金黄色葡萄球菌细胞壁可溶性肽聚糖的分离纯化和鉴定

Isolation purification and identification of soluble peptidoglycan from staphylococcus aureus

目的从金黄色葡萄球菌中提取纯化可溶性肽聚糖并鉴定其活性。方法采用小剂量青霉素作用金黄色葡萄球菌(ATCC25923),诱导合成可溶性肽聚糖(PGN)粗品,经SephadexG-100凝胶层析进一步分离纯化;采用SLP试剂盒对纯化的可溶性PGN定性,ELISA法检测纯化的可溶性PGN对小鼠RAW264.7细胞的活化作用。结果分离纯化的可溶性PGN经SLP试剂检测阳性,该可溶性PGN可刺激RAW264.7细胞释放TNF-α,并具有明显的量效关系。结论采用青霉素诱导结合凝胶层析技术可从金黄色葡萄球菌获取纯度及生物学活性较高的可溶性PGN。

AIM：To extract and identify the soluble peptidoglycan from staphylococcus aureus. METHODS： The soluble peptidoglycan from staphylococcus aureus （ATCC25923） was released by short dose penicillin. Then, the soluble peptidoglycan was purified by sephadex G-100. The SLP reagent set was employed to identify the purified soluble peptidoglycan, and the production stimulated by the purified soluble peptidoglycan in RAW264.7 cell was observed by ELISA. RESULTS： Detected by the SLP reagent, the purified soluble peptidoglycan showed positive. The purified soluble peptidoglycan was capable of stimulating the production of TNF-α in RAW264.7 cell, and the amounts of TNF-α released were directly proportional to the concentrations of PGN. CONCLUSION： The soluble peptidoglycan from staphylococcus aureus can be obtained by penicillin and sephadex G-100, which has higher biological reactivities.