Liu Yang, Zhong Hua, Liu Zhifeng, Jiang Yongbing, Tan Fei, Zeng Guangming, Lai Mingyong, He Yibin
Se Pu. 2014 Mar;32(3):248-55. doi: 10.3724/sp.j.1123.2013.10026.
Biosurfactant rhamnolipid is a metabolic intermediate produced by microorganisms under a certain condition. There are the polar hydrophilic group and the non-polar hydrophobic group in rhamnolipid molecule which always exhibits high surface or interfacial activity. A reliable separation and purification method as well as component identification technique is essential for success of production process. The rhamnolipid was produced by aerobic fermentation using Pseudomonas aeruginosa CCTCC AB93066 in this study. It was separated from the culture by acid precipitation and purified by column chromatography until two groups of monorhamnolipid and dirhamnolipid were obtained. High performance liquid chromatography with mass spectrometry (HPLC-MS) examination showed that either the monorhamnolipid or the dirhamnolipid contained three major species. They were RhaC10C10, RhaC10C12-H2, RhaC10C12 for monorhamnolipid and Rha2C10 C10, Rha2C10 C12-H2, Rha2 C10 C12 for dirhamnolipid. The results of the study suggested that Pseudomonas aeruginosa CCTCC AB93066 is a good strain for rhamnolipid production. Acid precipitation-column chromatography technique is good for purification of rhamnolipid. Meanwhile, HPLC-MS is a reliable method for identifying components of rhamnolipid with high sensitivity and accuracy.
生物表面活性剂鼠李糖脂是微生物在一定条件下产生的代谢中间体。鼠李糖脂分子中存在极性亲水基团和非极性疏水基团,其总是表现出高表面活性或界面活性。一种可靠的分离纯化方法以及成分鉴定技术对于生产过程的成功至关重要。本研究中,鼠李糖脂由铜绿假单胞菌CCTCC AB93066通过好氧发酵产生。通过酸沉淀从培养物中分离出来,并通过柱色谱法纯化,直到获得两组单鼠李糖脂和双鼠李糖脂。高效液相色谱-质谱联用(HPLC-MS)检测表明,单鼠李糖脂和双鼠李糖脂均包含三种主要成分。单鼠李糖脂的三种主要成分是RhaC10C10、RhaC10C12-H2、RhaC10C12,双鼠李糖脂的三种主要成分是Rha2C10 C10、Rha2C10 C12-H2、Rha2 C10 C12。研究结果表明,铜绿假单胞菌CCTCC AB93066是生产鼠李糖脂的优良菌株。酸沉淀-柱色谱技术有利于鼠李糖脂的纯化。同时,HPLC-MS是一种可靠的鼠李糖脂成分鉴定方法,具有高灵敏度和准确性。