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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 isolate and purify variant streptozotocin from the liquid culture medium of Streptococcus mutans, and to lay the foundation for the further study of variant streptozotocin at the molecular level. Methods The bacteriostatic activity test was used to select the strains with strong antibacterial activity from clinical strains with variable chain. The crude extract was purified by chloroform extraction from the culture broth of the strain. The crude extracts were purified by RP-HPLC using solid phase extraction and reversed-phase high performance liquid chromatography (RP-HPLC). As a result, the strain “1G ” which has a strong activity of changing the chain was obtained. From its 200ml liquid medium crude variable about 15μg, solid phase extraction column eluting, and then by RP-HPLC two purification, antibacterial activity of the ingredients, which is a purified variable chain. CONCLUSION: The variable molecular weight of streptozotocin is low and the steps of isolation and purification are complicated. The purified variant streptozotocin in this study lays the foundation for the further study on the amino acid sequence and gene sequence of the variant streptozotocin.