屎肠球菌M-2产细菌素的纯化与特性分析  

Purification and Characterization of an Antilisteria Bacteriocin Produced by Enterococcus faecium M-2

作　　者：刘国荣[1,2] 畅晓渊[1] 吴寒宇[1] 林枫翔 李平兰[1] 

机构地区：[1]中国农业大学食品科学与营养工程学院,教育部功能乳品重点实验室,北京100083 [2]加拿大圭尔夫大学食品科学系,圭尔夫N1G2W1 [3]哈尔滨美华生物技术股份有限公司,哈尔滨150018

出　　处：《农业生物技术学报》2009年第5期925-930,共6页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金(No.30671482);国家高技术研究发展计划(863)项目(No.2006AA10A208;2006AA10Z343);“十一五”科技支撑项目(No.2006BAD04A06)资助

摘　　要：从内蒙古传统干酪中分离到的1株产细菌素屎肠球菌(Enterococcus faecium)M-2为试材,对其所产细菌素进行提取、纯化和理化特性分析。经80%硫酸铵盐析及SPSepharose Fast Flow阳离子交换树脂层析得到纯化的细菌素,其比活力可达411.204AU/mg;Tricine SDS-PAGE电泳结果显示其分子量约为3400D;特性分析发现该细菌素有很好耐热和酸碱性,经121℃处理20min后仍有活性残留,且在pH2～10的范围内活性稳定;吐温20和吐温80对其活性无影响,而SDS、司本80和尿素可导致活性损失40%～50%;蛋白酶K和胰蛋白酶处理可使其完全失活,而木瓜蛋白酶和中性蛋白酶仅可导致部分失活;871AU/mL肠球菌素M-2对单增李斯特氏菌(Listeria monocytogenes)的作用方式为杀菌;该细菌素抑菌谱较广,不仅对李斯特氏菌(Listeria spp.),葡萄球菌(Staphylococcus spp.),芽胞杆菌(Bacillus spp.)等革兰氏阳性菌有明显抑制作用,同时对大肠杆菌(Escherichiacoli)及假单胞菌(Pseudomonas spp.)等阴性菌也表现出抑菌活性,有作为食品防腐剂的良好应用前景。Purification and characterization of an antilisteria bacteriocin produced by Enterococcus faecium M-2, isolated from Inner Mongolia traditional cheese, were studied.Enterocin M-2 was purified by 80% ammonium sulphate precipitation, followed by SP-Sepharose FF cation exchange chromatography.And the specific activity of purified bacteriocin was 411.204 AU/mg.The inhibitory activity was heat-stable and still remained after 20 min at 121 ℃.The activity remained stable at pH values from 2 to 10.Tween 20 and tween 80 did not decrease the activity.However, SDS, span 80 and urea induced 40%～50% activity loss.Enterocin M-2 could be inactivated by proteinase K and trypsin, and partially inactivated by papain and neutral proteinase.Mode of action of enterocin M-2 against Listeria monocytogenes is bactericidal at 871 AU/mL.In addition, the bacteriocin had a broad inhibitory spectrum.It not only had a strong effect on gram-positive, such as Listeria spp., Staphylococcus spp., Bacillus spp.and etc, but also could inhibited several gram-negative bacteria, such as Escherichia coli, Pseudomonas spp.and etc.These results show that the bacteriocin has a potential application as food additives and bio-preservatives in food industry.

关 键 词：屎肠球菌 细菌素 纯化 特性 

分 类 号：S188[农业科学—农业基础科学]