Annals of Microbiology

, Volume 62, Issue 3, pp 987–993 | Cite as

Co-culture of an oleaginous yeast Rhodotorula glutinis and a microalga Chlorella vulgaris for biomass and lipid production using pure and crude glycerol as a sole carbon source

  • Benjamas Cheirsilp
  • Suleeporn Kitcha
  • Salwa Torpee
Original Article


This study has shown that a co-culture of an oleaginous yeast Rhodotorula glutinis TISTR 5159 and a microalga Chlorella vulgaris var. vulgaris TISTR 8261 enhanced biomass and lipid production from glycerol. It is possible that the microalga may function as an oxygen producer in the co-culture and enhance the growth of yeast. The use of 3% pure glycerol as a carbon source and urea as a nitrogen source with a molar carbon-to-nitrogen (C/N) ratio of 32 gave the highest biomass and lipid production. These produced a 5.7-fold and 3.8-fold of biomass and lipid, respectively, compared to the initial unoptimized condition. The co-culture system was also applied to convert crude glycerol, a by-product from a biodiesel plant, to biomass and lipid. The lipid produced from the crude glycerol by the co-culture was mainly composed of palmitic acid (C16:0) 40.52% and oleic acid (C18:1) 21.30%, which was a plant oil-like fatty acid composition. This suggests that it has a high potential to be used as a biodiesel feedstock.


Biodiesel Glycerol Lipid Microalgae Oleaginous yeast 


  1. Angerbauer C, Siebenhofer M, Mittelbach M, Guebitz GM (2008) Conversion of sewage sludge into lipids by Lipomyces starkeyi for biodiesel production. Bioresour Technol 99:3051–3056PubMedCrossRefGoogle Scholar
  2. Cai SQ, Hu C, Du S (2007) Comparisons of growth and biochemical composition between mixed culture of alga and yeast and monocultures. J Biosci Bioeng 104(5):391–397PubMedCrossRefGoogle Scholar
  3. Chatzifragkou A, Makri A, Belka A, Bellou S, Mavrou M, Mastoridou M, Mystrioti P, Onjaro G, Aggelis G, Papanikolaou S (2011) Biotechnological conversions of biodiesel derived waste glycerol by yeast and fungal species. Energy 36:1097–1108CrossRefGoogle Scholar
  4. Cheirsilp B, Suwannarat W, Niyomdecha R (2011) Mixed culture of oleaginous yeast Rhodotorula glutinis and microalga Chlorella vulgaris for lipid production from industrial wastes and its use as biodiesel feedstock. New Biotechnol 28(4):362–368CrossRefGoogle Scholar
  5. Converti A, Casazza AA, Ortiz EY, Perego P, Borghi MD (2009) Effect of temperature and nitrogen concentration on the growth and lipids content of Nannochloropsis oculata and Chlorella vulgaris for biodiesel production. Chem Eng Process: Proc Intensification 48(6):1146–1151CrossRefGoogle Scholar
  6. Dasari MA, Kiatsimkul PP, Sutterlin WR, Suppes GJ (2005) Low-pressure hydrogenolysis of glycerol to propylene glycol. Appl Catal A: Gen 281:225–231CrossRefGoogle Scholar
  7. Fakas S, Galiotou-Panayotou M, Papanikolaou S, Komaitis M, Aggelis G (2007) Compositional shifts in lipid fractions during lipid turnover in Cunninghamella echinulata. Enzyme Microb Technol 40:1321–1327CrossRefGoogle Scholar
  8. Fakas S, Makri A, Bellou S, Aggelis G (2009) Pathways to aerobic glycerol catabolism and their regulation. In: Aggelis G (ed) Microbial conversions of raw glycerol. Nova Science, New York, pp 9–18Google Scholar
  9. Folch J, Lees M, Stanley GHS (1957) A simple method for the isolation and purification of total lipides from animal tissues. J Biol Chem 226:497–509PubMedGoogle Scholar
  10. Illman AM, Scragg AH, Shales SW (2000) Increase in Chlorella strains calorific values when grown in low nitrogen medium. Enzyme Microb Technol 27:631–635PubMedCrossRefGoogle Scholar
  11. Jham GN, Teles FFF, Campos LG (1982) Use of aqueous HCl/MeOH as esterification reagent for analysis of fatty acids derived from soybean lipids. J Am Oil Chem Soc 59(3):132–133CrossRefGoogle Scholar
  12. Kosugi Y, Igusa H, Tomizuka N (1994) Continual conversion of free fatty acid in rice bran oil to triacylglycerol by immobilized lipase. J Jpn Oil Chem Soc 71:445–448Google Scholar
  13. Largeau C, Casadevall E, Berkaloff C, Dhamelincourt P (1980) Sites of accumulation and composition of hydrocarbons in Botryococcus braunii. Phytochemistry 19:1043–1051CrossRefGoogle Scholar
  14. Li Q, Wang MY (1997) Use food industry waste to produce microbial oil. Sci Technol Food Ind 6:65–69Google Scholar
  15. Li Y, Zhao Z, Bai F (2007) High–density cultivation of oleaginous yeast Rhodosporidium toruloides Y4 in fed–batch culture. Enzyme Microb Technol 41:312–317CrossRefGoogle Scholar
  16. Li M, Liu GL, Chi Z, Chi ZM (2010) Single cell oil production from hydrolysate of cassava starch by marine-derived yeast Rhodotorula mucilaginosa TJY15a. Biomass Bioenergy 34:101–107CrossRefGoogle Scholar
  17. Liang Y, Sarkany N, Cui Y (2009) Biomass and lipid productivities of Chlorella vulgaris under autotrophic, heterotrophic and mixotrophic growth conditions. Biotechnol Lett 31:1043–1049PubMedCrossRefGoogle Scholar
  18. Liang Y, Cui Y, Trushenski J, Blackburn JW (2010) Converting crude glycerol derived from yellow grease to lipids through yeast fermentation. Bioresour Technol 101:7581–7586PubMedCrossRefGoogle Scholar
  19. Makri A, Fakas S, Aggelis G (2010) Metabolic activities of biotechnological interest in Yarrowia lipolytica grown on glycerol in repeated batch cultures. Bioresour Technol 101:2351–2358PubMedCrossRefGoogle Scholar
  20. Meesters PAEP, Huijberts GNM, Eggink G (1996) High-cell-density cultivation of the lipid accumulating yeast Cryptococcus curvatus using glycerol as a carbon source. Appl Microbiol Biotechnol 45:575–579CrossRefGoogle Scholar
  21. Papanikolaou S, Aggelis G (2002) Lipid production by Yarrowia lipolytica growing on industrial glycerol in a single-stage continuous culture. Bioresour Technol 82:43–49PubMedCrossRefGoogle Scholar
  22. