Skip to main content
SpringerLink
Log in

An Enhanced Process for the Production of a Highly Purified Extracellular Lipase in the Non-conventional Yeast Yarrowia lipolytica

  • Published:
Applied Biochemistry and Biotechnology Aims and scope Submit manuscript

Abstract

Yarrowia lipolytica LgX64.81 is a non-genetically modified mutant that was previously identified as a promising microorganism for extracellular lipase production. In this work, the development of a fed-batch process for the production of this enzyme in this strain was described. A lipolytic activity of 2,145 U/mL was obtained after 32 h of batch culture in a defined medium supplemented with 10 g/L of tryptone, an enhancer of lipase expression. To maximize the volumetric productivity, two different fed-batch strategies had been investigated. In comparison to batch process, the intermittent fed-batch strategy had not improved the volumetric lipase productivity. In contrast, the stepwise feeding strategy combined with uncoupled cell growth and lipase production phases resulted in a 2-fold increase in the volumetric lipase productivity, namely, the lipase activity reached 10,000 U/mL after 80 h of culture. Furthermore, this lipase was purified to homogeneity by anion exchange chromatography on MonoQ resin followed by gel filtration on Sephacryl S-100. This process resulted in an overall yield of 72% and a 3.5-fold increase of the specific lipase activity. The developed process offers a great potential for an economic production of Lip2 at large scale in Y. lipolytica LgX64.81.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  1. Jaeger, K. E., & Eggert, T. (2002). Current Opinion in Biotechnology, 13, 390–397. doi:10.1016/S0958-1669(02)00341-5.

    Article  CAS  Google Scholar 

  2. Vakhlu, J., & Kour, A. (2006). Electronic Journal of Biotechnology, 9, 69–85. doi:10.2225/vol9-issue1-fulltext-9.

    Article  CAS  Google Scholar 

  3. Gombert, A. K., Pinto, A. L., Castilho, L. R., & Freire, D. M. G. (1999). Process Biochemistry, 35, 85–90. doi:10.1016/S0032-9592(99)00036-9.

    Article  CAS  Google Scholar 

  4. Reez, M. T. (2002). Current Opinion in Biotechnology, 6, 145–150. doi:10.1016/S1367-5931(02)00297-1.

    Article  Google Scholar 

  5. Vulfson, E. (1994). Industrial applications of lipases. In P. Wooley & S. B. Petersen (Eds.), Lipases (pp. 271–288). Cambridge: Cambridge University Press.

    Google Scholar 

  6. Zentler-Munro, P., Asssoufi, A., Balasubramanian, K., Cornell, D., Benoliel, D., Northfield, T., et al. (1992). Pancreas, 7, 311–319. doi:10.1097/00006676-199205000-00007.

    Article  CAS  Google Scholar 

  7. Vellard, M. (2003). Current Opinion in Biotechnology, 14, 444–450. doi:10.1016/S0958-1669(03)00092-2.

    Article  CAS  Google Scholar 

  8. Barth, G., & Gaillardin, C. (1997). FEMS Microbiology Reviews, 19, 219–237. doi:10.1111/j.1574-6976.1997.tb00299.x.

    Article  CAS  Google Scholar 

  9. Pignède, G., Wang, H., Fudalej, F., Gaillardin, C., Seman, M., & Nicaud, J. M. (2000). Journal of Bacteriology, 182, 2802–2810. doi:10.1128/JB.182.10.2802-2810.2000.

    Article  Google Scholar 

  10. Fickers, P., Fudalej, F., Le Dall, M. T., Casaregola, S., Gaillardin, C., Thonart, P., et al. (2005). Fungal Genetics and Biology, 42, 264–274. doi:10.1016/j.fgb.2004.12.003.

    Article  CAS  Google Scholar 

  11. Destain, J., Roblain, D., & Thonart, P. (1997). Biotechnology Letters, 19(2), 105–107. doi:10.1023/A:1018339709368.

    Article  CAS  Google Scholar 

  12. Nicaud, J. M., Madzak, C., Broek, P., Gysler, C., Duboc, P., Niederberger, P., et al. (2002). FEMS Yeast Research, 2, 371–379.

