Applied Biochemistry and Biotechnology

, Volume 184, Issue 4, pp 1332–1346 | Cite as

Harnessing the Effect of pH on Lipid Production in Batch Cultures of Yarrowia lipolytica SKY7

  • Mathiazhakan Kuttiraja
  • Ayed Dhouha
  • Rajeshwar Dayal Tyagi


The objective of this research was to investigate the kinetics of lipid production by Yarrowia lipolytica SKY7 in the crude glycerol-supplemented media with and without the control of pH. Lipid and citric acid production were improved with the pH control condition. There was no significant difference observed in the biomass concentration with or without the pH control. In the pH-controlled experiments, the biomass and lipid concentration reached 18 and 7.78 g/L, (45.5% w/w), respectively, with lipid yield (Yp/s) of 0.179 g/g at 60 h of fermentation. The lipid production was directly correlated with growth and the process was defined as growth associated. After 60 h of fermentation, the lipid degradation was noticed in the pH-controlled reactor whereas it occurred after 84 h in the pH-uncontrolled reactor. Apart from lipid, citric acid was produced as the major extracellular product in both fermentations but the much lower concentration in uncontrolled pH. Based on the experimental results, it is evident that controlling the pH will enhance the lipid production by 15% compared to pH-uncontrolled fermentation.


Y. lipolytica Crude glycerol Biodiesel Microbial lipid C/N ratio Effect of pH 



The authors would like to acknowledge the Natural Sciences and Engineering Research Council of Canada (grant A4984, strategic grant 412994–11, Canada Research Chair) for financial support. We are grateful to the technical staffs of INRS-ETE for their timely help to analyze the samples on GC-FID.

Compliance with Ethical Standards

Conflict of Interest

All authors have read and agreed with the contents of the manuscript. The authors indicate no potential conflicts of interest.


