Purification and characterization of a bacteriocin from Lactobacillus rhamnosus L34

Ramakrishnan, S.; Kumawat, Dinesh Kumar; Kumar, Sunil; Saxena, Anil Kumar

doi:10.1007/s13213-012-0486-8

Cited by 19 publications

(13 citation statements)

References 22 publications

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“…lactis, Lactobacillus plantarum , and L. monocytogenes can be attributed to the putative bacteriocin, considering that the mentioned antibacterial spectrum and antilisterial activity are among the main features of class II bacteriocins produced by LAB (Klaenhammer, 1993; Drider et al, 2006). Our results present some overlapping features with other reports of antimicrobial activity of bacteriocins produced by L. rhamnosus strains against E. faecalis (Todorov and Dicks, 2005; Aguilar-Uscanga et al, 2013), Micrococcus luteus (Srinivasan et al, 2013), S. aureus (Sarika et al, 2010; Srinivasan et al, 2013; Jeong and Moon, 2015), L. monocytogenes (Aguilar-Uscanga et al, 2013; Srinivasan et al, 2013; Jeong and Moon, 2015) and E. coli (Todorov and Dicks, 2005). Class II bacteriocins kill bacteria by pore formation or by interfering with the integrity of the target cell membrane, inducing permeabilization and leakage of the intracellular content (Drider et al, 2006).…”

Section: Discussionsupporting

confidence: 90%

In silico Prediction, in vitro Antibacterial Spectrum, and Physicochemical Properties of a Putative Bacteriocin Produced by Lactobacillus rhamnosus Strain L156.4

Oliveira

Silveira

Monteiro³

et al. 2017

Front. Microbiol.

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A bacteriocinogenic Lactobacillus rhamnosus L156.4 strain isolated from the feces of NIH mice was identified by 16S rRNA gene sequencing and MALDI-TOF mass spectrometry. The entire genome was sequenced using Illumina, annotated in the PGAAP, and RAST servers, and deposited. Conserved genes associated with bacteriocin synthesis were predicted using BAGEL3, leading to the identification of an open reading frame (ORF) that shows homology with the L. rhamnosus GG (ATCC 53103) prebacteriocin gene. The encoded protein contains a conserved protein motif associated a structural gene of the Enterocin A superfamily. We found ORFs related to the prebacteriocin, immunity protein, ABC transporter proteins, and regulatory genes with 100% identity to those of L. rhamnosus HN001. In this study, we provide evidence of a putative bacteriocin produced by L. rhamnosus L156.4 that was further confirmed by in vitro assays. The antibacterial activity of the substances produced by this strain was evaluated using the deferred agar-spot and spot-on-the lawn assays, and a wide antimicrobial activity spectrum against human and foodborne pathogens was observed. The physicochemical characterization of the putative bacteriocin indicated that it was sensitive to proteolytic enzymes, heat stable and maintained its antibacterial activity in a pH ranging from 3 to 9. The activity against Lactobacillus fermentum, which was used as an indicator strain, was detected during bacterial logarithmic growth phase, and a positive correlation was confirmed between bacterial growth and production of the putative bacteriocin. After a partial purification from cell-free supernatant by salt precipitation, the putative bacteriocin migrated as a diffuse band of approximately 1.0–3.0 kDa by SDS-PAGE. Additional studies are being conducted to explore its use in the food industry for controlling bacterial growth and for probiotic applications.

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Section: Discussionsupporting

confidence: 90%

In silico Prediction, in vitro Antibacterial Spectrum, and Physicochemical Properties of a Putative Bacteriocin Produced by Lactobacillus rhamnosus Strain L156.4

Oliveira

Silveira

Monteiro³

et al. 2017

Front. Microbiol.

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“…Antimicrobial activity was completely lost on treatment with proteolytic enzymes like protease, proteinase K and trypsin, suggesting a proteinaceous nature (Moraes et al, 2010). On the other hand, the activity was not lost when treated with amylase and lipase, which suggests that VJ13B is neither glycosylated nor has lipid moiety (Albano et al, 2007;Srinivasan et al, 2012;Todorov and Dicks, 2009).…”

Section: Resultsmentioning

confidence: 99%

Bacteriocin activity against various pathogens produced by Pediococcus pentosaceus VJ13 isolated from Idly batter

