Isolation of Lactobacillus salivarius 1077 (NRRL B-50053) and Characterization of Its Bacteriocin, Including the Antimicrobial Activity Spectrum

Abstract

Lactobacillus salivarius 1077 (NRRL B-50053) was isolated from poultry intestinal materials, and in vitro anti-Campylobacter jejuni activity was demonstrated. The isolate was then used for bacteriocin production and its enrichment. The protein content of the cell-free supernatant from the spent medium was precipitated by ammonium sulfate and dialyzed to produce the crude antimicrobial preparation. A typical bacteriocin-like response of sensitivity to proteolytic enzymes and resistance to lysozyme, lipase, and 100°C was observed with this preparation. The polypeptide was further purified by gel filtration, ion-exchange, and hydrophobicinteraction chromatography. Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS), Edman degradation, and isoelectrofocusing were used to characterize its 3,454-Da molecular mass, the amino acid sequence of its 37 residue components, and the isoelectric point of pI 9.1 of the bacteriocin. Bacteriocin L-1077 contained the class IIa bacteriocin signature N-terminal sequence YGNGV. MICs of bacteriocin L-1077 against 33 bacterial isolates (both Gram negative and Gram positive) ranged from 0.09 to 1.5 g/ml. Subsequently, the therapeutic benefit of bacteriocin L-1077 was demonstrated in market-age (40-to 43-day-old) broiler chickens colonized with both C. jejuni and Salmonella enterica serovar Enteritidis. Compared with untreated control birds, both C. jejuni and S. Enteritidis counts in colonized ceca were diminished by >4 log 10 and S. Enteritidis counts in both the liver and the spleen of treated birds were reduced by 6 to 8 log 10 /g compared with those in the nontreated control birds. Bacteriocin L-1077 appears to hold promise in controlling C. jejuni/S. Enteritidis among commercial broiler chickens.

Antibacterial Activity of Probiotic Bacteria Isolated From Broiler Feces and Commercial Strains

“…is reduced by L. reuteri (24). Recent studies have shown that Lactobacillus salivarius strains isolated from chicken gut could produce bacteriocins with antagonistic activity against Gram-positive bacteria (25). Morishita et al demonstrated that lactic acid bacteria are more effective for poultry when they have chicken origin (26).…”

New perspectives on antibacterial drug research

“…Among the newly described bacteriocins is L-1077, produced by Lactobacillus salivarius L-1077. MICs of this peptide determined against 33 Gram-negative and Grampositive bacterial isolates ranged from 0.09 to 1.5 µg/ml [88].…”

“…They are produced on the surface of ribosomes in microbial cells, while antibiotics are primarily secondary metabolites of the cell. Moreover, bacteriocins do not accumulate in tissues and can be effective against antibiotic-resistant pathogens [88,89]. However, bacteriocins do not have as broad a spectrum of antibacterial activity as antibiotics.…”

Characterization of bacterial antimicrobial peptides active against Campylobacter jejuni

“…The efficacy of L-1077 in broiler chickens was examined, causing large decreases in counts of both C. jejuni and Salmonella enterica serovar Enteriditis relative to untreated birds. 80 Edman sequencing of the isolate suggested that L-1077 consists of 37 amino acids, including an N-terminal YGNGV motif similar to the class IIa bacteriocins (Fig. 2E).…”

“…80 Following an isolation approach similar to that used for OR-7, the antimicrobial activity was attributed to a bacteriocin, termed L-1077. In addition to C. jejuni, this bacteriocin was reported to inhibit a particularly wide selection of Gramnegative and Gram-positive bacteria, such as Salmonella spp., Clostridium perfringens, and Escherichia coli.…”