贝莱斯芽孢杆菌Ndl5发酵液中脂肽生物表面活性剂的纯化与鉴定

Purification and identification of lipopeptide biosurfactant in the fermentation broth of Bacillus velezensis Ndl5

试验利用6mol/L盐酸使贝莱斯芽孢杆菌Ndl5发酵液中的脂肽表面活性剂沉淀,沉淀经甲醇提取获得脂肽粗提物,经过UniElut C18 HC固相萃取初步分离,最后利用反相HPLC半制备纯化,收集馏分S3-L1和S3-L3。采用超高效液相串联四极杆飞行时间质谱(UHPLC-Q-TOF-MS)在正离子模式下测得S3-L1质荷比(m/z)为1022.6763,S3-L3质荷比(m/z)为1036.6919,判断S3-L1结构C_(14)-Glu^(1)-Leu^(2)-Leu^(3)-Val^(4)-Asp^(5)-Leu^(6)-Leu^(7),S3-L3结构为C_(15)-Glu^(1)-Leu^(2)-Leu^(3)-Val^(4)-Asp^(5)-Leu^(6)-Ile^(7)。

The experiment used 6 mol/L hydrochloric acid to precipitate the lipopeptide surfactant in the Bacillus velezensis Ndl5 fermentation broth,the precipitate was then extracted with methanol to obtain the crude lipopeptide extract,which was preliminarily separated by UniElut C18 HC solid phase extraction,and finally reversed phase HPLC semi-preparative purification,collect fractions S3-L1 and S3-L3.Further using ultra-high performance liquid phase tandem quadrupole time-of-flight mass spectrometry(UHPLC-Q-TOF-MS)in the positive ion mode,the mass-to-charge ratio(m/z)of S3-L1 was 1022.6763,and the mass-to-charge ratio of S3-L3(m/z)was 1036.6919,the S3-L1 structure is judged to be C_(14)-Glu^(1)-Leu^(2)-Leu^(3)-Val^(4)-Asp^(5)-Leu^(6)-Leu^(7),and the S3-L3 structure was C_(15)-Glu^(1)-Leu^(2)-Leu^(3)-Val^(4)-Asp^(5)-Leu^(6)-Ile^(7).