Vidhyasagar

Jeevaratnam

2013

Biomedical Chromatography

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Bacteriocins, an antimicrobial peptide, is known to have wide spectrum antimicrobial activity against various pathogens. Because they are easily digested in the intestine, they are considered as safe and are widely used as food preservatives. Hence their purification and characterization have attracted considerable attraction, especially for those having activity against human pathogens. In this study, the bacteriocin produced by Pediococcus pentosaceus VJ13 was precipitated with cold acetone and purified by gel permeation chromatography and hydrophobic interaction chromatography. The bacteriocin exhibited antimicrobial activity against various pathogens, like Mycobacterium smegmatis, Klebsiella pneumonia, Clostridium perfringens and Staphylococcus epidermidis. The activity of bacteriocin was lost completely after treatment with protease, which revealed its proteinaceous nature. The bacteriocin was stable up to 100°C and exhibited antilisterial property which is a characteristic feature of class IIa bacteriocins. It was active within the pH range of 2-8 and stable against various chemicals and denaturants. Tricine SDS-PAGE revealed its molecular weight to be 4.0 kDa, where the corresponding activity against Listeria monocytogenes was also noted. Treatment of L. monocytogenes with bacteriocin decreased the viable cell count, and scanning electron microscope analysis revealed membrane pore formation that resulted in the release of intracellular content, suggesting its bactericidal effect.

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“…The bacteriocin showed complete loss of antibacterial activity after trypsin, protease, and proteinase K treatment (Table ) indicating its proteinaceous nature (Moraes et al, ). Conversely, the activity was retained after catalase, amylase, and lipase treatments suggesting nonassociation of the bacteriocin with carbohydrate or lipid moieties (Srinivasan et al, ). Since bacteriocin KJBC11 was easily degraded by proteases, it was considered easily digestible and suitable for human applications (Zacharof & Lovitt, ).…”

Section: Resultsmentioning

confidence: 99%

“…Lactic Acid Bacteria (LAB) are known natural fermenters and producers of antimicrobial proteins or peptides, known as bacteriocins which are gaining attention for application as biopreservatives (Cleveland, Montville, Nes, & Chikindas, ). LAB are generally recognized as safe and their metabolites are used for food preservation and health promotion because they are beneficial and nontoxic to the host (Parada, Caron, Medeiros, & Soccol, ) and accepted by consumers as naturally occurring (Jeevaratnam, Jamuna, & Bawa, ; Srinivasan, Kumawat, Kumar, & Saxena, ). The bacteriocins produced by LAB are genetically encoded antibacterial peptides, known to be active against closely related bacteria and pathogenic bacteria such as Listeria monocytogenes , Clostridium perfringens , Bacillus cereus , and Staphylococcus aureus .…”

Section: Introductionmentioning

confidence: 99%

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

Sadishkumar

Jeevaratnam

2017

J Food Biochem

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Bacteriocins are natural antimicrobial agents mainly act against closely related bacteria and at times unrelated organisms including various food spoilage and pathogenic organisms. Bacteriocins from Lactic Acid Bacteria (LAB) have been widely used for food preservation because of their safety nature. In this study, bacteriocin from Pediococcus pentosaceus KJBC11 was purified with the recovery of 15%, using cell adsorption‐desorption technique, gel permeation chromatography, hydrophobic interaction chromatography, and reversed phase HPLC successively. Molecular weight of bacteriocin KJBC11 was determined to be 4.5 kDa using Tricine SDS‐PAGE. The bacteriocin KJBC11 showed strong inhibitory activity against various Gram‐positive and Gram‐negative pathogens. Bacteriocin KJBC11 activity was not altered when treated with amylase, lipase, and trypsin but inactivated by protease and proteinase enzyme treatment. It was heat stable (100°C, 1 hr) and exhibited strong antimicrobial activity within the pH 3–7. Its mode of action was bactericidal in nature as revealed against Listeria monocytogenes showing leakage of internal contents out of the cell. Bacteriocin KJBC11 could belong to Class IIa‐pediocin like bacteriocins based on its characteristics as well as the presence of pediocin gene in the genome of the isolate KJBC11. Practical applications Studies on Lactic Acid Bacteria and bacteriocin have attracted increasing attention for a long time, due to their established role in food fermentation and preservation. The antimicrobial properties of bacteriocin became a trademark approach to achieve food safety and to counter the menace of antibiotic resistant pathogens. This study revealed a potent bacteriocin with wide range of antibacterial activity against various foodborne and clinically important pathogens to have potential application in food preservation and biomedicine.

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Purification and characterization of a bacteriocin from Lactobacillus rhamnosus L34

Cited by 19 publications

References 22 publications

In silico Prediction, in vitro Antibacterial Spectrum, and Physicochemical Properties of a Putative Bacteriocin Produced by Lactobacillus rhamnosus Strain L156.4

In silico Prediction, in vitro Antibacterial Spectrum, and Physicochemical Properties of a Putative Bacteriocin Produced by Lactobacillus rhamnosus Strain L156.4

Bacteriocin activity against various pathogens produced by Pediococcus pentosaceus VJ13 isolated from Idly batter

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

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