Food and Fermentation Industries >

2025 , Vol. 51 >Issue 14: 122 - 130

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.042516

Screening and identification of bacteriocin-producing strains from traditional yak fermented milk and characterization of bacteriocins

  • LIU Fengming ,
  • TAO Shengjian ,
  • LIANG Qi ,
  • FAN Ping ,
  • CHEN Xuhui ,
  • ZHAO Baotang
Expand
  • 1(College of Food Science and Engineering, Gansu Agricultural University, Lanzhou 730070, China)
    2(Functional Dairy Product Engineering Laboratory of Gansu, Lanzhou 730070, China)
    3(Lanzhou Zhuangyuan Dairy Co.Ltd., Lanzhou 730020, China)

Received date: 2025-02-25

  Revised date: 2025-03-20

  Online published: 2025-08-01

Fold

Abstract

To screen for strains with broad-spectrum bacteriocin synthesis capabilities, Escherichia coli and Staphylococcus aureus were used as indicator bacteria.A bacteriocin-producing strain, Lactiplantibacillus plantarum EL2, was isolated from traditional yak fermented milk in Gannan, Gansu, using the agar well diffusion method.The qPCR analysis revealed that EL2 harbors multiple bacteriocin synthesis genes.The crude extract of the bacteriocin synthesized by this strain not only exhibited the inhibitory activity against Gram-positive bacteria, which was common for bacteriocins, but also could inhibit Gram-negative bacteria and some fungi.Measurements of intracellular material leakage in E.coli and scanning electron microscopy observations indicated that the bacteriocin disrupts cell membrane integrity, leading to bacterial death.The inhibition efficiency was concentration-dependent.The bacteriocin showed good tolerance to temperature and pH variations, suggesting stable activity and broad application potential.It was sensitive to digestive proteases, lacked hemolytic activity, and exhibited no significant cytotoxicity to Caco-2 cells at low to medium concentrations, indicating good biosafety.The discovery of EL2 and its bacteriocin provides new resources and a theoretical basis for developing natural preservatives, highlighting its potential for further research and application.

Key words： bacteriocin; broad-spectrum antimicrobial activity; isolation and screening; Lactiplantibacillus plantarum; gene identification; antimicrobial mechanism; safety evaluation

Cite this article

LIU Fengming , TAO Shengjian , LIANG Qi , FAN Ping , CHEN Xuhui , ZHAO Baotang . Screening and identification of bacteriocin-producing strains from traditional yak fermented milk and characterization of bacteriocins[J]. Food and Fermentation Industries, 2025 , 51(14) : 122 -130 . DOI: 10.13995/j.cnki.11-1802/ts.042516

