[猪链球菌2型溶菌酶的纯化及生物学活性分析]
[Purification and biological activities analysis of streptococcus suis Serotype 2 suilysin].
作者信息
Lv Qing-yu, Hao Huai-jie, Bi Li-li, Zheng Yu-ling, Jiang Yong-qiang, Lv Shu-xia
机构信息
College of Biological Science & Technology, Shenyang Agricultural University, Shenyang 11086, China.
出版信息
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2011 Apr;27(4):374-6.
AIM
To explore the purification methods of wild-type and recombinant suilysin and to evaluate their biological activities.
METHODS
Wild-type suilysin was purified by ammonium sulfate precipitation, anion-exchange chromatography and hydrophobic chromatography in turn, while recombinant suilysin was first refolded and purified by immobilized metal ion affinity chromatography, and further purified by Thiopropyl Sepharose 6B. The biological activities were evaluated by hemolysis test, cytotoxicity assay.
RESULTS
Both prepared wild-type and recombinant suilysin, with purify over 90%, have hemolysis activity and could injure target cells at high concentration while cholesterol could completely inhibit their activities.
CONCLUSION
Recombinant suilysin has similar biological activities with wild-type suilysin, and this work contributed to further study the functions of suilysin on pathogenesis of steptococcus suis.
目的
探索野生型和重组溶菌酶的纯化方法并评估其生物学活性。
方法
野生型溶菌酶依次通过硫酸铵沉淀、阴离子交换色谱和疏水色谱进行纯化，而重组溶菌酶首先通过固定化金属离子亲和色谱进行复性和纯化，然后用硫丙基琼脂糖凝胶6B进一步纯化。通过溶血试验、细胞毒性测定评估生物学活性。
结果
制备的野生型和重组溶菌酶纯度均超过90%，均具有溶血活性，在高浓度时可损伤靶细胞，而胆固醇可完全抑制其活性。
结论
重组溶菌酶与野生型溶菌酶具有相似的生物学活性，本研究为进一步研究溶菌酶在猪链球菌致病机制中的作用奠定了基础。
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