东北林蛙重组抗菌肽dybowskin-1ST部分理化特性和生物学活性研究  

The Physicochemical Properties and Biological Activity of Recombinant Antimicrobial Peptide-Dybowskin-1ST in Rana dybowskii

作　　者：李玉[1] 刘琦[1] 王敬之[1] 柴龙会[1] 周旋[1] 肖向红[1] 

机构地区：[1]东北林业大学,哈尔滨150040

出　　处：《野生动物学报》2016年第2期129-133,共5页CHINESE JOURNAL OF WILDLIFE

基　　金：东北林业大学大学生创新性实验计划项目(No.111025065);国家级大学生创新训练项目(No.111022519);国家级实验教学示范中心建设专项;国家自然科学基金项目(30870301);黑龙江省自然科学基金重点项目(ZJN0604-02)

摘　　要：目的：对具有广谱抑菌活性的东北林蛙（Rana dybowskii）重组抗菌肽dybowskin-1ST进行热稳定性、酸碱稳定性、局部过敏反应（Arthus反应）及溶血活性分析,为进一步摸清重组肽的理化特性及其应用提供科学依据。方法：将重组抗菌肽dybowskin-1ST分别置于不同环境温度（25~100℃）及不同酸碱环境（pH2~pH12）中处理4 h,观察其对金黄色葡萄球菌（Staphylococcus aureus）及大肠杆菌（Escherichia coli）的抑菌环直径,确定其稳定性;用不同浓度梯度的重组dybowskin-1ST进行溶血试验和局部过敏反应（Arthus反应）。结果：1.重组dybowskin-1ST在25~100℃温度及pH2~pH12环境下均有抑菌活性,其中,最适环境温度为25℃;重组dybowskin-1ST在pH3~pH7范围对Staphylococcus aureus抑菌活性较强（抑菌环直径为12.77~13.17 mm）,在pH3~pH8范围对Escherichia coli抑菌性较强（抑菌环直径为12.63~13.37 mm）。2.溶血试验结果显示,重组dybowskin-1ST对家兔红细胞的溶血率在0.14%~4.4%;小鼠Arthus氏试验观察其腹部皮下重组dybowskin-1ST注射部位红肿不明显,无出血、坏死等炎性反应。结论：实验结果显示重组dybowskin-1 ST具有较好稳定性、低溶血性及低致敏性,为其应用提供科学依据。We analyzed the thermal stability,pH stability,local allergic reaction（Arthus reaction）and hemolytic activity of one antimicrobial peptide dybowskin-1 ST in the frog Rana dybowskii which is characterized by broad-spectrum antibacterial activity.The physicochemical properties of its recombinant peptide were examined,thereby providing scientific basis for practical application.The recombinant antimicrobial peptide dybowskin-1ST was pretreated at different temperature（25-100℃） and different pH environments（pH2- pH12） for 4 h.Then its stability was determined by observing the size of inhibition zone against Staphylococcus aureus and Escherichia coli.Hemolysis assay and Arthus reaction were performed with different concentrations of recombinant dybowskin-1 ST.Result showed that recombinant dybowskin-1 ST have antibacterial activity at ambient temperatures of 25-10O℃ and pH2- pH12,wherein the optimum ambient temperature was251;.Recombinant dybowskin-1 ST holds strong antibacterial activity against Staphylococcus aureus（inhibition zone diameter of 12.77-13.17 mm）at pH3-pH7.However,it holds strong antibacterial activity（inhibition zone diameter of 12.63-13.37 mm） against Escherichia coli at pH3-pH8.Hemolysis test showed that the hemolysis rate of the recombinant dybowskin-1 ST on rabbit erythrocyte was 0.14%-4.4%.Arthus＇ s test showed no bleeding,necrosis,inflammation at the abdominal injection site of mice with recombinant dybowskin-1ST.We conclude that recombinant dybowskin-1ST has good stability,low hemolytic and hypoallergenic properties,thereby providing scientific basis for practical application.

关 键 词：东北林蛙 抗菌肽dybowskin-1ST 稳定性 局部过敏反应 溶血活性 

分 类 号：S858.94[农业科学—临床兽医学]