Citation: 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. doi: 10.3724/SP.J.1123.2013.10026 shu

Purification and characterization of the biosurfactant rhamnolipid

1.
1. College of Environmental Science and Engineering, Hunan University, Changsha 410082, China;

Corresponding author: ZHONG Hua, ZENG Guangming,
Received Date: 29 October 2013
Available Online: 20 December 2013
Fund Project: 国家自然科学基金项目(51378190,50908081,51039001,51378192,51308200). (51378190,50908081,51039001,51378192,51308200)

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.
- acid precipitation,
- column chromatography,
- high performance liquid chromatography-mass spectrometry (HPLC-MS),
- rhamnolipid,
- Pseudomonas aeruginosa,
- biosurfactant
1. [1]
  [1] Makkar R S, Cameotra S S, Banat I M. AMB Express, 2011, 1(5): 18
2. [2]
  [2] Liu Z F, Zeng G M, Zhong H, et al. World J Microb Biot, 2012, 28: 175
3. [3]
  [3] Liu X L, Zeng G M, Tang L, et al. Process Biochem, 2008, 43: 1300
4. [4]
  [4] Liang Y S, Yuan X Z, Zeng G M, et al. Biodegradation, 2010, 21: 615
5. [5]
  [5] Banat I M, Franzetti A, Gandolfi I, et al. Appl Microbiol Biot, 2010, 87: 427
6. [6]
  [6] Liu Z F, Zeng G M, Wang J, et al. Process Biochem, 2010, 45: 805
7. [7]
  [7] Chayabutra C, Wu J, Ju L K. Biotechnol Bioeng, 2001, 72 (1): 25
8. [8]
  [8] Sarachat T, Pornsunthorntawee O, Chavadej S, et al. Bioresource Technol, 2010, 101: 324
9. [9]
  [9] Wang W, Zeng G M, Huang G H, et al. Acta Scientiae Circumstantiae (王伟, 曾光明, 黄国和, 等. 环境科学学报), 2005, 25(7): 965
10. [10]
  [10] Nitschke M, Costa S G V A O, Contiero J. Appl Microbiol Biot, 2010, 160: 2066
11. [11]
  [11] Ma M Y, Shi Z, Liu Y S. Chinese Journal of Environmental Engineering (马满英, 施周, 刘有势. 环境工程学报), 2008, 2(1): 83
12. [12]
  [12] Kuyukina M S, Ivshina I B, Philp J C, et al. J Microbiol Meth, 2001, 46: 149
13. [13]
  [13] Schenk T, Schuphan I, Schmidt B. J Chromatogr A, 1995, 693: 7
14. [14]
  [14] Davis D A, Lynch H C, Varley J. Enzyme Microb Tech, 2001, 28: 346
15. [15]
  [15] Hubert J, Plé K, Hamzaoui M, et al. C R Chimie, 2012, 15: 18
16. [16]
  [16] Witek-Krowiak A, Witek J, Gruszczyńska A, et al. World J Microb Biot, 2011, 27: 1961
17. [17]
  [17] Long X W, Meng Q, Sha R Y, et al. J Membrane Sci, 2012, 409: 105
18. [18]
  [18] Mukherjee S, Das P, Sen R. Trends Biotechnol, 2006, 24(11): 509
19. [19]
  [19] Déziel E, Lépine F, Dennie D, et al. Biochim Biophys Acta, 1999, 1440: 244
20. [20]
  [20] Abalos A, Pinazo A, Infante M R, et al. Langmuir, 2001, 17: 1367
21. [21]
  [21] Singh N, Pemmaraju S C, Pruthi P A, et al. Appl Microbiol Biot, 2013, 169: 2374
22. [22]
  [22] Choi M H, Xu J, Gutierrez M, et al. J Biotechnol, 2011, 151(1): 30
23. [23]
  [23] Ma H N, Hua Y J, Tu C Y, et al. Chinese Journal of Chromatography (马海宁, 华玉娟, 屠春燕, 等. 色谱), 2012, 30(3): 304
24. [24]
  [24] Abdel-Mawgoud A M, Hausmann R, Lépine F, et al. Biosurfactants: from Genes to Applications. Berlin: Springer Verlag Heidelberg, 2011: 21
25. [25]
  [25] Heyd M, Kohnert A, Tan T H, et al. Anal Bioanal Chem, 2008, 391: 1579
26. [26]
  [26] Nitschke M, Costa S G V A O, Haddad R, et al. Biotechnol Progr, 2005, 21: 1562
27. [27]
  [27] Mata-Sandoval J C, Karns J, Torrents A. J Chromatogr A, 1999, 864: 211
28. [28]
  [28] Zhong H, Zeng G M, Liu J X, et al. Appl Microbiol Biot, 2008, 79: 671
29. [29]
  [29] Zhong H, Zeng G M, Yuan X Z, et al. Appl Microbiol Biot, 2007, 77: 447
30. [30]
  [30] Fu H Y, Zeng G M, Yuan X Z, et al. Journal of Biology (傅海燕, 曾光明, 袁兴中, 等. 生物学杂志), 2003, 20(6): 1
31. [31]
  [31] Arino S, Marchal R, Vandecasteele J P. Appl Microbiol Biot, 1996, 45: 162
32. [32]
  [32] Kennedy J H, Wiseman J M. Rapid Commun Mass Spectrom, 2010, 24: 1305
33. [33]
  [33] Noordman W H, Brusseau M L, Janssen D B. Environ Sci Technol, 2000, 34: 832
34. [34]
  [34] Ishigami Y, Gama Y, Nagahora H, et al. Chem Lett, 1987: 763
35. [35]
  [35] Soberón-Chávez G, Lépine F, Déziel E. Appl Microbiol Biot, 2005, 68: 718
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