Food and Fermentation Industries >

2020 , Vol. 46 >Issue 18: 91 - 97

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.023940

Purification of bacteriocin from Lactobacillus casei and analysis of antibacterial characteristics

  • GAO Zhaojian ,
  • CAO Shanshan ,
  • SONG Yulin ,
  • DING Feihong ,
  • ZHAO Yifeng ,
  • JIAO Wei
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  • 1(College of Food (Biological) Engineering, Xuzhou Institute of Technology, Xuzhou 221018, China)
    2(Yangtze River Guiliu Food Suining Co., Ltd., Xuzhou 221000, China)
    3(Pizhou Golden Earth Fertilizer Co., Ltd., Xuzhou 221300, China)

Received date: 2020-03-12

  Revised date: 2020-04-24

  Online published: 2020-10-23

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Abstract

The aim of this work was to purify and characterize the bacteriocin (named as BaC21)produced by Lactobacillus casei XbC-36 and laid a theoretical foundation for the further development of BaC21 as a new food preservative. BaC21 was purified by ammonium sulphate precipitation, Sephadex G-15 column chromatography and reverse-phase high-performance liquid chromatography. Molecular weight of BaC21 was determined by tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis (Tricine-SDS-PAGE). Production of bacteriocin BaC21 in MRS medium reached the maximum yield of 1 695 AU/mL at the beginning of the stationary phase (18 h). The molecular weight of BaC21 was 4.4 kDa. Because its molecular mass was not similar to other known bacteriocins, BaC21 could be a novel bacteriocin. Then, the antimicrobial activity of BaC21 was nearly not affected by heating at 37-100 ℃ for up to 30 min and decreased but not fully suppressed when exposed to a temperature of 120 ℃ for 15 min, indicating high thermal stability of BaC21. BaC21 remained stable at pH values ranging from 2 to 8. Antimicrobial activity was also not affected by organic solvents and surfactants. And, SDS improved its antibacterial activity. BaC21 was of proteinaceous nature, in which complete inactivation of its antimicrobial activity was observed after being treated with proteases, while lipase and α-amylase exhibited no effect. In addition, BaC21 had broad antimicrobial spectra and especially for Gram-negative food-borne pathogens. It revealed a bacteriostatic mode of action against Staphylococcus aureus and Escherichia coli in culture media. In conclusion, bacteriocin BaC21 shows great potential as a food bio-preservative.

Key words： Lactobacillus casei; bacteriocin; separation and purification; antibacterial characteristics

Cite this article

GAO Zhaojian , CAO Shanshan , SONG Yulin , DING Feihong , ZHAO Yifeng , JIAO Wei . Purification of bacteriocin from Lactobacillus casei and analysis of antibacterial characteristics[J]. Food and Fermentation Industries, 2020 , 46(18) : 91 -97 . DOI: 10.13995/j.cnki.11-1802/ts.023940

