地衣芽孢杆菌抗菌肽的纯化及抗菌特性分析

Purification of Antimicrobial Peptide from Bacillus licheniformis LY12 and Analysis of Its Antibacterial Properties

以研究地衣芽孢杆菌(Bacillus licheniformis)LY12所产抗菌肽特性,从发酵液中分离纯化该抗菌肽。通过超滤、离子交换层析、分子筛凝胶过滤层析、反向高效液相色谱等方法纯化该抗菌肽。结果显示:LC-ESI-MS质谱测得抗菌肽分子量为2750.785 Da,同Tricine SDS-PAGE分析结果基本一致。体外抗菌试验结果显示,抗菌肽LYF12能够抑制丝状真菌、细菌的生长。对枯草芽孢杆菌(Bacillus subtilis)、大肠杆菌(Escherichia coli)、铜绿假单胞菌(Pseudomonas aeruginosa)、金黄色葡萄球菌(Staphylococcus aureus)、藤黄微球菌(Micrococcus aureus)、蜡状芽孢杆菌(Bacillus cereus)、副溶血性弧菌(Vibrio parachaemolyticus)的最低抑菌浓度分别为9.8、19.8、20.7、7.2、9.6、10.2、15.7μg/mL。从地衣芽孢杆菌LY12分离的抗菌肽显示出对革兰氏阴性细菌、革兰氏阳性细菌及真菌具有较好的抗菌效果,故在食品防腐中具有潜在的应用价值。

To explore the characterization of a novel antimicrobial peptide from from Bacillus licheniformis LY12,the antimicrobial peptide was isolated. The novel antimicrobial peptide designated LYF12,was purified,with a procedure involving ultrafiltration,ion exchange chromatography（DEAE-Sepharose FF）,gel filtration（Sephadex G-15）,and reverse phase HPLCs on C8column and C18column. LC-ESI-MS indicated the molecular mass of the purified peptide was 2750.785 Da,which was in good agreement with the result of Tricine SDS-PAGE..In vitro bioassays showed that LYF12 inhibits the growth of a variety of microbes,including filamentous fungi and bacteria. The minimum inhibitory concentrations（MIC） against Bacillus subtilis,Escherichia coli,Pseudomonas aeruginosa,Staphylococcus aureus,Micrococcus aureus,Bacillus cereus,Vibrio parachaemolyticus were 9.8,19.8,20.7,7.2,9.6,10.2,15.7 μg/mL,respectively. Because the purified antimicrobial peptide exhibited antibacterial activity against Gram-positive and Gram-negativebacteria as well as fungi,the antimicrobial peptide from Bacillus licheniformis LY12 may have potential application in food industry.