1992年发现了一种由海洋细菌Alcanivoraxborkumensis合成的新型生物表面活性剂。这种细菌在受石油泄漏影响的海洋环境中含量丰富,它有助于降解烷烃,在这样的条件下,产生甘氨酸-葡萄糖脂生物表面活性剂。生物表面活性剂增强细菌对油滴的附着并促进碳氢化合物的吸收。由于其预期的有用特性,人们对这种生物表面活性剂的生物技术生产感兴趣。为了分析支持这一努力,开发了一种将反相高效液相色谱(HPLC)与高分辨率质谱(HRMS)相结合的方法,允许分离和鉴定甘氨酸-葡萄糖脂同源物。准确的质量,保留时间,和特征性的碎片模式用于物种分配。此外,采用带电气溶胶检测(CAD)来实现无真实标准的绝对定量。该方法用于研究使用不同碳源的A.borkumensisSK2的甘氨酸-葡萄糖脂质生产。质谱使我们能够鉴定掺入的3-羟基-链烷酸中具有不同链长(C6-C12)和不饱和度(0-1个双键)的同源物,一些以前未知的。使用CAD的定量显示,当用十六烷生长时,滴度约为丙酮酸盐的两倍(49mg/L对22mg/L)。两种碳源的主要同源物是glc-40:0-gly,丙酮酸占64%,十六烷作为唯一碳源占85%。有了这里展示的分析服,可以识别复杂和变化的糖脂,characterized,量化,如本文示例性示出的A.borkumensis的有趣的甘氨酸-葡萄糖脂。
A novel biosurfactant was discovered to be synthesized by the marine bacterium Alcanivorax borkumensis in 1992. This bacterium is abundant in marine environments affected by oil spills, where it helps to degrade alkanes and, under such conditions, produces a glycine-glucolipid biosurfactant. The biosurfactant enhances the bacterium\'s attachment to oil droplets and facilitates the uptake of hydrocarbons. Due to its useful properties expected, there is interest in the biotechnological production of this biosurfactant. To support this effort analytically, a method combining reversed-phase high-performance liquid chromatography (HPLC) with high-resolution mass spectrometry (HRMS) was developed, allowing the separation and identification of glycine-glucolipid congeners. Accurate mass, retention time, and characteristic fragmentation pattern were utilized for species assignment. In addition, charged-aerosol detection (CAD) was employed to enable absolute quantification without authentic standards. The methodology was used to investigate the glycine-glucolipid production by A. borkumensis SK2 using different carbon sources. Mass spectrometry allowed us to identify congeners with varying chain lengths (C6-C12) and degrees of unsaturation (0-1 double bonds) in the incorporated 3-hydroxy-alkanoic acids, some previously unknown. Quantification using CAD revealed that the titer was approximately twice as high when grown with hexadecane as with pyruvate (49 mg/L versus 22 mg/L). The main congener for both carbon sources was glc-40:0-gly, accounting for 64% with pyruvate and 85% with hexadecane as sole carbon source. With the here presented analytical suit, complex and varying glycolipids can be identified, characterized, and quantified, as here exemplarily shown for the interesting glycine-glucolipid of A. borkumensis.