractions collected from each step of purification was checked for the antimicrobial activity by cut-well agar assay using three C. albicans strains that are different in drug-resistance pattern and are from different sources
Based on the 12 amino acid residues from the N-terminal sequence determined in the previous study (Shekh and Roy, 2012), primers were designed and the PCR-amplified fragment of size 574 bp obtained (Figure 4) using the purified genomic DNA of the producer strain was sequenced; the nucleotide sequence was analyzed by BLAST using the National Center for Biotechnology Information (NCBI) search
Summary
Antimicrobial and antimycotic peptides are small cationic and amphipathic molecules, generally with fewer than 50 amino acids. These peptides are omnipresent and have been isolated from prokaryotes and eukaryotes in plants, bacteria, fungi, and animals (Zasloff, 2002; Bulet et al, 2004). Bacteriocins may play an important role in maintaining bacterial community structures (Riley and Wertz, 2002) in specific ecological niche. They have been proposed as probiotics for both the gastrointestinal and urogenital tracts (Redondo-Lopez et al, 1990). One of the main mechanisms used by the LAB to interfere with the colonization of pathogens and avoid proliferation of those potential pathogens is production of antimicrobial agents, such as organic acids, hydrogen peroxide and bacteriocins or related substances (Redondo-Lopez et al, 1990; Jack et al, 1995)
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