家蝇抗菌肽的分离纯化及性质研究  

PURIFICATION AND CHARACTERIZATION OF AN ANTIBACTERIAL PEPTIDE FROM HOUSE FLY,MUSCA DOMESTICA

作　　者：陈留存[1] 王金星[1] 刘瑶[1] 王来元[1] 王来城[1] 赵小凡[1] 

机构地区：[1]山东大学生命科学学院,山东济南250100

出　　处：《山东大学学报（理学版）》2001年第3期351-357,共7页Journal of Shandong University(Natural Science)

基　　金：山东省自然科学基金 (Y97D0 1 0 60 );高等学校骨干教师资助计划项目 (Y660 78)

摘　　要：采用阳离子交换法 ,分别以pH5.1的乙酸铵和 pH6 .6的甲酸铵作缓冲液进行 2次离子梯度洗脱 ,从家蝇成虫血淋巴中分离得到 1种抗菌肽CM Ⅲ .SDS PAGE电泳显示为 1条带 ,分子量约为 1 0 0 0 0 μ ,耐受高温的能力强 ,1 0 0℃处理 1 0min仍有活性 .它对革兰氏阳性菌 (金黄色葡萄球菌、白葡萄球菌 )和阴性菌 (绿脓假单胞菌、大肠杆菌、肠炎沙门氏菌 )都有抗性 ,其中对致病性金黄色葡萄球菌 6 538P抑菌活力最强 .CM Ⅲ对血细胞没有凝聚作用 .其抗菌机理尚待进一步研究 .One of the characteristics of the host defense of higher insects is the rapid and transient synthesis of a variety of potent antibacterial peptides.To date,several distinct inducible antibacterial peptides or peptide families have been totally or partially characterized.They can be grouped into four families:cecropins,insect defensins,proline rich peptides and glycine rich peptides.The injection of Escherichia coli in the abdomen of adult Musca domestica induces a potent antibacterial activity in the hemolymph.One of the antibacterial peptides has been isolated from the hemolymph of immunized M.domestica. Five peaks with antibacterial activity were separated by CM Sepharose chromatography and designated as CM Ⅰ to CM Ⅴ.In the next purification step the CM Ⅲ peak was most efficiently separated from contaminant proteins by rechromatography on CM Sepharose using a linear gradient ammonium formate from 0.05 to 0.6 mol/L.The purified CM Ⅲ gave only a single protein band by PAGE.Its molecular weight was estimated to be 10000 daltons by SDS PAGE.The peptide are potently active against Gram positive and Gram negative bacteria that were tested.The CM Ⅲ are heat stable,its activies were retained after 10 min incubation at 100℃,and it has no activity of coagulating red cells,which suggested that the peptide is not a lectin,the mechanism of antibacterial of this peptide is remained to be clarified.

关 键 词：家蝇 抗菌肽 革兰氏阴性菌 革兰氏阳性菌 阳离子交换 

分 类 号：Q516[生物学—生物化学]