摘要
长期大量使用杀菌剂不但会导致病原微生物产生耐药性,同时也存在着严重的环境和健康风险,因此开发不易产生耐药性的新型抗菌物质非常重要。本实验采用硫酸铵沉淀、AB-8大孔吸附树脂、Sephadex G-100凝胶和半制备型反相高效液相色谱对短小芽孢杆菌(Bacillus pumilus)HN-10发酵液进行逐级分离纯化,以粉红单端孢(Trichothecium roseum)为指示菌,测其抑菌活性,并通过基质辅助激光解析串联飞行时间串联质谱对纯化后的活性物质进行氨基酸序列分析。结果表明,经纯化后得到一种抗真菌肽P-1,其氨基酸序列为G-G-S-G-G-G-S-S-G-G-S-I-G-G-R,分子质量为1 149.14 Da。经抑菌实验可知,抗真菌肽P-1在最小抑菌浓度为1μg/mL时可抑制粉红单端孢生长。经ProtParam软件分析得到P-1理论等电点为9.75,带有1个正电荷数。在NCBI数据库和APD数据库检索,与之序列相似性最高的仅为45%,因此判断这是一种新型抗真菌肽。抗真菌肽P-1抑菌效果良好,可为开发新型生防制剂提供理论依据。
Chronic heavy use of bactericides could not only induce antibiotic resistance,but also could cause serious environmental and health risks.Thus,it is very important to develop new bactericides that kill pathogens without detectable resistance.In this study,a novel antimicrobial peptide produced by Bacillus pumilus HN-10 was purified consecutively by ammonium sulfate precipitation,macroporous resin AB-8 chromatography,Sephadex G-100 chromatography,and reversedphase high performance liquid chromatography(RP-HPLC).The antimicrobial activity of the peptide was tested against Trichothecium roseum.The amino acid sequence was analyzed by matrix assisted laser desorption ionization time of flight mass spectrometry(MALDI-TOF-MS/MS).The results showed that this peptide was designated as P-1 and its amino acid sequence was G-G-S-G-G-G-S-S-G-G-S-I-G-G-R with a molecular mass of 1 149.14 Da.P-1 exhibited strong antimicrobial activity against T.roseum with a minimum inhibitory concentration of 1μg/mL.Analysis with the ProtParam software indicated that the theoretical isoelectric point of P-1 was 9.75,with one positive charge.P-1 was proved to be a novel peptide by NCBI database and APD database search with the highest similarity of only 45%.
作者
严海娇
贠建民
白杰
郭娟
邓展瑞
李多佳
YAN Haijiao;YUN Jianmin;BAI Jie;GUO Juan;DENG Zhanrui;LI Duojia(College of Food Science and Engineering,Gansu Agricultural University,Lanzhou 730070,China)
出处
《食品科学》
EI
CAS
CSCD
北大核心
2018年第22期123-128,共6页
Food Science
基金
国家自然科学基金地区科学基金项目(31360405)
关键词
抗菌肽
短小芽孢杆菌HN-10
分离纯化
粉红单端孢
antimicrobial peptide
Bacillus pumilus HN-10
characterization and purification
Trichothecium roseum
