鸵鸟异嗜白细胞抗菌肽的分离、纯化及部分性质分析  

Isolation and Purification of Ostrich Heterophil Antimicrobial Peptides and Analysis of Its Partial Characters

作　　者：齐珂珂[1,2] 康相涛[1,3] 韩瑞丽[1] 李国喜[1] 孙桂荣[1] 田亚东[1] 

机构地区：[1]河南农业大学牧医工程学院,郑州450002 [2]中国农业科学院北京畜牧兽医研究所,北京100193 [3]河南家禽种质资源创新研究工程中心,郑州450002

出　　处：《农业生物技术学报》2009年第3期455-461,共7页Journal of Agricultural Biotechnology

基　　金：国家高技术研究与发展计划(863)项目(No.2002AA242021);国家农业科技跨越计划(No.2005EA750002);国家农业科技成果转化基金(No.2005410020580);河南省杰出人才创新基金(No.07420051009)资助

摘　　要：利用醋酸提取、CM-Sepharose FF弱酸性阳离子交换层析和反相高效液相色谱(RP-HPLC)从鸵鸟(Ostricacin)异嗜白细胞中分离纯化出抗菌肽;利用微量琼脂糖弥散法进行了抗菌活性与最小抑菌浓度(MIC)的测定,应用二级质谱(MS/MS)对抗菌肽的分子量进行测定。结果表明,在鸵鸟异嗜白细胞中存在抗菌肽,其对金黄色葡萄球菌(Staphyloccocus aureus)1056MRSA、鸡大肠杆菌(Escherichiacoli)O78和白色念珠菌(Candida albicans)ATCC10231具有较强的抑制作用;抗菌肽具有阳离子性质;热稳定性良好;经RP-HPLC纯化得到峰6、峰16和峰24,峰6对白色念珠菌的最小抑菌浓度为0.065μg/mL,峰16的分子量为4012.472D,对鸡大肠杆菌和金黄色葡萄球菌的MIC分别为3.971和5.245μg/mL,峰24的分子量为3542.479D,对鸡大肠杆菌和金黄色葡萄球菌的MIC分别为26.472和21.561μg/mL。Antimicrobial peptides were isolated from Ostrich （Ostricacin） heterophils by being suspended in acetic acid and mixed overnight. The crude extraction was obtained and separated by iox-exchange chromatography and reversed phase high performance liquid chromatography （RP-HPLC）. The radial diffusion plate assay method was used to determine antimicrobial activity and the minimum inhibitory concentration （MIC）. The molecular weight and amino acid sequence were determined by MS/MS. The results showed that there were antimicrobial substances in ostrich heterophils, and there were strongly active against Escherichia coil 078, Staphylococcus aureus 1056MRSA and Candida albicans ATCC 10231, respectively. The peptides were cationic molecules and had good character of hot stabilization. Peak 6, 16 and 24 were obtained by RP-HPLC. The MIC of peak 6 against Can dida albicans ATCC10231 was 0.065 txg/mL. The molecular weight ＆perk 16 was 4 012.472 D, and its MIC against F.scherichia coil 078 and Staphylococcus aureus 1056MRSA were 3.971 and 5.245 μg/mL, respectively. The molecular weight of fractions 24 was 3 542.479 D, and the MIC of fraction 16 against E. coli 078 and Staphylococcus aureus 1056MRSA were 26.472 and 21.561 μg/mL, respectively.

关 键 词：鸵鸟 异嗜白细胞 抗菌肽 分离纯化 特性 

分 类 号：S188[农业科学—农业基础科学]