枯草芽孢杆菌HS-A38产抗菌脂肽Bacillomycin D的组分分析及鉴定

对一株枯草芽孢杆菌HS-A38产生的脂肽类物质进行分离鉴定及抑菌活性研究。通过酸沉淀分离和有机溶剂抽提的方法,从枯草芽孢杆菌HS-A38发酵液中得到脂肽粗提物LP,产率为1.956 g/L。利用薄层色谱和茚三酮染色法确定该脂肽粗提物中存在四个组分,分别为LP1、LP2、LP3和LP4;抑菌活性检测显示,组分LP3对两株海洋致病菌副溶血性弧菌和铜绿假单胞杆菌的活性较高。组分LP3经硅胶柱层析纯化分离后,应用反相高效液相色谱(RP-HPLC)和基质辅助激光解析电离飞行时间质谱(MALDI-TOF-MS)对该组分做进一步纯化和鉴定。分析表明,LP3样品在保留时间20 min~28 min产生单峰团LP3-1,其纯度为85.24%;经MALDI-TOF-MS分析和数据比对,组分LP3-1中的主要成分为杆菌霉素Bacillomycin D。

Analysis and identification of lipopeptide Bacillomycin D produced by Bacillus subtilis HS-A38

In this study the lipopeptide compounds produced by the strain Bacillus subtilis HS-A38 was isolated and identified,and their antibacterial activities were analyzed.The lipopeptide crude extract LP was isolated by using the acid precipitation and organic solvent extraction from the strain HS-A38 fermentation culture,and its yield was 1.956 g/L.Four components were identified by the ninhydrin color printing and thin layer chromatography,named as LP1,LP2,LP3 and LP4.The results of antibacterial activity test indicated that LP3 had higher activity against two marine pathogenic bacteria of Vibrio parahaemolyticus and Pseudomonas aeruginosa.The LP3 was separated by the Silica Gel Column Chromatography,and then further purified and identified by using RP-HPLC and MALDI-TOF-MS.According to the RPHPLC analysis,the peak group of LP3-1 was found within 20 min ～ 28 min.Based on the mass spectra analysis,the main composition of the LP3-1 was Bacillomycin D.