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当前位置: 首页 > 文章 > 酸马奶源发酵乳杆菌对致病性大肠杆菌O_8的体外抑菌研究 中国畜牧兽医 2018 (10) 2856-2865
Position: Home > Articles > Study on Antibacterial Mechanisim of Lactobacillus fermentum Isolated from Koumiss on Pathogenic Escherichia coli O_8 China Animal Husbandry & Veterinary Medicine 2018 (10) 2856-2865

酸马奶源发酵乳杆菌对致病性大肠杆菌O_8的体外抑菌研究

关键词：
酸马奶;发酵乳杆菌;致病性E.coli O8;抑菌作用;无细胞发酵上清液(CFS)
摘  要：
为了探究天然发酵酸马奶中发酵乳杆菌对致病性大肠杆菌O8(E.coli O8)的抑菌机理,本试验从酸马奶中分离筛选出抑菌效果最佳的菌株,对其进行16S rDNA序列鉴定,经NCBI网站BLAST对比鉴定菌种;对菌株进行培养发酵,制备无细胞发酵上清液(CFS);通过排酸、排过氧化氢(H2O2)、不同蛋白酶处理等方法初步确定CFS中的抑菌活性物质性质及其含量;采用牛津杯法和二倍稀释法确定CFS对致病性E.coli O824h生长曲线的最佳抑菌浓度;试剂盒法测定CFS对致病性E.coli O8的细胞膜和细胞壁通透性的影响。结果显示,从酸马奶中分离出22株对致病性E.coli O8有抑制作用的菌株,对抑菌作用最好的菌株进行16S rDNA序列鉴定及系统进化树分析后确定其为发酵乳杆菌属;CFS中主要的抑菌物质为蛋白,含量为399.5μg/mL;最低抑菌浓度(MIC)和最低杀菌浓度(MBC)分别为25.0和49.9μg/mL;CFS能使致病性E.coli O8的碱性磷酸酶(AKP)含量在1h内快速升高,之后呈缓慢增长趋势,且使致病菌培养液中的蛋白含量明显升高。综上所述,发酵乳杆菌CFS的主要抑菌物质为蛋白,蛋白浓度越高抑菌能力越强;CFS通过破坏或改变致病性E.coli O8细胞膜和细胞壁的通透性,使其释放出AKP和胞内蛋白,从而在短时间内起到抑制致病性E.coli O8生长的作用。
译  名：
Study on Antibacterial Mechanisim of Lactobacillus fermentum Isolated from Koumiss on Pathogenic Escherichia coli O_8
作  者：
GUO Xuemei;WANG Chunjie;SIMUJIDE;JIMISI;CUI Yinxue;AORIGELE;College of Animal Science,Inner Mongolia Agricultural University;College of Veterinary Medicine,Inner Mongolia Agricultural University;
单  位：
GUO Xuemei%WANG Chunjie%SIMUJIDE%JIMISI%CUI Yinxue%AORIGELE%College of Animal Science,Inner Mongolia Agricultural University%College of Veterinary Medicine,Inner Mongolia Agricultural University
关键词：
koumiss;;Lactobacillus fermentum;;pathogenic E.coli O8;;bacteriostasis;;cell-free fermentation supernatant(CFS)
摘  要：
In order to study the antibacterial mechanism of Lactobacillusfermentumisolated from natural fermented koumiss on pathogenic Escherichia coli O8(E.coli O8),the strains with best bacteriostasis isolated from koumiss was screened out in this study,and then 16 S rDNA sequence was identified by BLAST of NCBI website.After fermentation,the cell-free fermentation supernatant(CFS)was prepared.The properties and content of antibacterial substances in CFS were preliminarily determined by acid excretion,hydrogen peroxide excretion and different protease treatments.The best inhibitory concentration of CFS on the growth curve of pathogenic E.coli O8 within 24 hwas determined using the Oxford cup method and two fold dilution method.The influence of CFS on cell membrane and cell wall permeability of pathogenic E.coli O8 was determined by kit method.The results showed that there were 22 strains which had antibacterial effects onpathogenic E.coli O8 isolated from koumiss,and the best strain was Lactobacillus fermentumafter 16 S rDNA sequence and phylogenetic tree analysis.Protein was the main bacteriostatic substance of CFS,and its content was 399.5μg/mL.The minimum inhibitory concentration(MIC)and the minimum bactericidal concentration(MBC)were 25.0 and 49.9μg/mL,respectively.CFS could rapidly increase the AKP content of pathogenic E.coli O8 within 1 hand then increase slowly.Moreover,it could obviously increase the protein content in the culture of the pathogen.In conclusion,the protein of Lactobacillus fermentum was the antibacterial substance on pathogenic E.coli O8.And with the increase of protein concentration,the bacteriostasis ability was stronger.CFS could release AKP and intracellular proteins by destroying or altering the permeability of cell membrane and cell wall.Thus,it could inhibit the growth of pathogenic E.coli O8 in a short time.

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