    CAS  Google Scholar 

  13. Fickers, P., Fudalej, F., Nicaud, J. M., Destain, J., & Thonart, P. (2005). Journal of Biotechnology, 115, 379–386. doi:10.1016/j.jbiotec.2004.09.014.

    Article  CAS  Google Scholar 

  14. Novotný, C., Dolezalovă, L., Musil, P., & Novăk, M. (1988). Journal of Basic Microbiology, 28(4), 221–227. doi:10.1002/jobm.3620280403.

    Article  Google Scholar 

  15. Fickers, P., Nicaud, J. M., Gaillardin, C., Destain, J., & Thonart, P. (2004). Journal of Applied Microbiology, 96, 742–749. doi:10.1111/j.1365-2672.2004.02190.x.

    Article  CAS  Google Scholar 

  16. Pereira-Meirelles, F. V., Rocha-Leăo, M. H. M., & Sant’Anna, G. L. Jr. (2000). Biotechnology Letters, 23, 71–75. doi:10.1023/A:1005672731818.

    Article  Google Scholar 

  17. Dominguez, A., Deive, F. J., Sanroman, M. A., & Longo, M. A. (2003). Journal of Chemical Technology and Biotechnology (Oxford, Oxfordshire), 78, 1166–1170. doi:10.1002/jctb.922.

    Article  CAS  Google Scholar 

  18. Fickers, P., Nicaud, J. M., Destain, J., & Thonart, P. (2005). Current Microbiology, 50, 133–137.

    CAS  Google Scholar 

  19. Fickers, P., Ongena, M., Destain, J., Weekers, F., & Thonart, P. (2006). Enzyme and Microbial Technology, 36, 756–759. doi:10.1016/j.enzmictec.2005.08.005.

    Article  Google Scholar 

  20. Destain, J., Fickers, P., Weekers, F., Moreau, B., & Thonart, P. (2005). Applied Biochemistry and Biotechnology, 121–124, 269–277. doi:10.1385/ABAB:121:1-3:0269.

    Article  Google Scholar 

  21. Huber, H., Weigl, G., & Buchinger, W. (2005). WO2005097990.

  22. Gordillo, M. A., Sanz, A., Sảnchez, A., Valero, F., Montesinos, J. L., Lafuente, J., et al. (1998). Biotechnology Bioengineering, 60(2), 156–168. doi:10.1002/(SICI)1097-0290(19981020)60:2<156::AID-BIT3>3.0.CO;2-M.

    Article  CAS  Google Scholar 

  23. Kortz, D. J., Rinas, U., Hellmuth, K., Sanders, E. A., & Deckwer, W. D. (1995). Journal of Biotechnology, 39, 59–65. doi:10.1016/0168-1656(94)00143-Z.

    Article  Google Scholar 

  24. Frings, C., & Dunn, R. (1970). American Journal of Clinical Pathology, 53, 89–91.

    CAS  Google Scholar 

  25. Laemmli, U. K. (1970). Nature, 227, 680–685. doi:10.1038/227680a0.

    Article  CAS  Google Scholar 

  26. Turki, S., Ben Kraeim, I., Weekers, F., Thonart, P., & Kallel, H. (2009). Bioresource Technology, 100, 2724–2731. doi:10.1016/j.biortech.2008.12.053.

    Article  CAS  Google Scholar 

  27. Saxena, R. K., Sheoran, A., Giri, B., & Davidson, W. S. (2003). Journal of Microbiological Methods, 52, 1–18. doi:10.1016/S0167-7012(02)00161-6.

    Article  CAS  Google Scholar 

  28. Hahn-Hägerdal, B., Karhumaa, K., Larsson, C. U., Gorwa-Grauslund, M., Görgens, J., & Zyl, W. H. (2005). Microbial Cell Factories, 4, 31.

