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牛红细胞源抗菌肽的分离纯化与活性鉴定
胡建和1, 杭柏林1, 王 青1, 董永军1, 王三虎1, 刘兴友1
河南科技学院 动物科学学院
摘要:
【目的】 分离纯化牛红细胞中的抗菌肽(Antimicrobial peptides,AMPs),并对其体外抗菌活性进行初步检测。【方法】 以中国荷斯坦奶牛血液为材料,通过离子交换层析法和反相高效液相色谱法(RP-HPLC)纯化抗菌肽,用琼脂糖平板扩散法测定其抗菌活性,并用质谱法测定其分子质量。【结果】 牛血液红细胞粗提物经离子交换层析获得的阳离子峰有抗菌活性;阳离子峰经RP-HPLC纯化后,共得到4个峰(F1、F2、F3和F4),其对大肠杆菌均具有抗菌活性,F1峰和F3峰对金黄色葡萄球菌具有抗菌活性,仅F1峰对白色念珠菌具有抗菌活性;经质谱分析,F1峰纯化肽的分子质量为2 562.40 u。【结论】 成功地从牛红细胞中分离纯化到了AMPs;F1峰中AMPs的抗菌谱较广,抗菌活性较强。
关键词:  抗菌肽  牛红细胞  抗菌活性
DOI:
分类号:
基金项目:
Isolation,purification and activity identification of antimicrobial peptides from bovine red blood cells
Abstract:
【Objective】 Antimicrobial peptides were isolated and purified from bovine red blood cell,and their antimicrobial activities in vitro were analyzed.【Method】 Antimicrobial peptides from bovine red blood cells were isolated by ion exchange chromatography and purified by RP-HPLC.Antimicrobial activity of purified peptide was tested by agrose radial diffusion assay.Molecular mass of purified peptide which had good antimicrobial activity was determined by mass spectrography.【Result】 The cation peak acquired by ion exchange chromatography had antimicrobial activity.Four peaks acquired by RP-HPLC had the activity against E.coli,peak F1 and peak F3 had the activity against Staphylococcus aureus,only peak F1 had the activity against Candida albicans.Molecular mass of purified peptide of peak F1 was 2 562.40 u.【Conclusion】 Antimicrobial peptides were isolated and purified successfully from bovine red blood cells.Antimicrobial peptide of peak F1 had broad antibacterial spectrum and good antimicrobial activity.
Key words:  antimicrobial peptide  bovine red blood cell  antimicrobial activity