  1. 1.
    Ayoub, M., & Abdullah, A. Z. (2012). Critical review on the current scenario and significance of crude glycerol resulting from biodiesel industry towards more sustainable renewable energy industry. Renewable and Sustainable Energy Reviews, 16, 2671–2686.CrossRefGoogle Scholar
  2. 2.
    Beopoulos, A., Cescut, J., Haddouche, R., Uribelarrea, J. L., Molina-Jouve, C., & Nicaud, J. M. (2009). Yarrowia lipolytica as a model for bio-oil production. Progress in Lipid Research, 48, 375–387.CrossRefGoogle Scholar
  3. 3.
    Beopoulos, A., Chardot, T., & Nicaud, J. M. (2009). Yarrowia lipolytica: a model and a tool to understand the mechanisms implicated in lipid accumulation. Biochimie, 91, 692–696.CrossRefGoogle Scholar
  4. 4.
    Beopoulos, A., Mrozova, Z., Thevenieau, F., Le Dall, M. T., Hapala, I., Papanikolaou, S., Chardot, T., & Nicaud, J. M. (2008). Control of lipid accumulation in the yeast Yarrowia lipolytica. Applied and Environmental Microbiology, 74, 7779–7789.CrossRefGoogle Scholar
  5. 5.
    Bondioli, P., Della, B., & Laura. (2005). An alternative spectrophotometric method for the determination of free glycerol in biodiesel. European Journal of Lipid Science and Technology, 107, 153–157.CrossRefGoogle Scholar
  6. 6.
    Burton, R., & Fan, X. (2009). Recent development of biodiesel feedstocks and the applications of glycerol: a review. The Open Fuels & Energy Science Journal, 2, 100–109.CrossRefGoogle Scholar
  7. 7.
    Campbell, C. J. (2006). The Rimini Protocol an oil depletion protocol: heading off economic chaos and political conflict during the second half of the age of oil. Energy Policy, 34, 1319–1325.CrossRefGoogle Scholar
  8. 8.
    Chatzifragkou, A., Fakas, S., Galiotou-Panayotou, M., Komaitis, M., Aggelis, G., & Papanikolaou, S. (2010). Commercial sugars as substrates for lipid accumulation in Cunninghamella echinulata and Mortierella isabellina fungi. European Journal of Lipid Science and Technology, 112, 1048–1057.CrossRefGoogle Scholar
  9. 9.
    Ciriminna, R., Pina, C. D., Rossi, M., & Pagliaro, M. (2014). Understanding the glycerol market. European Journal of Lipid Science and Technology, 116, 1432–1439.CrossRefGoogle Scholar
  10. 10.
    Cristina, V., Paulo, A., Miguel, C., & Isabel, C. (1998). The H1-ATPase in the plasma membrane of Saccharomyces cerevisiae is activated during growth latency in octanoic acid-supplemented medium accompanying the decrease in intracellular pH and cell viability. Applied and Environmental Microbiology, 64, 779–783.Google Scholar
  11. 11.
    Cui, Y., Blackburn, J. W., & Liang, Y. (2012). Fermentation optimization for the production of lipid by Cryptococcus curvatus: use of response surface methodology. Biomass and Bioenergy, 47, 410–417.CrossRefGoogle Scholar
  12. 12.
    Evans, C. T., Scragg, A. H., & Ratledge, C. (1983). A comparative study of citrate efflux from mitochondria of oleaginous and non-oleaginous yeasts. European J Biochem, 130, 195–204.CrossRefGoogle Scholar
  13. 13.
    Folch, J., Lees, M., & Slane-Stanley, G. H. (1957). Simple method for the isolation and purification of total lipids from animal tissues. The Journal of Biological Chemistry, 226, 497–509.Google Scholar
  14. 14.
    Gao, Y., Li, D., & Liu, Y. (2011). Production of single cell protein from soy molasses using Candida tropicalis. Annals of Microbiology, 62, 1165–1172.CrossRefGoogle Scholar
  15. 15.
    Giridhar, R., & Srivastava, A. K. (2000). Model based constant feed fed-batch L-sorbose production process for improvement in L-sorbose productivity. Chemical and Biochemical Engineering Quarterly, 14, 133–140.Google Scholar
  16. 16.
    Ines, S., Silla, H., Steffen, G., Thomas, R., Katrin, O., Christoph, S., & Anke, N. (2014). Characterization of newly isolated oleaginous yeasts—Cryptococcus podzolicus, Trichosporon porosum and Pichia segobiensis. AMB Express, 4, 24.CrossRefGoogle Scholar
  17. 17.
    Jarboe, L. R., Royce, L. A., & Liu, P. (2013). Understanding biocatalyst inhibition by carboxylic acids. Frontiers in Microbiology, 4, 272.CrossRefGoogle Scholar
  18. 18.
    Landman, A. D., & Dakshinamurti, K. (1975). Acetyl-Coenzyme A Carboxylase role of the prosthetic group in enzyme polymerization. Biochemical Journal, 145, 545–548.CrossRefGoogle Scholar
  19. 19.
