POSTER PRESENTATION ABSTRACTS
Please note: All posters will be displayed in Salons 10-12.

THURSDAY, 10:15-11:00 AM


Li J, Aroutcheva A, Chikindas ML*.

Mode of action of a novel bacteriocin, lactocin 160 from vaginal Lactobacillus rhamnosus 160.

Rutgers, The State University of New Jersey tchikindas@aesop.rutgers.edu *Presenting author

Bacterial vaginosis (BV) is a microecological vaginal disease resulting from the replacement of healthy vaginal Lactobacillus by a multi-species pathogenic microflora. Although BV is not life threatening, it leads to many adverse effects including an increased rate of transmission and acquisition of HIV, endometriosis, as well as other pelvic inflammatory diseases and post-surgical infections. In pregnant women, BV is a proven reason for premature labor and delivery. Endotoxins produced by BV-associated bacteria can cause serious brain/CNS damage in developing fetuses.

Lactobacillus rhamnosus strain 160 isolated from a healthy subject's vagina produces an antimicrobial peptide (bacteriocin) lactocin 160 that kills several vaginal pathogens. The study's objective was to elucidate the mechanism of antimicrobial action of lactocin 160 using an established model, with Micrococcus luteus ATCC 10420 as a test organism. Sensitivity of M. luteus to lactocin 160 was determined by the diffusion assay. Loss of cellular ATP in the lactocin-treated cells was elucidated using a commercially-available ATP determination kit (luciferin-luciferase bioluminescence assay). Dissipation of membrane potential (Δψ) was studied using fluorophore DiSC3(5) with the fluorescence spectrum sensitive to changes in Δψ.

Lactocin 160 inhibited growth of M. luteus ATCC 10420 at a concentration of 5 µg/ml. There were no significant changes in the intracellular ATP level of M. luteus upon the addition of 20 µg/ml of lactocin 160. However, the extracellular ATP level was increased significantly. This means that the treatment of cells with lactocin 160 resulted in an efflux of ATP from inside the cells. Therefore, a partially purified lactocin 160 preparation (16 µg /ml of the bacteriocin in the sample) killed sensitive cells and dissipated 3.12 ± 0.36% of Δψ.

Lactocin 160 has a mode of action typical for bacteriocins. It disturbs the cellular membrane (Δψ dissipation) and induces ATP efflux, most likely due to the pore formation, which is a common mechanism of action for many bacteriocins.

 

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