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中国精品科技期刊2020
吕欣然, 李莹, 马欢欢, 缪璐欢, 杜静芳, 白凤翎, 李春, 励建荣. 辽西传统发酵食品中抗单增李斯特菌乳酸菌的筛选与鉴定[J]. 食品工业科技, 2016, (03): 143-148. DOI: 10.13386/j.issn1002-0306.2016.03.022
引用本文: 吕欣然, 李莹, 马欢欢, 缪璐欢, 杜静芳, 白凤翎, 李春, 励建荣. 辽西传统发酵食品中抗单增李斯特菌乳酸菌的筛选与鉴定[J]. 食品工业科技, 2016, (03): 143-148. DOI: 10.13386/j.issn1002-0306.2016.03.022
LV Xin- ran, LI Ying, MA Huan- huan, MIAO Lu- huan, DU Jing- fang, BAI Feng- ling, LI Chun, LI Jian-rong. Screening and identification of lactic acid bacteria with anti- Listeria monocytogenes from traditional fermented food in western Liaoning province[J]. Science and Technology of Food Industry, 2016, (03): 143-148. DOI: 10.13386/j.issn1002-0306.2016.03.022
Citation: LV Xin- ran, LI Ying, MA Huan- huan, MIAO Lu- huan, DU Jing- fang, BAI Feng- ling, LI Chun, LI Jian-rong. Screening and identification of lactic acid bacteria with anti- Listeria monocytogenes from traditional fermented food in western Liaoning province[J]. Science and Technology of Food Industry, 2016, (03): 143-148. DOI: 10.13386/j.issn1002-0306.2016.03.022

辽西传统发酵食品中抗单增李斯特菌乳酸菌的筛选与鉴定

Screening and identification of lactic acid bacteria with anti- Listeria monocytogenes from traditional fermented food in western Liaoning province

  • 摘要: 目的:从传统辽西发酵食品中筛选对单增李斯特菌具有良好拮抗作用的乳酸菌菌株。方法:采用双层琼脂扩散法筛选乳酸菌优良菌株。采用酸性实验、热处理实验、蛋白酶敏感性实验分析拮抗特性,利用生长曲线分析拮抗物质形成过程,扫描电镜分析细胞结构完整性。通过生理生化实验和16S rRNA序列进行菌株鉴定。结果:从13株乳酸菌中筛选出1株抗单增李斯特菌活性较强的菌株DL3,抗菌物质存在于无细胞上清液中。经胃蛋白酶处理后抑菌活性丧失了27.50%,胰蛋白酶和木瓜蛋白酶可使抑菌活性完全丧失,经121℃处理15 min后,抑菌活性仍保留96.88%,在p H2.5~6.5时具有抑菌活性,表明菌株DL3产生的抑菌物质可能为细菌素。添加菌株DL3无细胞上清液可使单增李斯特菌的生长曲线延迟期和稳定期延长4 h,显著地缩短了其生长期。扫描电镜结果表明经菌株DL3无细胞上清液处理的单增李斯特菌菌体变小且边缘模糊不清,其中一端细胞原生质发生泄漏。经鉴定菌株DL3为植物乳杆菌。结论:获得1株对单增李斯特菌有较强拮抗活性的植物乳杆菌DL3,该菌的拮抗活性物质可能为细菌素,可作为食品防腐剂潜在的应用菌株。 

     

    Abstract: Objective: To isolate lactic acid bacteria( LAB) from traditional fermented foods in western Liaoning province,with outstanding inhibitory activity against Listeria monocytogenes. Methods: LAB with anti-L. monocytogenes were screened by double agar diffusion methods. The antibacterial substance was analyzed using acid test,heat- treated test and protease sensitivity test,the forming process of antibacterial substance was analyzed by growth curve,the integrity of cellular structure was observed by scanning electron microscope. The strains were identified by biochemical characteristics and 16 S rRNA sequence analysis. Results: Strain DL3 with outstanding inhibitory activity against L. monocytogenes was screened from 13 strains LAB,and the antibacterial substance mainly existed in cell- free supernatants( CFS).The inhibitory activity of CFS lost 27.50% by treating with pepsin,and inhibitory activity entirely lost by treating with trypsin and papain.The antibacterial activity of CFS still remained 96.88% by thermal treatment under 121 ℃ for 15 min,and was effective within p H2.5~ p H6.5.The inhibitory substance of strain DL3 produced was preliminary determined as bacteriocin. The growth curve of L.monocytogenes lag phase and stationary phase were extended four hours by mixing CFS of LAB,which significantly shorted the logarithmic growth of L.monocytogenes.Observation under scanning electron microscope revealed that cell of L. monocytogenes became smaller and edge blur,and an apparent hole at one of the cell poles with intracellular component leakage. Strain DL3 was identified as Lactobacillus plantarum. Conclusion: Lb.plantarum DL3 with outstanding inhibitory activity against L. monocytogenes was screened,and its antibacterial substance was preliminary determined as bacteriocin. The strain DL3 had great application prospects on thedevelopment of natural bio- preservative.

     

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