斗米虫抗菌肽粗提液抑菌活性及其生化特性研究  

Antibacterial Activity and Biochemical Properties of Antimicrobial Peptide Crude Extract from Apriona swainsoni Larvae

作　　者：高诗琪 周卫青 苏兰 徐华潮[1] GAO Shiqi;ZHOU Weiqing;SU Lan;XU Huachao(College of Forestry and Biotechnology,Zhejiang A&F University,Hangzhou 311300,China;Chun’an County Forestry Bureau,Hangzhou City,Zhejiang Province,Hangzhou 311700,China;Lingfeng Subdistrict Office of Anji County,Huzhou City,Zhejiang Province,Huzhou 313000,China)

机构地区：[1]浙江农林大学林业与生物技术学院,杭州311300 [2]浙江省杭州市淳安县林业局,杭州311700 [3]浙江省湖州市安吉县灵峰街道办事处,湖州313000

出　　处：《动物营养学报》2023年第1期673-680,共8页CHINESE JOURNAL OF ANIMAL NUTRITION

基　　金：浙江省省院合作项目——食用蟋蟀和斗米虫培育技术优化及产品研发(2019SY02)。

摘　　要：本试验旨在测定斗米虫抗菌肽粗提液的蛋白质浓度、抑菌活性与部分生化特性,以期为斗米虫在新型抗生素筛选方面提供理论基础。用浓度均为1.5×108CFU/mL的大肠杆菌和金黄色葡萄球菌等体积混合对斗米虫开展抗菌肽诱导,分别在诱导12、24、36、48、60和72 h后提取抗菌肽,利用考马斯亮蓝法测定斗米虫抗菌肽粗提液的蛋白质浓度,采用滤纸片法测定其抑菌活性,同时对其热稳定性、反复冻融稳定性、酸碱稳定性等生化特性进行测定。结果显示:1)经大肠杆菌、金黄色葡萄球菌混合诱导的斗米虫抗菌肽粗提液的蛋白质浓度均极显著高于未诱导的对照组(P<0.01),且在诱导24 h时蛋白质浓度最高;2)斗米虫抗菌肽粗提液仅对金黄色葡萄球菌和枯草芽孢杆菌有抑制作用,对大肠杆菌和铜绿假单胞菌无抑制作用;3)生化特性的测定结果显示,斗米虫抗菌肽粗提液有较好的热稳定性和酸碱稳定性,反复冻融对其抑菌活性影响较小。综上可知,大肠杆菌、金黄色葡萄球菌等体积混合诱导斗米虫,可以产生浓度高、活性强、生化特性较稳定的抗菌肽,该研究结果可为斗米虫作为新型抗生素开发和药物筛选提供理论基础。This experiment was conducted to determine the protein concentration,antibacterial activity,and some biochemical properties of the crude extract of antimicrobial peptides from Apriona swainsoni larvae,in order to provide a theoretical basis for the screening of new antibiotics in the field of Apriona swainsoni larvae.In this experiment,Escherichia coli and Staphylococcus aureus were mixed to induce Apriona swainsoni at a concentration of 1.5×108CFU/mL,and the antimicrobial peptides were extracted after 12,24,36,48,60and 72 h of induction,respectively. The coomassie brilliant blue method was used to determine the protein concentration of the crude extract of antimicrobial peptides from Apriona swainsoni larvae,and the filter paper method was used to determine its antibacterial activity. At the same time,its thermal stability,repeated freezethaw stability,acid-base stability,and other biochemical properties were explored. The results showed as follows:1)the protein concentration of the crude extract of antibacterial peptides from Apriona swainsonis larvae induced by Escherichia coli and Staphylococcus aureus was extremely significantly higher than that of the noninduced control group(P<0.01),and the protein concentration of the antibacterial peptide crude extract produced at 24 h of induction was the highest. 2)The crude extract of antibacterial peptides from Apriona swainsonis larvae had inhibitory effects on Staphylococcus aureus and B. subtilis,but there were no inhibitory effects on Escherichia coli and Pseudomonas aeruginosa. 3)The measurement results of biochemical properties showed that the crude extract of antibacterial peptides from Apriona swainsonis larvae had good thermal stability and acid-base stability and has little effect on its antibacterial activity after repeated freezing and thawing. In conclusion,mixing Escherichia coli and Staphylococcus aureus with equal volume can produce antimicrobial peptides with high concentration and strong activity,and their biochemical properties are relatively stab

关 键 词：斗米虫 抗菌肽 抑菌活性 生化特性 昆虫蛋白质资源 

分 类 号：Q819[生物学—生物工程]