Purification, Characterization and Antibacterial Mechanism of Plantaricin 2-1

To explore the basic properties and antibacterial mechanism of bacteriocin produced by Lactobacillus plantarum L3, the strain was sub-cultured in MRS medium, and the broth was extracted by ethyl acetate extraction, followed by purification serially through dextran gel chromatography, cation exchange...

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Main Authors: Mengyang LI, Xiaoxue CHE, Qingshen SUN
Format: Article
Language:zho
Published: The editorial department of Science and Technology of Food Industry 2025-02-01
Series:Shipin gongye ke-ji
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Online Access:http://www.spgykj.com/cn/article/doi/10.13386/j.issn1002-0306.2024030258
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author Mengyang LI
Xiaoxue CHE
Qingshen SUN
author_facet Mengyang LI
Xiaoxue CHE
Qingshen SUN
author_sort Mengyang LI
collection DOAJ
description To explore the basic properties and antibacterial mechanism of bacteriocin produced by Lactobacillus plantarum L3, the strain was sub-cultured in MRS medium, and the broth was extracted by ethyl acetate extraction, followed by purification serially through dextran gel chromatography, cation exchange chromatography and reverse phase high performance liquid chromatography (RP-HPLC), and finally identified by liquid chromatography tandem mass spectrometry (LC-MS/MS). Listeria monocytogenes and Escherichia coli were used as indicator strain and the antibacterial mechanism of the plantaricin was determined by crystal violet staining, scanning electron microscopy, infrared spectroscopy, fluorescence spectroscopy, extracellular alkaline phosphatase (AKP), ATP, and lactate dehydrogenase (LDH) content analysis. Results showed that plantaricin P2-1 was a novel bacteriocin with the molecular weight of 983.564 Da. The minimum inhibitory concentration (MIC) of P2-1 against two indicator strains was 28.5 μg/mL. Plantaricin P2-1 inhibited and/or eradicate the biofilms of two indicator strains. Additionally, plantaricin P2-1 could cause the death of indicator strains by destroying cell integrity, breaking down cytoderm and cytomembrane, as indicated by leakage of AKP, ATP and LDH. Additionally, P2-1 demonstrated the ability to bind to the genomic DNA of the indicator strains, thereby disrupting their DNA structure. These results suggested that P2-1 can cause multiple damages to indicator bacteria and ultimately lead to bacterial death. The above results lay a theoretical foundation for the development and application of P2-1 as a green biological preservative.
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spelling doaj-art-0c4c5fd514f5493c9113c6d63d614eba2025-02-08T08:06:20ZzhoThe editorial department of Science and Technology of Food IndustryShipin gongye ke-ji1002-03062025-02-0146413614610.13386/j.issn1002-0306.20240302582024030258-4Purification, Characterization and Antibacterial Mechanism of Plantaricin 2-1Mengyang LI0Xiaoxue CHE1Qingshen SUN2Engineering Research Center of Agricultural Microbiology Technology, Ministry of Education & Heilongjiang Provincial Key Laboratory of Plant Genetic Engineering and Biological Fermentation Engineering for Cold Region & Key Laboratory of Molecular Biology, Key Laboratory of Microbiology, College of Heilongjiang Province & School of Life Sciences, Heilongjiang University, Harbin 150080, ChinaEngineering Research Center of Agricultural Microbiology Technology, Ministry of Education & Heilongjiang Provincial Key Laboratory of Plant Genetic Engineering and Biological Fermentation Engineering for Cold Region & Key Laboratory of Molecular Biology, Key Laboratory of Microbiology, College of Heilongjiang Province & School of Life Sciences, Heilongjiang University, Harbin 150080, ChinaEngineering Research Center of Agricultural Microbiology Technology, Ministry of Education & Heilongjiang Provincial Key Laboratory of Plant Genetic Engineering and Biological Fermentation Engineering for Cold Region & Key Laboratory of Molecular Biology, Key Laboratory of Microbiology, College of Heilongjiang Province & School of Life Sciences, Heilongjiang University, Harbin 150080, ChinaTo explore the basic properties and antibacterial mechanism of bacteriocin produced by Lactobacillus plantarum L3, the strain was sub-cultured in MRS medium, and the broth was extracted by ethyl acetate extraction, followed by purification serially through dextran gel chromatography, cation exchange chromatography and reverse phase high performance liquid chromatography (RP-HPLC), and finally identified by liquid chromatography tandem mass spectrometry (LC-MS/MS). Listeria monocytogenes and Escherichia coli were used as indicator strain and the antibacterial mechanism of the plantaricin was determined by crystal violet staining, scanning electron microscopy, infrared spectroscopy, fluorescence spectroscopy, extracellular alkaline phosphatase (AKP), ATP, and lactate dehydrogenase (LDH) content analysis. Results showed that plantaricin P2-1 was a novel bacteriocin with the molecular weight of 983.564 Da. The minimum inhibitory concentration (MIC) of P2-1 against two indicator strains was 28.5 μg/mL. Plantaricin P2-1 inhibited and/or eradicate the biofilms of two indicator strains. Additionally, plantaricin P2-1 could cause the death of indicator strains by destroying cell integrity, breaking down cytoderm and cytomembrane, as indicated by leakage of AKP, ATP and LDH. Additionally, P2-1 demonstrated the ability to bind to the genomic DNA of the indicator strains, thereby disrupting their DNA structure. These results suggested that P2-1 can cause multiple damages to indicator bacteria and ultimately lead to bacterial death. The above results lay a theoretical foundation for the development and application of P2-1 as a green biological preservative.http://www.spgykj.com/cn/article/doi/10.13386/j.issn1002-0306.2024030258plartanicin 2-1purificationcharacterizationantibacterial mechanism
spellingShingle Mengyang LI
Xiaoxue CHE
Qingshen SUN
Purification, Characterization and Antibacterial Mechanism of Plantaricin 2-1
Shipin gongye ke-ji
plartanicin 2-1
purification
characterization
antibacterial mechanism
title Purification, Characterization and Antibacterial Mechanism of Plantaricin 2-1
title_full Purification, Characterization and Antibacterial Mechanism of Plantaricin 2-1
title_fullStr Purification, Characterization and Antibacterial Mechanism of Plantaricin 2-1
title_full_unstemmed Purification, Characterization and Antibacterial Mechanism of Plantaricin 2-1
title_short Purification, Characterization and Antibacterial Mechanism of Plantaricin 2-1
title_sort purification characterization and antibacterial mechanism of plantaricin 2 1
topic plartanicin 2-1
purification
characterization
antibacterial mechanism
url http://www.spgykj.com/cn/article/doi/10.13386/j.issn1002-0306.2024030258
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AT qingshensun purificationcharacterizationandantibacterialmechanismofplantaricin21