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Production, purification and characterization of bacteriocin from Lactobacillus murinus AU06 and its broad antibacterial spectrum.

ABSTRACT: OBJECTIVE: To study the production, purification and characterization of bacteriocin from Lactobacillus murinus AU06 isolated from marine sediments and its broad spectrum of inhibition against fish pathogens. METHODS: The selected strain was used in production, purification and characterized of bacteriocin. In addition, purified bacteriocin was tested for its antimicrobial activity against fish pathogens. RESULTS: In the present study, the bacteriocin production was found to be higher at 35 °C, pH 6.0 and was purified to 4.74 fold with 55. 38 U/mg of specific activity with the yield of 28.92%. The molecular weight of the purified bacteriocin was estimated as 21 kDa. The purified bacteriocin exhibited complete inactivation of antimicrobial activity when treated with proteinase K, pronase, chymotrypsin, trypsin, pepsin and papain. The purified bacteriocin exhibited broad inhibitory spectrum against both Gram positive and negative bacteria. CONCLUSIONS: It is concluded that the ability of bacteriocin in inhibiting a wide-range of pathogenic bacteria is of potential interest for food safety and may have future applications in food preservative.

SUBMITTER: Elayaraja S

PROVIDER: S-EPMC4025332 | biostudies-literature | 2014 May

REPOSITORIES: biostudies-literature

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Production, purification and characterization of bacteriocin from Lactobacillus murinus AU06 and its broad antibacterial spectrum.

Elayaraja Sivaramasamy S Annamalai Neelamegam N Mayavu Packiyam P Balasubramanian Thangavel T

Asian Pacific journal of tropical biomedicine 20140501 Suppl 1

<h4>Objective</h4>To study the production, purification and characterization of bacteriocin from Lactobacillus murinus AU06 isolated from marine sediments and its broad spectrum of inhibition against fish pathogens.<h4>Methods</h4>The selected strain was used in production, purification and characterized of bacteriocin. In addition, purified bacteriocin was tested for its antimicrobial activity against fish pathogens.<h4>Results</h4>In the present study, the bacteriocin production was found to b ...[more]

PMID: 25183102

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Project description:Lactobacillus plantarum zrx03 was a bacteriocin-producing strain isolated from infant's feces. The fermentation supernatant produced by this strain could strongly inhibit Escherichia coli JM109 ATCC 67387, Staphylococcus aureus ATCC 25923, and Listeria monocytogenes CICC 21633, in which the diameter of inhibition zone was 12.83 ± 0.62 mm, 15.08 ± 0.31 mm, 6.75 ± 0.20 mm, respectively, compared with lactic acid bacteria N1, N2, M13, M21, M31, and M37. According to amplification of 16S rRNA gene and identification of phylogenetic tree, this strain had a 1,450 bp sequence and 100% identity to the L. plantarum strain. Based on the influence of different protease treatments, such as pepsin, trypsin, papain, and proteinase K on the antimicrobial activity, this antimicrobial substance was considered to be a natural protein. Using bacteriocin produced by this strain as study object of this experiment, it had been extracted from ammonium sulfate precipitation and different organic solvents. The results showed that ethyl acetate was selected as the optimal solution to crude extraction of bacteriocin after comparing ammonium sulfate precipitation method and organic solvent extraction method, such as n-butanol, n-hexane, dichloromethane, trichloromethane, in which the diameter of the inhibition zones was above 28 mm. Results also showed the inhibition spectrum of the obtained bacteriocin had a broad spectrum of inhibition which could inhibit Gram-positive, Gram-negative, yeast. Especially, it could effectively inhibit S. aureus ATCC 25923, Bacillus subtilis CICC 10002, Bacillus anthracis CICC 20443, E. coli JM109 ATCC 67387, and Salmonella CMCC 541, and the zone diameter of inhibition has reached more than 28 mm. Moreover, it had a good thermal stability which antibacterial activity was retained 70.58% after treatment at 121°C for 30 min, and pH-stability was between pH 2.0-9.0. These results suggested bacteriocin produced by L. plantarum zrx03 had potential application prospects in food preservation.

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

Production, purification and characterization of bacteriocin from Lactobacillus murinus AU06 and its broad antibacterial spectrum.

Publications

Production, purification and characterization of bacteriocin from Lactobacillus murinus AU06 and its broad antibacterial spectrum.

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