摘要
目的:比较两种提取液对黄粉虫抗菌肽的提取效率及其抑菌活性。方法:用大肠杆菌DH-5α菌液(1×108/ml)腹部注射黄粉虫5龄幼虫,72 h后分别以两种提取液破碎全虫提取抗菌肽。提取液A为5%乙酸,提取液B为0.05mol/L乙酸铵缓冲液,pH 5.0、35μg/ml苯甲基磺酰氟、2‰巯基乙醇、0.2 mg/L EDTA、1 mg/ml苯基硫脲;两种提取液提取后分别测定未诱导和诱导抗菌肽浓度后,4样品均配制成80μg/ml的抗菌肽溶液,对大肠杆菌DH-5α进行体外抑菌实验。结果:两种提取液提取到的未诱导组间抗菌肽浓度分别为(114.48±10.95)μg/ml和(124.85±9.31)μg/ml,组间无统计学意义(P>0.05),诱导组间抗菌肽浓度分别为(436.98±11.62)μg/ml和(612.73±9.49)μg/ml,差异具有统计学意义(P<0.01);将上述两组抗菌肽溶液配制成相同浓度(80μg/ml)分别对大肠杆菌DH-5α体外抑菌试验,结果显示:未诱导组间抑菌作用具有统计学意义(P<0.05),诱导组间的抑菌作用亦具有统计学意义(P<0.01)。结论:提取液B提取到的抗菌肽不管在抗菌肽提取效率上,还是在抑菌效果方面均优于提取液A,从而为抗菌肽提取工艺的优化提供参考。
Objective：To compare the extracting efficiency of antimicrobial peptides from Tenebrio molitor using two different extracting solutions and detect their antibacterial activities.Methods：E.coli DH-5α（1× 10^8/ml） was abdominally injected into the Tenebrio molitor at its fifth instar larvae.After 72 h,two diverse solutions were applied to extracting the antimicrobial peptides.Extracting buffer A was 5% acetic acid（V/V）,and buffer B included ammonium acetate at pH 5.0 in concentration of 0.05 mol/L,35 μg/ml phenylmethylsulfonyl fluoride,2‰ mercaptoethanol,0.2 mg/L EDTA and 1 mg/ml phenyl thiourea.The concentrations of antimicrobial peptides（including un-induced and induced groups） were measured after extraction using two the different solutions.The four samples were separately used to prepare the antibacterial peptide solution to a concentration of 80 μg/ml,and in vitro antibacterial assay（DH-5α as sample） was performed to determined the extracting efficiencies by the protocols described above.Results：The concentration of antibacterial peptides from un-induced groups was（114.48 ± 10.95） μg/ml and（124.85 ± 9.31） μg/ml,respectively,which suggested no statistical difference（P〉0.05）,whereas the concentration in the induced groups was（436.98 ± 11.62） μg/ml and（612.73 ± 9.49） μg/ml,respectively,with significant statistical difference（P〈0.01）.The test of in vitro antibacterial effect on E.coli DH-5α showed the similar outcomes for both un-induced and induced groups and also had statistical difference within the group（P〈0.05,P〈0.01,respectively）.Conclusion：Although both buffers had led to similar efficiencies,yet buffer B looks superior to buffer A,to a certain extent,regarding the efficiency and antibacterial effects.
出处
《皖南医学院学报》
CAS
2012年第4期263-265,共3页
Journal of Wannan Medical College
基金
安徽高校省级自然科学研究项目(KJ2012Z426)
关键词
提取液
黄粉虫
抗菌肽
extracting solution
Tenebrio molitor
antibacterial peptide