References

[1] ODEYEMI O A, ALEGBELEYE O O, STRATEVA M, et al. Understanding spoilage microbial community and spoilage mechanisms in foods of animal origin[J]. Comprehensive Reviews in Food Science and Food Safety, 2020, 19(2):311-331.
[2] ABD-ELHAKIM Y M, BEHAIRY A, HASHEM M M M, et al. Toll-like receptors and nuclear factor kappa B signaling pathway involvement in hepatorenal oxidative damage induced by some food preservatives in rats[J]. Scientific Reports, 2023, 13(1):5938.
[3] DONG H Y, XU Y, ZHANG Q Q, et al. Activity and safety evaluation of natural preservatives[J]. Food Research International, 2024, 190:114548.
[4] 叶庆磊, 张颖, 刘子豪, 等. 一株拮抗单增李斯特菌的蒙氏肠球菌A9-1A的筛选及其细菌素抗菌活性分析[J]. 微生物学通报, 2024, 51(12):5177-5192.
YE Q L, ZHANG Y, LIU Z H, et al. Enterococcus mundtii A9-1A with antagonistic effect on Listeria monocytogenes:Screening and characterization of antimicrobial activity of its bacteriocin[J]. Microbiology China, 2024, 51(12):5177-5192.
[5] 胡伟中, 李慧颖 ,史早,等. 抑制单增李斯特菌的益生乳酸菌筛选及原位抑制效果研究[J]. 食品与发酵工业, 2025, 51(10):23-32.
HU W Z, LI H Y, SHI Z, et al. Screening of probiotic lactic acid bacteria for inhibiting Listeria monocytogenes and study on their in situ inhibitory effects[J]. Food and Fermentation Industries, 2025, 51(10):23-32.
[6] 高雅茹, 张灏, 黄晓宇, 等. 屎肠球菌ID4产生的细菌素及作用机制研究[J]. 食品与发酵工业, 2025, 51(2):16-24.
GAO Y R, ZHANG H, HUANG X Y, et al. Studies on antibacterial activity and mechanism of Enterococcus faecium ID4 bacteriocin[J]. Food and Fermentation Industries, 2025, 51(2):16-24.
[7] WU J J, ZANG M W, WANG S W, et al. Nisin: From a structural and meat preservation perspective[J]. Food Microbiology, 2023, 111:104207.
[8] ZHU R F, LI B Y, HAN S W, et al. Purification, characterization, and antimicrobial mechanism of a novel broad-spectrum bacteriocin produced by Lactiplantibacillus plantarum SCA12 from Traditional Chinese Fermented Mustard greens[J]. LWT, 2024, 211:116933.
[9] LUO L L, YI L H, CHEN J X, et al. Antibacterial mechanisms of bacteriocin BM1157 against Escherichia coli and Cronobacter sakazakii[J]. Food Control, 2021, 123:107730.
[10] TODOROV S D, DE MELO FRANCO B G, TAGG J R. Bacteriocins of Gram-positive bacteria having activity spectra extending beyond closely-related species[J]. Beneficial Microbes, 2019, 10(3):315-328.
[11] 朱潇, 梁琪, 王湘竹, 等. 基于Illumina MiSeq高通量技术比较甘肃藏区传统牦牛发酵乳制品细菌菌群多样性[J]. 中国食品学报, 2021, 21(4): 336-344.
ZHU X, LIANG Q, WANG X Z, et al. Comparison of bacterial community diversity in traditional Tibetan yak fermented dairy products from Gansu based on Illumina MiSeq high-throughput technology [J]. Journal of Chinese Institute of Food Science and Technology, 2021, 21(4): 336-344.
[12] 郑雪, 梁琪, 宋雪梅, 等. 1株牦牛乳源产细菌素融合魏斯氏菌ZW21全基因组测序及序列分析[J]. 食品科学, 2023, 44(20): 119-126.
ZHENG X, LIANG Q, SONG X M, et al. Whole genome sequencing and sequence analysis of a bacteriocin-producing Weissella confusa strain ZW21 from yak milk [J]. Food Science, 2023, 44(20): 119-126.
[13] TANG J T, YI L H, ZENG K F. Isolation of antagonistic lactic acid bacteria from traditional fermented foods of China and biocontrol mode of Leuconostoc mesenteroides SNP037 against green pepper soft rot caused by Pectobacterium carotovorum[J]. Postharvest Biology and Technology, 2024, 212:112872.
[14] 张开屏, 赵艳红, 李权威, 等. 乳酸菌抗氧化特性及其katA基因分析[J]. 中国食品学报, 2021, 21(7):37-43.