References

[1] WAYAH S B, PHILIP K. Purification, characterization, mode of action, and enhanced production of Salivaricin mmaye1, a novel bacteriocin from Lactobacillus salivarius SPW1 of human gut origin[J].Electronic Journal of Biotechnology, 2018,35: 39-47.
[2] 匡珍, 李学英, 徐春霞, 等. 乳酸菌细菌素研究进展及其在水产养殖和加工中的应用[J].食品工业科技, 2019, 40(4): 292-298.
[3] 贾丽艳, 高娟娟, 荆旭, 等. 一株产细菌素细菌CGMCC 6624分离鉴定及其生物学特性[J].中国食品学报, 2019,19(12): 243-249.
[4] ZACHAROF M P, LOVITT R W. Bacteriocins produced by lactic acid bacteria a review article[J].APCBEE Procedia, 2012,2: 50-56.
[5] HU Y, LIU X, SHAN C, et al. Novel bacteriocin produced by Lactobacillus alimentarius FM-MM4 from a traditional Chinese fermented meat Nanx Wudl: Purification, identification and antimicrobial characteristics[J].Food Control, 2017,77: 290-297.
[6] 周佳. 肠球菌素SAU-2的纯化和特性研究[D]. 雅安:四川农业大学, 2011.
[7] COTTER P D, HILL C, ROSS R P. Bacteriocins: Developing innate immunity for food[J].Nature Reviews Microbiology, 2005, 3(10):777-788.
[8] SIDHU P K, NEHRA K. Bacteriocin-nanoconjugates as emerging compounds for enhancing antimicrobial activity of bacteriocins[J].Journal of King Saud University-Science, 2017, 31(4): 758-767.
[9] NAIR S V, BARANWAL G, CHATTERJEE M, et al. Antimicrobial activity of plumbagin, a naturally occurring naphthoquinone from Plumbago rosea, against Staphylococcus aureus and Candida albicans[J].International Journal of Medical Microbiology, 2016,306(4): 237-248.
[10] SMITHA K T, NISHA N, MAYA S, et al. Delivery of rifampicin-chitin nanoparticles into the intracellular compartment of polymorphonuclear leukocytes[J].International Journal of Biological Macromolecules, 2015,74: 36-43.
[11] NITHYA S, NIMAL T R, BARANWAL G, et al. Preparation, characterization and efficacy of lysostaphin-chitosan gel against Staphylococcus aureus[J].International Journal of Biological Macromolecules, 2018,110: 157-166.
[12] 陈文莹, 杨旭芹, 刘臻, 等. 烹调三种蔬菜对红酸汤中全反式番茄红素含量的影响[J].中国调味品, 2019,44(7): 1-4.
[13] AHN H, KIM J, KIM W J. Isolation and characterization of bacteriocinproducing Pediococcus acidilactici HW01 from malt and its potential to control beer spoilage lactic acid bacteria[J].Food Control, 2017,80: 59-66.
[14] YI L, DANG Y, WU J, et al. Purification and characterization of a novel bacteriocin produced by Lactobacillus crustorum MN047 isolated from koumiss from Xinjiang, China[J].Journal of Dairy Science, 2016,99(9): 7 002-7 015.
[15] WORAPRAYOTE W, PUMPUANG L, TOSUKHOWONG A, et al. Two putatively novel bacteriocins active against Gram-negative food borne pathogens produced by Weissella hellenica BCC 7293[J].Food Control, 2015,55: 176-184.
[16] PERUMAL V, VENKATESAN A. Antimicrobial, cytotoxic effect and purification of bacteriocin from vancomycin susceptible Enterococcus faecalis and its safety evaluation for probiotization[J].LWT- Food Science and Technology, 2017,78: 303-310.
[17] MIRANDA R O, CAMPOS-GALV?O M E M, NERO L A. Expression of genes associated with stress conditions by Listeria monocytogenes in interaction with nisin producer Lactococcus lactis[J].Food Research International, 2018,105: 897-904.
[18] MARTINEZ R C R, WACHSMAN M, TORRES N I, et al. Biochemical, antimicrobial and molecular characterization of a noncytotoxic bacteriocin produced by Lactobacillus plantarum ST71KS[J].Food Microbiology, 2013,34(2): 376-381.
[19] 陈一然. 产抑制革兰氏阴性菌细菌素菌株的筛选鉴定及植物乳杆菌素LPC818的纯化[D]. 合肥:安徽农业大学, 2011.
[20] PIARD J C, MURIANA P M, DESMAZEAUD M J, et al. Purification and partial characterization of lacticin 481, a lanthionine-containing bacteriocin produced by Lactococcus lactis subsp. lactis CNRZ 481[J].Applied and Environmental Microbiology, 1992, 58(1):279-284.
[21] 方佳琪. 乳酸菌细菌素LLC518的分离纯化及制备[D]. 合肥:安徽农业大学, 2011.
[22] KAKTCHAM P M, FOKO K E M, TCHABOU T M L, et al. Nisin-producing Lactococcus lactis subsp. lactis 2MT isolated from freshwater Nile tilapia in Cameroon: Bacteriocin screening, characterization, and optimization in a low-cost medium[J].Lebensmittel Wissenschaft Und Technologie, 2019,107: 272-279.
[23] XI Q, WANG J, DU R, et al. Purification and characterization of bacteriocin produced by a strain of Enterococcus faecalis TG2[J].Applied Biochemistry and Biotechnology, 2018,184(4): 1 106-1 119.
[24] AYED H B, MAALEJ H, HMIDET N, et al. Isolation and biochemical characterisation of a bacteriocin-like substance produced by Bacillus amyloliquefaciens An6[J].Journal of Global Antimicrobial Resistance, 2015,3(4): 255-261.
[25] GOH H F, PHILIP K. Isolation and mode of action of bacteriocin BacC1 produced by nonpathogenic Enterococcus faecium C1[J].Journal of Dairy Science, 2015,98(8): 5 080-5 090.
[26] 赵圣明, 赵岩岩, 马汉军. 植物乳杆菌JLA-9产细菌素的分离纯化[J].食品与发酵工业, 2017,43(6): 60-65.
[27] LV X, MA H, SUN M, et al. A novel bacteriocin DY4-2 produced by Lactobacillus plantarum from cutlassfish and its application as bio-preservative for the control of Pseudomonas fluorescens in fresh turbot (Scophthalmus maximus) fillets[J].Food Control, 2018,89: 22-31.
[28] 董雨馨, 谢远红, 张策, 等. 瑞士乳杆菌M14-1所产细菌素的分离纯化及抑菌特性分析[J].天然产物研究与开发, 2018,30(8): 1 437-1 443.
[29] BEN BRAÏEK O, CREMONESI P, MORANDI S, et al. Safety characterisation and inhibition of fungi and bacteria by a novel multiple enterocin-producing Enterococcus lactis 4CP3 strain[J].Microbial Pathogenesis, 2018,118: 32-38.
[30] 杨亚晋, 张日俊, 赵丽, 等. 两种高效抑菌乳酸菌素BSN4和BCN4的特性研究[J].微生物学杂志, 2017, 37(5): 65-70.
[31] YILDIRIM Z, BILGIN H, ISLEROGLU H, et al. Enterocin HZ produced by a wild Enterococcus faecium strain isolated from a traditional, starter-free pickled cheese[J].Journal of Dairy Research, 2014,81(2): 164-172.
[32] AHMAD V, AHMAD K, BAIG M H, et al. Efficacy of a novel bacteriocin isolated from Lysinibacillus sp. against Bacillus pumilus[J].Lebensmittel Wissenschaft Und Technologie, 2019, 102: 260-267.
[33] 罗怡, 张瀛, 刘颖, 等. 华贵栉孔扇贝肠道产细菌素乳酸菌的分离筛选[J].食品科技, 2019,44(2): 27-32;39.
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