液体培养基中天然变链素的纯化  

Purification of natural mutacin in liquid medium

作　　者：李颂[1] 杨柳[1] 许晓芳[1] 杨佰霞[2] 柳雯[1] 

机构地区：[1]安徽医科大学口腔医学院,安徽合肥230032 [2]安徽医科大学第一附属医院检验科,安徽合肥230022

出　　处：《中国微生态学杂志》2009年第10期926-929,共4页Chinese Journal of Microecology

基　　金：国家自然科学基金(30640074);安徽省教育厅重点项目基金(KJ2007A027);安徽医科大学博士基金(2004)资助

摘　　要：目的从变形链球菌临床株的液体培养基中分离纯化变链素,为进一步从分子水平研究变链素奠定基础。方法通过抑菌活性检测,从变链临床株中选择出抑菌活性较强的菌株。用氯仿抽提法从该菌株的培养液中粗提变链素,经固相萃取和反相高效液相色谱(RP-HPLC)对粗提物进行纯化。结果获得变链素活性较强的菌株"1G"。从其200ml液体培养基中粗提出变链素约15μg,经固相萃取柱洗脱,再经过RP-HPLC2次纯化,得到有抑菌活性的成分,此为纯化的变链素。结论变链素分子量小,分离提纯步骤复杂,本实验得到纯化的变链素,为下一步研究变链素的氨基酸序列和基因序列奠定了基础。Objective To separate and purify mutacin produced from liquid medium by Streptococci mutans （S. mutans） strains, and lay a foundation for further molecular biological research of mutacin. Method The antibacteria activity of 80 strains of S. mutaus were tested by the stab culture technique against Streptococcus oralis ATCC 10557. The mutacin produced by the strain ＂1G＂ was initially purified by SPE（ Solid-phase extraction） after crude extraction by chloroform. And then the active substances were purified by RP-HPLC （Reversed-phase high performance liquid chromatography） twice. Then the purified target peptide（mutacin） was collected and freeze-dried for further study. Result Roughly 15 μg of crude mutacin was extracted from 200 ml of liquid medium of the strain ＂1G＂. Then the purified mutacin was obtained through SPE and twice RP-HPLC. Conclusion The separation and purification of mutacin has been a complex procedure due to its small molecular mass. This study on the purification of mutacin may contribute to further research of mutacin.

关 键 词：变形链球菌 变链素 反相高效液相色谱 

分 类 号：R378.12[医药卫生—病原生物学]