ZHANG K P, ZHAO Y H, LI Q W, et al. Antioxidant properties of Lactobacillus and its KatA gene analysis[J]. Journal of Chinese Institute of Food Science and Technology, 2021, 21(7):37-43.
[15] MERAL-AKTAŞ H. Bacteriocin characterization of Enterococcus faecium isolates and evaluation of their in situ anti-Listerial activity in Beyaz cheese[J]. Food Bioscience, 2024, 61:104741.
[16] SOLTANI S, ZIRAH S, REBUFFAT S, et al. Gastrointestinal stability and cytotoxicity of bacteriocins from gram-positive and gram-negative bacteria: A comparative in vitro study[J]. Frontiers in Microbiology, 2022, 12:780355.
[17] ROJAS E R, BILLINGS G, ODERMATT P D, et al. The outer membrane is an essential load-bearing element in Gram-negative bacteria[J]. Nature, 2018, 559(7715):617-621.
[18] WU S B, LIU J H, LIU C J, et al. Quorum sensing for population-level control of bacteria and potential therapeutic applications[J]. Cellular and Molecular Life Sciences, 2020, 77(7):1319-1343.
[19] KAREB O, AÏDER M. Quorum sensing circuits in the communicating mechanisms of bacteria and its implication in the biosynthesis of bacteriocins by lactic acid bacteria: A review[J]. Probiotics and Antimicrobial Proteins, 2020, 12(1):5-17.
[20] 刘国荣, 刘洋硕, 聂蓉. 微生物共培养促进植物乳杆菌RX-8高效合成细菌素的调控机制[J]. 中国食品学报, 2023, 23(9):32-43.
LIU G R, LIU Y S, NIE R. Mechanism of microbial co-culture promoting efficient bacteriocin production by Lactobacillus plantarum RX-8 [J]. Journal of Chinese Institute of Food Science and Technology, 2023, 23(9): 32-43.
[21] XU Y L, YANG L L, LI P, et al. Heterologous expression of class IIb bacteriocin plantaricin JK in Lactococcus lactis[J]. Protein Expression and Purification, 2019, 159:10-16.
[22] 杨慧, 步雨珊, 刘奥, 等. 产细菌素植物乳杆菌Q7对酸奶后酸化及品质的影响[J]. 食品与发酵工业, 2020, 46(3):30-35.
YANG H, BU Y S, LIU A, et al. The effect of bacteriocin-producing Lactobacillus plantarum Q7 on post-acidification and quality of yogurt[J]. Food and Fermentation Industries, 2020, 46(3):30-35.
[23] YI L H, LI X, LUO L L, et al. A novel bacteriocin BMP11 and its antibacterial mechanism on cell envelope of Listeria monocytogenes and Cronobacter sakazakii[J]. Food Control, 2018, 91:160-169.
[24] 高娟娟, 贾丽艳, 畅盼盼, 等. 枯草芽孢杆菌细菌素A32的抑菌机理研究[J]. 中国食品学报, 2021, 21(10):56-64.
GAO J J, JIA L Y, CHANG P P, et al. Studies on the inhibition mechanism of bacteriocin A32 producing by Bacillus subtilis[J]. Journal of Chinese Institute of Food Science and Technology, 2021, 21(10):56-64.
[25] CAO S, DU R P, ZHAO F K, et al. The mode of action of bacteriocin CHQS, a high antibacterial activity bacteriocin produced by Enterococcus faecalis TG2[J]. Food Control, 2019, 96:470-478.
[26] 王嘉悦, 潘果, 钟忻桐, 等. 肠膜明串珠菌细菌素抗金黄色葡萄球菌的生物学特性分析[J]. 微生物学通报, 2024, 51(9):3562-3571.
WANG J Y, PAN G, ZHONG X T, et al. Biological characterization of a bacteriocin produced by Leuconostoc mesenteroides inhibiting Staphylococcus aureus[J]. Microbiology China, 2024, 51(9):3562-3571.
[27] AYYASH M, ABUSHELAIBI A, AL-MAHADIN S, et al. In-vitro investigation into probiotic characterisation of Streptococcus and Enterococcus isolated from camel milk[J]. Lwt, 2018, 87:478-487.
[28] KRISHNAMOORTHI R, SRINIVASH M, MAHALINGAM P U, et al. Antimicrobial, anti-biofilm, antioxidant and cytotoxic effects of bacteriocin by Lactococcus lactis strain CH3 isolated from fermented dairy products-An in vitro and in silico approach[J]. International Journal of Biological Macromolecules, 2022, 220:291-306.
Options
Outlines

/

〈 〉

Powered by Beijing Magtech Co. Ltd,.