Purification and Mode of Action of Antilisterial Bacteriocins Produced byLactobacillus pentosus SJ65 Isolated fromUttapamBatter

Abstract

Humans and lactic acid bacteria (LAB) have allied relationships for thousands of years, where LAB has been involved in food preservations as well as in preserving human health against several pathogens. This study aims at partial purification of bacteriocins from Lactobacillus pentosus SJ65 isolated from fermented Uttapam batter, a South Indian fermented food source, their characterization and mode of action. The bacteriocin was purified using various strategies such as acetone precipitation, gel permeation and hydrophobic interaction chromatography. The bacteriocin was stable at acidic and neutral pH and up to 100C and exhibited a wide spectrum of activity against clinically relevant gram-positive and gram-negative pathogens. However, Tris-tricine sodium dodecyl sulfate, native polyacrylamide gel electrophoresis and its gel half activity assay against Listeria monocytogenes showed that the purified bacteriocin is composed of two independent peptides with molecular weight of 3.9 and 1.6 kDa. PRACTICAL APPLICATIONSBacteriocins, proteinaceous antimicrobial compounds from lactic acid bacteria, are receiving attention for their contribution in food preservation. In addition, interests are created for their use against human bacterial, fungal infections and cancer treatment. To date, only nisin and pediocin have been approved by the United States Food and Drug Administration for use in food industries as preservatives and marketed worldwide. Other bacteriocins such as lactacins, plantaricins, pentocins, etc., are in their own lines of progress for several applications. Bacteriocins are advantageous being less toxic, having broad spectrum of activity and exhibiting antimicrobial activity at nanomolar concentrations. Thus, bacteriocins can be a potential drug candidate for replacing antibiotics to treat drug-resistant pathogens in the future.

Purification and partial characterization of antilisterial bacteriocin produced byPediococcus pentosaceusKJBC11 fromIdlibatter fermented withPiper betleleaves

“…The bacteriocin activity was decreased when heated at 100°C for 1 hr, but a complete loss of activity was observed on autoclaving at 121°C for 15 min. The results were similar to those of bacteriocins produced by LAB such as pediocin AcH (Ray, ), bacteriocin VJ13 from P. pentosaceus (Vidhyasagar & Jeevaratnam, ), and bacteriocin SJ65 from Lactobacillus pentosus (Saraniya & Jeevaratnam, ). Bacteriocin KJBC11 was stable against various detergents such as SDS, Tween 20, Tween 80, Triton X100, NaCl, and urea.…”

“…The bacteriocin KJBC11 also showed inhibitory activity against various Gram‐positive and Gram‐negative organisms in agreement with earlier reports (Jamuna & Jeevaratnam, ; Vidhyasagar & Jeevaratnam, ). Hence, it can be used for several biomedical applications and natural food preservation (Saraniya & Jeevaratnam, ). The minimum inhibitory concentration (MIC) values of bacteriocin KJBC11 against indicator organisms were presented in Table .…”

“…The active fractions from Sephadex G10 were pooled and sieved initially through gel permeation chromatography (GPC) column of Sephadex G25 (Sigma, Bangalore, India) medium grade (120 × 1.5 cm) and followed by Sephadex G25 superfine grade (30 × 1.2 cm) GPC column. The fractions were collected and monitored at 280 nm using UV‐vis spectrophotometer (Saraniya & Jeevaratnam, ).…”

“…The pooled active fractions from GPC was passed through a Sep‐Pak C18 cartridge (Waters, Bangalore) equilibrated with water having 0.1% TFA and eluted with 5, 10, 30, and 50% of isopropanol to obtain samples (fractions) of different polarity. The samples, after removing solvent by rota evaporator, were adjusted to pH 5.0 and subjected to antimicrobial activity assay against L. monocytogenes (Saraniya & Jeevaratnam, ).…”

Purification, Characterization, and Mode of Action of Pentocin JL-1, a Novel Bacteriocin Isolated from Lactobacillus pentosus, against Drug-Resistant Staphylococcus aureus

“…After the four-step purification, pentocin JL-1 was purified 70.7-fold at a yield of 4.7%. In previous studies, pentocin SJ-65 produced by L. pentosus SJ65 was purified 52-fold at a yield of 8% [ 33 ], bacteriocin KU24 produced by L. lactis KU24 was purified 24.58-fold [ 25 ], and plantaricin ZJ5 produced by L. plantarum ZJ5 was purified 139.5-fold at a yield of 1.7% [ 13 ].…”