| 鸵鸟异嗜白细胞抗菌肽的分离纯化及部分性质研究 |
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| 引用本文： | 齐珂珂,康相涛,韩瑞丽,李国喜,孙桂荣,田亚东.鸵鸟异嗜白细胞抗菌肽的分离纯化及部分性质研究[J].农业生物技术学报,2009,17(3):455-461. |
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| 作者姓名： | 齐珂珂 康相涛 韩瑞丽 李国喜 孙桂荣 田亚东 |
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| 作者单位： | 1. 中国农业科学院北京畜牧兽医研究所;2. 河南农业大学牧医工程学院; |
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| 摘 要： | 目的：分离纯化出鸵鸟异嗜白细胞抗菌肽，并对其部分特性进行研究，为开发新一代高效肽类抗菌药提供依据。方法：使用氯化铵裂解、超声波破碎、醋酸提取、CM-Sepharose F F弱酸性阳离子交换层析和反相高效液相色谱（RP-HPLC）等方法，从鸵鸟异嗜白细胞中分离纯化出了抗菌肽，使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：在鸵鸟异嗜白细胞中存在抗菌肽，其对金黄色葡萄球菌S.aureus 1056MRSA、鸡大肠杆菌E.coli O78和白色念珠菌C.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MIC为0.065μg/mL，峰16的分子量为4012.472Da，对鸡大肠杆菌和金黄色葡萄球菌的MIC分别为3.971μg/mL和5.245μg/mL，峰24的分子量为3542.479Da，对鸡大肠杆菌和金黄色葡萄球菌的MIC分别为26.472μg/mL和21.561μg/mL。结论：本研究得到的抗菌肽与已报道的鸵鸟抗菌肽ostricacins相比显示出更强的抑菌活性，是否为同一物质需要进一步的验证。
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| 关 键 词： | 鸵鸟 异嗜白细胞 抗菌肽 分离纯化 特性 |
| 收稿时间： | 2008-9-26 |
| 修稿时间： | 2008-10-24 |
STUDIES ON ISOLATION, PURIFICATION AND PARTIAL CHARACTERS OF OSTRICH HETEROPHILS ANTIMICROBIAL PEPTIDES |
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| Abstract: | Objective: The aim of this work was to purify and to identify active antimicrobial peptides from the heterophils of ostrich blood. Method: Ostrich heterophils antimicrobial peptides were isolated by adding of 0.83% ammonium chlorid solution, sonicatingto release the heterophils granules, suspending in acetic acid and mixing overnight to extract the antimicrobial peptides. The crude extractions were obtained. Iox-exchange chromatography and reversed phase high performance liquid chromatography (RP-HPLC) were used to separate the components present in the crude extract. 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.
Result: The results showed that there were antimicrobial substances in ostrich heterophils, and there were strongly active against Escherichia.coli O78, Staphylococcus.aureus 1056MRSA and Candida.albicans ATCC10231all the bacteria strains above. The peptides were cationic moleculesand have good character of hot stabilization. Peak 6, 16 and 24 were obtained by RP-HPLC. The MIC of peak 6 against Candida.albicans ATCC10231 was 0.065μg/mL. The molecular weight of perk 16 was 4012.472Da, and its MIC against Escherichia.coli O78 and Staphylococcus.aureus 1056MRSA were 3.971μg/mL and 5.245μg/mL respectively. The molecular weight of fractions 24 was 3542.479Da, and the MIC of fraction 16 against Escherichia.coli O78 and Staphylococcus.aureus 1056MRSA were 26.472μg/mL and 21.561μg/mL respectively. Conclusion: The peptides we obtained displayed stronger activity than ostricacins reported by others. Further work need to determine whether they are same substance. |
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