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A study on characterization of new bacteriocin produced from a novel strain of Lactobacillus spicheri G2 isolated from Gundruk- a fermented vegetable product of North East India: A novel bacteriocin production from Lactobacillus spicheri G2.

ABSTRACT: Bacteriocin producing Lactobacillus spicheri G2, isolated from Gundruk - a traditional fermented vegetable product of North East India. L. spicheri G2 identified by morphological, biochemical techniques followed by 16S rRNA gene technique. The 16Sr RNA sequence of bacteriocin producer is registered in NCBI under accession no. JX481912. The bacteriocin producing potential of L. spicheri is being reported for the first time in the present investigation. Bacteriocin of L. spicheri G2 showed strong antagonism against food spoiling and pathogenic bacteria viz. Listeria monocytogenes, Staphlococcus aureus, Clostridium perfringens, Streptococcus mutans, Lactobacillus plantarum, Leuconostoc mesenteroides and Bacillus cereus. Bacteriocin production of L. spicheri G2 was enhanced by optimization of production time, pH of medium and incubation temperature by following one variable at a time method. Maximum bacteriocin activity (2000 AU/ml) was recorded in MRS broth at 34 h with an initial pH of 4.0 after incubating at 35 °C. The bacteriocin was purified by single step gel exclusion chromatography. Molecular weight of this novel bacteriocin was determined by SDS PAGE which was found to be 43 kDa. Purified bacteriocin was found resistant to high temperature and varied pH range but sensitive to proteolytic enzymes like trypsin and proteinase k, the characters desirable for food preservation.

SUBMITTER: Gautam N

PROVIDER: S-EPMC4554614 | biostudies-literature | 2015 Sep

REPOSITORIES: biostudies-literature

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A study on characterization of new bacteriocin produced from a novel strain of Lactobacillus spicheri G2 isolated from Gundruk- a fermented vegetable product of North East India: A novel bacteriocin production from Lactobacillus spicheri G2.

Gautam Neha N Sharma Nivedita N

Journal of food science and technology 20150113 9

Bacteriocin producing Lactobacillus spicheri G2, isolated from Gundruk - a traditional fermented vegetable product of North East India. L. spicheri G2 identified by morphological, biochemical techniques followed by 16S rRNA gene technique. The 16Sr RNA sequence of bacteriocin producer is registered in NCBI under accession no. JX481912. The bacteriocin producing potential of L. spicheri is being reported for the first time in the present investigation. Bacteriocin of L. spicheri G2 showed strong ...[more]

PMID: 26344995

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Project description:Lactic acid bacteria (LAB) isolated from various foods are important due to their potential to inhibit microorganisms, including drug-resistant bacteria. The objectives of this investigation were to isolate and identify antibacterial substances producing LAB from Hentak, a traditional fermented fish product of Manipur (North-East India), and to optimize the production of antagonistic substances present in cell free neutralized supernatant (CFNS) against enteric bacterial pathogens using the 'one factor at a time' (OFAT) method. Out of 10 LAB, the most potent bacterium producing antibacterial substances was isolated and identified as Lactobacillus pentosus strain LAP1 based upon morphological, biochemical and molecular characterization. MRS (de Man, Ragosa and Sharpe) medium was determined to provide better bactericidal activity (AU/ml) than other tested media against the indicator enteric bacteria, including Staphylococcus epidermidis MTTC 3615, Micrococcus luteus MTCC 106, Shigella flexneri MTCC 1457, Yersinia enterocolitica MTCC 840 and Proteus vulgaris MTCC 1771. The culture conditions (pH: 5, temperature: 30 °C and inoculum volume: 1 %) and medium components (carbon source: lactose and nitrogen source: ammonium chloride) were observed to be the most influential parameters of significant antagonistic activity of CFNS against the enteric pathogens. MRS medium supplemented with Tween20 effectively stimulated the yield of antibacterial substances. The CFNS of strain LAP1 exhibited sensitivity to proteolytic enzyme (pepsin) treatment and heat treatment (60 °C for 60 min, 100 °C for 30 min and 121 °C for 15 min) and lost its inhibitory properties. The CFNS was active at an acidic (pH 3.0) to neutral pH (pH 7.0) but lost its antagonistic properties at an alkaline pH. The CFNS obtained from strain LAP1 scavenges the DPPH (1,1-diphenyl-2 picrylhydrazyl) significantly in a concentration-dependent manner within the range of 8.8 ± 0.12-57.35 ± 0.1 %. The OFAT-based approach revealed the baseline for statistical optimization, the scale-up process and efficient production of CFNS by L. pentosus strain LAP1, which could be used as a potential antibacterial and free radical scavenging agent.

Project description:Bacteriocins from lactic acid bacteria (LAB) are of increasing interest in recent years due to their potential as natural preservatives against food and beverage spoilage microorganisms. In a screening study for LAB, we isolated from olives a strain, Lactobacillus plantarum NI326, with activity against the beverage-spoilage bacterium Alicyclobacillus acidoterrestris Genome sequencing of NI326 enabled the identification of a gene cluster (designated plc) encoding a putative circular bacteriocin and proteins involved in its modification, transport, and immunity. This novel bacteriocin, named plantaricyclin A (PlcA), was grouped into the circular bacteriocin subgroup II due to its high degree of similarity with other gassericin A-like bacteriocins. Purification of PlcA from the supernatant of Lb. plantarum NI326 resulted in an active peptide with a molecular mass of 5,570 Da, corresponding to that predicted from the (processed) PlcA amino acid sequence. The plc gene cluster was cloned and expressed in Lactococcus lactis NZ9000, resulting in the production of an active 5,570-Da bacteriocin in the supernatant. PlcA is believed to be produced as a 91-amino-acid precursor with a 33-amino-acid leader peptide, which is predicted to be removed, followed by joining of the N and C termini via a covalent linkage to form the mature 58-amino-acid circular bacteriocin PlcA. We report the characterization of a circular bacteriocin produced by Lb. plantarum The inhibition displayed against A. acidoterrestris highlights its potential use as a preservative in food and beverages.IMPORTANCE In this work, we describe the purification and characterization of an antimicrobial peptide, termed plantaricyclin A (PlcA), produced by a Lactobacillus plantarum strain isolated from olives. This peptide has a circular structure, and all genes involved in its production, circularization, and secretion were identified. PlcA shows antimicrobial activity against different strains, including Alicyclobacillus acidoterrestris, a common spoilage bacterium, which causes substantial economic losses in the beverage industry every year. In this study, we describe a circular antimicrobial peptide, PlcA, for a Lactobacillus plantarum strain.

Dataset Information

A study on characterization of new bacteriocin produced from a novel strain of Lactobacillus spicheri G2 isolated from Gundruk- a fermented vegetable product of North East India: A novel bacteriocin production from Lactobacillus spicheri G2.

Publications

A study on characterization of new bacteriocin produced from a novel strain of Lactobacillus spicheri G2 isolated from Gundruk- a fermented vegetable product of North East India: A novel bacteriocin production from Lactobacillus spicheri G2.

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