    Article  Google Scholar 

  29. Zhang, J., & Greasham, R. (1999). Applied Microbial Biotechnology, 51(4), 407–421. doi:10.1007/s002530051411.

    Article  CAS  Google Scholar 

  30. Destain, J. (1998), Ph.D. thesis, Faculté Universitaire des Sciences Agronomiques de Gembloux. Belgium, Gembloux.

  31. Freire, D. M., Teles, E. M. F., Bon, E. P. S., & Sant’Anna, G. L. Jr. (1997). Applied Biochemistry and Biotechnology, 63–65, 409–421. doi:10.1007/BF02920442.

    Article  Google Scholar 

  32. Montesinos, J. L., Gordillo, M. A., Valero, F., Lafuente, J., & Solà, C. (1997). Journal of Biotechnology, 52, 207–218. doi:10.1016/S0168-1656(96)01646-X.

    Article  CAS  Google Scholar 

  33. Gordillo, M. A., Montesinos, J. L., Casas, C., Valero, F., Lafuente, J., & Solà, C. (1998). Chemistry and Physics of Lipids, 93, 131–142. doi:10.1016/S0009-3084(98)00037-1.

    Article  CAS  Google Scholar 

  34. Aguedo, M., Waché, Y., Mazoyer, V., Sequeira-Le Grand, A., & Belin, J. M. (2003). Journal of Agricultural and Food Chemistry, 51, 3007–3011. doi:10.1021/jf020901m.

    Article  CAS  Google Scholar 

  35. Fickers, P., Destain, J., & Thonart, P. (2008). Journal of Basic Microbiology, 48, 1–4. doi:10.1002/jobm.200890001.

    Article  Google Scholar 

  36. Fickers, P., Nicaud, J. M., Destain, J., & Thonart, P. (2003). Applied Microbiology and Biotechnology, 63, 136–142. doi:10.1007/s00253-003-1342-3.

    Article  CAS  Google Scholar 

  37. Aloulou, A., Rodriguez, J. A., Puccinelli, D., Mouz, N., Leclaire, J., Leblond, Y., et al. (2007). Biochimica et Biophysica Acta, 1771, 228–237.

    CAS  Google Scholar 

  38. Yu, M., Lange, S., Richter, S., Tan, T., & Schmid, R. D. (2007). Protein Expression and Purification, 53(2), 63–255. doi:10.1016/j.pep.2006.10.018.

    Article  Google Scholar 

  39. Gerngross, T. U. (2004). Nature Biotechnology, 22(11), 1409–1414. doi:10.1038/nbt1028.

    Article  CAS  Google Scholar 

Download references

Acknowledgment

The authors are grateful to the “Commissariat Général aux Relations Internationales de la Communauté Française de Belgique” for financial support.

Author information

Authors and Affiliations

  1. Unité de Fermentation, Institut Pasteur de Tunis, 13, place Pasteur, BP 74, 1002, Tunis Belvédère, Tunisia

    Saoussen Turki, Atef Ayed, Néjib Chalghoumi & Héla Kallel

  2. Artechno S.A., Rue François Bovesse, 1. 5030, Gembloux, Belgium

    Frederic Weekers

  3. Centre Wallon de Biologie Industrielle, Unité de Technologie Microbienne, Université de Liège, Bd du Rectorat Bat. 40, 4000, Liège, Belgium

    Philippe Thonart

Authors
  1. Saoussen Turki
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Atef Ayed
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Néjib Chalghoumi
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Frederic Weekers
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Philippe Thonart
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Héla Kallel
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Héla Kallel.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Turki, S., Ayed, A., Chalghoumi, N. et al. An Enhanced Process for the Production of a Highly Purified Extracellular Lipase in the Non-conventional Yeast Yarrowia lipolytica . Appl Biochem Biotechnol 160, 1371–1385 (2010). https://doi.org/10.1007/s12010-009-8599-7

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12010-009-8599-7

Keywords

Search

Navigation