Bioprocess and Biosystems Engineering

, Volume 35, Issue 6, pp 993–1004 | Cite as

Effects of some inhibitors on the growth and lipid accumulation of oleaginous yeast Rhodosporidium toruloides and preparation of biodiesel by enzymatic transesterification of the lipid

  • Xuebing ZhaoEmail author
  • Feng Peng
  • Wei Du
  • Canming Liu
  • Dehua LiuEmail author
Original Paper


Microbial lipid produced using yeast fermentation with inexpensive carbon sources such as lignocellulosic hydrolyzate can be an alternative feedstock for biodiesel production. Several inhibitors that can be generated during acid hydrolysis of lignocellulose were added solely or together into the culture medium to study their individual inhibitory actions and their synergistic effects on the growth and lipid accumulation of oleaginous yeast Rhodosporidium toruloides. When the inhibitors were present in isolation in the medium, to obtain a high cell biomass accumulation, the concentrations of formic acid, acetic acid, furfural and vanillin should be lower than 2, 5, 0.5 and 1.5 g/L, respectively. However, the synergistic effects of these compounds could dramatically decrease the minimum critical inhibitory concentrations leading to significant growth and lipid production inhibitions. Unlike the above-cited inhibitors, sodium lignosulphonate had no negative influence on biomass accumulation when its concentration was in the range of 0.5–2.0 g/L; in effect, it was found to facilitate cell growth and sugar-to-lipid conversion. The fatty acid compositional profile of the yeast lipid was in the compositional range of various plant oils and animal tallow. Finally, the crude yeast lipid from bagasse hydrolyzate could be well converted into fatty acid methyl ester (FAME, biodiesel) by enzymatic transesterification in a tert-butanol system with biodiesel yield of 67.2% and lipid-to-biodiesel conversion of 88.4%.


Yeast lipid Lignocellulosic biomass Detoxification of hydrolyzate Inhibition Biodiesel 



This work was supported by Tsinghua Research Funding (No. 2009THZ0223). The authors sincerely appreciate Mr. Ilan E. Cuperstein, Dr. Quin Garcia and Dr. Erik Chavez for their help in polishing English language.


