食品与发酵工业 >

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

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

研究报告

干酪乳杆菌细菌素的分离纯化及抑菌特性分析

  • 高兆建 ,
  • 曹珊珊 ,
  • 宋玉林 ,
  • 丁飞鸿 ,
  • 赵宜峰 ,
  • 焦魏
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  • 1(徐州工程学院,食品(生物)工程学院,江苏 徐州,221018)
    2(长江桂柳食品睢宁有限公司,江苏 徐州,221000)
    3(邳州市金大地肥料有限公司,江苏 徐州,221300)
博士,副教授(本文通讯作者,E-mail:gaozhaojian@126.com)

收稿日期: 2020-03-12

  修回日期: 2020-04-24

  网络出版日期: 2020-10-23

基金资助

江苏省苏北科技计划项目(XZ-SZ201819;BC2013417;BN2015021);徐州市科技计划项目(KC17083)

收起

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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摘要

该文从干酪乳杆菌XbC-36发酵液中分离纯化细菌素(命名为BaC21)并分析抑菌特性,为开发为新型食品防腐剂奠定理论基础。硫酸铵盐析、Sephadex G-15柱层析和半制备高效液相色谱法纯化BaC21。三羟甲基氨基甘氨酸-十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis, Tricine-SDS-PAGE)法测定细菌素分子质量。菌株XbC-36在MRS培养基中发酵稳定期前期(18 h)细菌素达到最大产量1 695 AU/mL。BaC21分子质量4.4 kDa,与其他细菌素分子质量不同,推测为一种新型细菌素。BaC21在37～100 ℃加热30 min抑菌活性几乎无变化,120 ℃加热15 min抑菌活性下降,但没有完全丧失,表明具有较高热稳定性;BaC21在pH 2～8保持稳定;有机溶剂和表面活性剂对BaC21抗菌活性无影响,但SDS能提高其抗菌活性;经多种蛋白酶处理后,其抑菌活性完全失活,而脂肪酶和α-淀粉酶则无明显作用,表明BaC21具有蛋白质性质。BaC21具有广谱抑菌活性,尤其对革兰氏阴性食源性致病菌具有强抑制作用。BaC21能使金黄色葡萄球菌(Staphylococcus aureus, S. aureus)和大肠杆菌(Escherichia coli, E. coli)细胞裂解而发挥抑菌作用。BaC21优良的抑菌特性表明其有开发为食品生物防腐剂的潜力。

关键词： 干酪乳杆菌; 细菌素; 分离纯化; 抑菌特性

本文引用格式

高兆建 , 曹珊珊 , 宋玉林 , 丁飞鸿 , 赵宜峰 , 焦魏 . 干酪乳杆菌细菌素的分离纯化及抑菌特性分析[J]. 食品与发酵工业, 2020 , 46(18) : 91 -97 . DOI: 10.13995/j.cnki.11-1802/ts.023940

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

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