    Lin, J., Shen, H., Tan, H., Zhao, X., Wu, S., Hu, C., & Zhao, Z. K. (2011). Lipid production by Lipomyces starkeyi cells in glucose solution without auxiliary nutrients. Journal of Biotechnology, 152, 184–188.CrossRefGoogle Scholar
  20. 20.
    Marier, J. R., & Boulet, M. (1958). Direct determination of citric acid in milk with an improved pyridine-acetic anhydride method. Journal of Dairy Sciences, 41, 1683–1692.CrossRefGoogle Scholar
  21. 21.
    Mhairi, W., Philippe, H., & Jette, T. (2013). Comparing cellular performance of Yarrowia lipolytica during growth on glucose and glycerol in submerged cultivations. AMB Express, 3, 1–9.CrossRefGoogle Scholar
  22. 22.
    Moss, J., & Lane, M. D. (1972). Acetyl Coenzyme A Carboxylase. The Journal of Biological Chemistry, 247, 4952–4959.Google Scholar
  23. 23.
    Narlin, B., Beaty, S., & Lane, M. D. (1983). Kinetics of activation of acetyl-coA Carboxylase by citrate relationship to the rate of polymerization of the enzyme. The Journal of Biological Chemistry, 258, 13043–13050.Google Scholar
  24. 24.
    Papanikolaou, S., & Aggelis, G. (2003). Selective uptake of fatty acids by the yeast Yarrowia lipolytica. European Journal of Lipid Science and Technology, 105, 651–655.CrossRefGoogle Scholar
  25. 25.
    Papanikolaou, S., Chevalot, I., Komaitis, M., & Geor. (2001). Kinetic profile of the cellular lipid composition in an oleaginous Yarrowia lipolytica capable of producing a cocoa-butter substitute from industrial fats. Antonie Van Leeuwenhoek, 80, 215–224.CrossRefGoogle Scholar
  26. 26.
    Papanikolaou, S., Chevalot, I., Komaitis, M., Marc, I., & Aggelis, G. (2002). Single cell oil production by Yarrowia lipolytica growing on an industrial derivative of animal fat in batch cultures. Applied Microbiology and Biotechnology, 58, 308–312.CrossRefGoogle Scholar
  27. 27.
    Papanikolaou, S., Muniglia, L., Chevalot, I., Aggelis, G., & Marc, I. (2002). Yarrowia lipolytica as a potential producer of citric acid from raw glycerol. Journal of Applied Microbiology, 92, 737–744.CrossRefGoogle Scholar
  28. 28.
    Ratledge, C. (2004). Fatty acid biosynthesis in microorganisms being used for single cell oil production. Biochimie, 86, 807–815.CrossRefGoogle Scholar
  29. 29.
    Ratledge, C., & Wynn, J. P. (2002). The biochemistry and molecular biology of lipid accumulation in oleaginous microorganisms. Advances in Applied Microbiology, 51, 1–51.CrossRefGoogle Scholar
  30. 30.
    Rywińska, A., Juszczyk, P., Wojtatowicz, M., Robak, M., Lazar, Z., Tomaszewska, L., & Rymowicz, W. (2013). Glycerol as a promising substrate for Yarrowia lipolytica biotechnological applications. Biomass and Bioenergy, 48, 148–166.CrossRefGoogle Scholar
  31. 31.
    Savaliya, M. L., Dhorajiya, B. D., & Dholakiya, B. Z. (2013). Recent advancement in production of liquid biofuels from renewable resources: a review. Research on Chemical Intermediates, 41, 475–509.CrossRefGoogle Scholar
  32. 32.
    Seo, Y. H., Lee, I., Jeon, S. H., & Han, J.-I. (2014). Efficient conversion from cheese whey to lipid using Cryptococcus curvatus. Biochemical Engineering Journal, 90, 149–153.CrossRefGoogle Scholar
  33. 33.
    Tomaszewska, L., Rakicka, M., Rymowicz, W., & Rywinska, A. (2014). A comparative study on glycerol metabolism to erythritol and citric acid in Yarrowia lipolytica yeast cells. FEMS Yeast Research, 14, 966–976.CrossRefGoogle Scholar
  34. 34.
    Ullah, A., Orij, R., Brul, S., & Smits, G. J. (2012). Quantitative analysis of the modes of growth inhibition by weak organic acids in Saccharomyces cerevisiae. Applied and Environmental Microbiology, 78, 8377–8387.CrossRefGoogle Scholar
  35. 35.
    Van Leeuwen, J., Rasmussen, M. L., Sankaran, S., Koza, C. R., Erickson, D. T., Mitra, D., & Jin, B. (2012). Fungal treatment of crop processing wastewaters with value-added co-products. Sustainable Bioenergy and Bioproducts, Green Energy and Technology, 8, 13–44.CrossRefGoogle Scholar
  36. 36.
    Wynn, J. P., bin Abdul Hamidt, A., & Ratledge, C. (1999). The role of malic enzyme in the regulation of lipid accumulation in filamentous fungi. Microbiology, 145, 1911–1917.CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2017

Authors and Affiliations

  1. 1.Institut National de la Recherche Scientifique, Centre Eau Terre EnvironnementUniversité du QuébecQuebec CityCanada

Personalised recommendations