生物表面活性剂鼠李糖脂的纯化与表征

doi:10.3724/SP.J.1123.2013.10026

色谱 ›› 2014, Vol. 32 ›› Issue (3): 248-255.DOI: 10.3724/SP.J.1123.2013.10026

生物表面活性剂鼠李糖脂的纯化与表征

刘洋, 钟华, 刘智峰, 蒋勇兵, 谈菲, 曾光明, 赖明勇, 何益斌

1. 湖南大学环境科学与工程学院, 湖南长沙 410082;
2. 环境生物学与控制教育部重点实验室(湖南大学), 湖南长沙 410082

收稿日期:2013-10-29 修回日期:2013-12-20 出版日期:2014-03-08 发布日期:2014-03-04
通讯作者: 曾光明，钟华
基金资助:
国家自然科学基金项目（51378190，50908081，51039001，51378192，51308200）.

Purification and characterization of the biosurfactant rhamnolipid

LIU Yang, ZHONG Hua, LIU Zhifeng, JIANG Yongbing, TAN Fei, ZENG Guangming, LAI Mingyong, HE Yibin

1. College of Environmental Science and Engineering, Hunan University, Changsha 410082, China;
2. Key Laboratory of Environmental Biology and Pollution Control (Hunan University), Ministry of Education, Changsha 410082, China

Received:2013-10-29 Revised:2013-12-20 Online:2014-03-08 Published:2014-03-04

摘要/Abstract

摘要：

生物表面活性剂鼠李糖脂是微生物在一定条件下产生的次级代谢产物，其分子具有极性亲水基团和非极性亲油基团结构，通常表现出很高的表面活性和界面优先分配能力。可靠的分离提纯方法和成分鉴定手段是鼠李糖脂生产工艺成功的重要保证。实验通过好氧发酵培养铜绿假单胞菌CCTCC AB93066、酸沉降分离得到鼠李糖脂后，利用柱色谱提纯技术得到纯化的鼠李糖脂的单糖脂和二糖脂，最后采用高效液相色谱-质谱联用法进行成分鉴定。结果显示这两种鼠李糖脂均含有3种主要成分，其中单糖脂的主要成分为RhaC₁₀C₁₀、RhaC₁₀C₁₂-H₂、RhaC₁₀C₁₂，二糖脂的主要成分为Rha₂C₁₀C₁₀、Rha₂C₁₀C₁₂-H₂、Rha₂C₁₀C₁₂。该研究结果表明，铜绿假单胞菌CCTCC AB93066是一种良好的鼠李糖脂产生菌；酸沉降-柱色谱技术可以用于鼠李糖脂的深度提纯，且有较好的效果；而高效液相色谱-质谱联用技术对鼠李糖脂成分鉴定具有灵敏度高和准确性好等优点，是一种较为可靠的检测方法。

关键词: 高效液相色谱-质谱联用, 生物表面活性剂, 鼠李糖脂, 酸沉降, 铜绿假单胞菌, 柱色谱

Abstract:

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 Rha₂C₁₀C₁₀, Rha₂C₁₀C₁₂-H₂, Rha₂C₁₀C₁₂ 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.

Key words: Pseudomonas aeruginosa, acid precipitation, biosurfactant, column chromatography, high performance liquid chromatography-mass spectrometry (HPLC-MS), rhamnolipid

中图分类号:

O658

刘洋, 钟华, 刘智峰, 蒋勇兵, 谈菲, 曾光明, 赖明勇, 何益斌. 生物表面活性剂鼠李糖脂的纯化与表征[J]. 色谱, 2014, 32(3): 248-255.

LIU Yang, ZHONG Hua, LIU Zhifeng, JIANG Yongbing, TAN Fei, ZENG Guangming, LAI Mingyong, HE Yibin. Purification and characterization of the biosurfactant rhamnolipid[J]. Chinese Journal of Chromatography, 2014, 32(3): 248-255.

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Purification and characterization of the biosurfactant rhamnolipid

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