草鱼肠道细菌素的分离纯化及活性研究  

Purification and Antibacterial Activity Assay of Bacteriocin from Intestinal Bacteria of Grass Carp

作　　者：俞春红[1] 彭宽[2] 史玉婷[1] 孙科军[1] 陈韬[1] 

机构地区：[1]湖南农业大学动物医学院,湖南长沙410128 [2]中南林业科技大学生物技术开放性中心实验室,湖南长沙410004

出　　处：《动物医学进展》2012年第10期20-24,共5页Progress In Veterinary Medicine

基　　金：湖南省教育厅优秀青年基金项目(08B036);中南林业科技大学青年科学基金项目(2008013B)

摘　　要：为了解草鱼肠道内细菌素的产生及抗菌效果,通过解剖草鱼获取其肠道黏膜附着物,在有氧和厌氧条件下以LB培养液增菌培养,通过滤纸片法检测培养液浓缩样品对于野生大肠埃希菌的抑菌活性。然后选择厌氧条件,以M9培养基增菌培养草鱼肠道微生物并以乙醇分级沉淀、凝胶过滤层析和C18反相高效液相色谱联用方法尝试分离厌氧菌混合培养液中的抑菌活性成分。结果表明,厌氧条件下获得的培养液中存在较强抑菌活性物质,但厌氧条件下对草鱼肠道微生物进行单菌落分离培养未能检出表现有明显抑菌活性的单一菌株。分离纯化结果显示抑菌活性主要源于其中亲水性较强、分子大小约为300ku的小分子物质。In order to know the production and bacteriostatic effect of the bacteriocin in intestines of grass carp,this study obtained attachments of intestine through grass carprs vivisecting, and then the intestinal microorganisms were cultured by aerobic and anaerobic cultivation with LB culture medium. The culture solution was concentrated, and the antibacterial activity to Escherichia coli by paper-disc method was test ed. Than,the multiple purifications of concentrates from anaerobic culture of total intestinal microorgan- isms were amplified through anaerobic cultivation with M9 culture medium, including ethyl alcohol precipitation, the gel filtration chromatography and RP-HPLC were used to isolate antibacterial materials. The results revealed that the concentrates from anaerobic cultivation inhibited the growth of E. coli evidently. In the anaerobic cultivation, however, the concentrates from single colony culture did not show any anti bacterial activity to E. coll. The results of the purification showed that the antimicrobials are highly hydrophilic substances with a molecular weight of about 300 ku.

关 键 词：草鱼 细菌素 抑菌活性 乙醇分级沉淀 反相高效液相色谱法 

分 类 号：Q74[生物学—分子生物学]