  1. 1.
    Boyer LJ, Vega JL, Klasson KT, Clausen EC, Gaddy JL (1992) The effects of furfural on ethanol production by Saccharomyces cerevisiae in batch culture. Biomass Bioenergy 3:41–48CrossRefGoogle Scholar
  2. 2.
    Delgenes J, Moletta R, Navarro JM (1996) Effects of lignocellulose degradation products on ethanol fermentations of glucose and xylose by Saccharomyces cerevisiae, Zymomonas mobilis, Pichia stipitis and Candida shehatae. Enzyme Microb Technol 19:220–225CrossRefGoogle Scholar
  3. 3.
    Fjerbaek L, Christensen KV, Norddahl B (2009) A review of the current state of biodiesel production using enzymatic transesterification. Biotechnol Bioeng 102:1298–1315CrossRefGoogle Scholar
  4. 4.
    Guo GL, Chen WH, Chen WH, Men LC, Hwang WS (2008) Characterization of dilute acid pretreatment of silvergrass for ethanol production. Bioresour Technol 99:6046–6053CrossRefGoogle Scholar
  5. 5.
    Hu C, Zhao X, Zhao J, Wu S, Zhao ZK (2009) Effects of biomass hydrolysis by-products on oleaginous yeast Rhodosporidium toruloides. Bioresour Technol 100:4843–4847CrossRefGoogle Scholar
  6. 6.
    Hu C, Wu S, Wang Q, Zhao Z (2011) Simultaneous utilization of glucose and xylose for lipid production by Trichosporon cutaneum. Biotechnol Biofuels 4:25CrossRefGoogle Scholar
  7. 7.
    Huang C, Zong MH, Wu H, Liu QP (2009) Microbial oil production from rice straw hydrolysate by Trichosporon fermentans. Bioresour Technol 19:4535–4538CrossRefGoogle Scholar
  8. 8.
    Karmakar A, Karmakar S, Mukherjee S (2010) Properties of various plants and animals feedstocks for biodiesel production. Bioresour Technol 101:7201–7210CrossRefGoogle Scholar
  9. 9.
    Lee WG, Lee JS, Shin CS, Park SC, Chang HN, Chang YK (1999) Ethanol production using concentrated oak wood hydrolysates and methods to detoxify. Appl Biochem Biotechnol 78:547–559CrossRefGoogle Scholar
  10. 10.
    Li ZF, Zhang L, Shen XJ, Lai BS, Sun SQ (2001) A comparative study on four method of fungi lipid extraction. Microbiol 28:72–75Google Scholar
  11. 11.
    Li YH, Liu B, Sun Y, Zhao ZB, Bai FW (2005) Screening of oleaginous yeasts for broad-spectrum carbohydrates assimilation capacity. Chin Biotechnol 25:39–43Google Scholar
  12. 12.
    Li YH, Liu B, Zhao ZB (2006) Optimization of culture conditions for lipid production by Rhodosporidium toruloides. Chin J Biotechnol 22:650–656CrossRefGoogle Scholar
  13. 13.
    Li L, Du W, Liu D, Li Z (2006) Lipase-catalyzed transesterification of rapeseed oils for biodiesel production with a novel organic solvent as the reaction medium. J Mol Catal B Enzym 43:58–62CrossRefGoogle Scholar
  14. 14.
    Li YH, Zhao ZB, Bai FW (2007) High-density cultivation of oleaginous yeast Rhodosporidium toruloides Y4 in fed-batch culture. Enzyme Microb Technol 41:312–317CrossRefGoogle Scholar
  15. 15.
    Li Q, Du W, Liu D (2008) Perspectives of microbial oils for biodiesel production. Appl Microbiol Biotechnol 80:749–756CrossRefGoogle Scholar
  16. 16.
    Ma F, Hanna MA (1999) Biodiesel production: a review. Bioresour Technol 70:1–15CrossRefGoogle Scholar
  17. 17.
    Modig T, Lidén G, Taherzadeh MJ (2002) Inhibition effects of furfural on alcohol dehydrogenase, aldehyde dehydrogenase and pyruvate dehydrogenase. Biochem J 363:769–776CrossRefGoogle Scholar
  18. 18.
    Mussatto SI, Roberto JC (2004) Alternatives for detoxification of diluted-acid lignocellulosic hydrolyzates for use in fermentative processes: a review. Bioresour Technol 93:1–10CrossRefGoogle Scholar
  19. 19.
    Nigam JN (2001) Ethanol production from wheat straw hemicellulose hydrolysate by Pichia stipitis. J Biotechnol 87:17–27CrossRefGoogle Scholar
  20. 20.
    Palmqvist E, Hahn-Hagerdal B (2000) Fermentation of lignocellulosic hydrolysates. II: inhibitors and mechanisms of inhibition. Bioresour Technol 74:25–33CrossRefGoogle Scholar
  21. 21.
    Palmqvist E, Grage H, Meinander NQ, Hahn-Hägerdal B (1999) Main and interaction effects of acetic acid, furfural and p-hydroxybenzoic acid on growth and ethanol productivity of yeasts. Biotechnol Bioeng 63:46–55CrossRefGoogle Scholar
  22. 22.
    Papanikolaou S, Dimou A, Fakas S, Diamantopoulou P, Philippoussis A, Galiotou-Panayotou M, Aggelis G (2011) Biotechnological conversion of waste cooking olive oil into lipid-rich biomass using Aspergillus and Penicillium strains. J Appl Microbiol 110:1138–1150CrossRefGoogle Scholar
  23. 23.
    Parajó JC, Domnguez H, Domnguez JM (1998) Biotechnological production of xylitol. Part 3: operation in culture media made from lignocellulose hydrolysates. Bioresour Technol 66:25–40CrossRefGoogle Scholar
  24. 24.
    Pienkos PT, Zhang M (2009) Role of pretreatment and conditioning processes on toxicity of lignocellulosic biomass hydrolysates. Cellulose 16:743–762CrossRefGoogle Scholar
  25. 25.
    Saha BC, Bothast RJ (1999) Pretreatment and enzymatic saccharification of corn fiber. Appl Biochem Biotechnol 76:65–77CrossRefGoogle Scholar
  26. 26.
    Sárvári Horváth I, Taherzadeh MJ, Niklasson C, Lidén G (2001) Effects of furfural on anaerobic continuous cultivation of Saccharomyces cerevisiae. Biotechnol Bioeng 75:540–549CrossRefGoogle Scholar
  27. 27.
    Sivers MV, Zacchi G (1996) Ethanol from lignocellulosics: a review of the economy. Bioresour Technol 56:131–140CrossRefGoogle Scholar
  28. 28.
    Taherzadeh MJ, Niklasson C, Liden G (1997) Acetic acid—friend or foe in anaerobic batch conversion of glucose to ethanol by Saccharomyces cerevisiae. Chem Eng Sci 52:2653–2659CrossRefGoogle Scholar
  29. 29.
    Taoka Y, Nagano N, Okita Y, Izumida H, Sugimoto S, Hayashi M (2011) Effect of Tween 80 on the growth, lipid accumulation and fatty acid composition of Thraustochytrium aureum ATCC 34304. J Biosci Bioeng 111:420–424CrossRefGoogle Scholar
  30. 30.
    Van Zyl C, Prior B, du Preez JC (1991) Acetic acid inhibition of d-xylose fermentation by Pichia stipitis. Enzyme Microb Technol 13:82–86CrossRefGoogle Scholar
  31. 31.
    Wu S, Hu C, Jin G, Zhao X, Zhao ZK (2010) Phosphate-limitation mediated lipid production by Rhodosporidium toruloides. Bioresour Technol 101:6124–6129CrossRefGoogle Scholar
  32. 32.
    Wu S, Zhao X, Shen H, Wang Q, Zhao ZK (2011) Microbial lipid production by Rhodosporidium toruloides under sulfate-limited conditions. Bioresour Technol 102:1803–1807CrossRefGoogle Scholar
  33. 33.
    Xing D, Pan A, Xue D, Fang M, Gu R (2010) Biomass carbohydrates assimilation and lipid accumulation by Mortierella isabellina. Chin J Biotechnol 26:189–193Google Scholar
  34. 34.
    Yu X, Zheng Y, Dorgan KM, Chen S (2011) Oil production by oleaginous yeasts using the hydrolysate from pretreatment of wheat straw with dilute sulfuric acid. Bioresour Technol 102:6134–6140CrossRefGoogle Scholar
  35. 35.
    Zaldivar J, Martinez A, Ingram LO (2000) Effect of alcohol compounds found in hemicellulose hydrolysate on growth and fermentation of ethanologenic Escherichia coli. Biotechnol Bioeng 68:524–530CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2012

Authors and Affiliations

  1. 1.Institute of Applied Chemistry, Department of Chemical EngineeringTsinghua UniversityBeijingChina
  2. 2.College of ScienceHunan Agricultural UniversityChangshaChina

Personalised recommendations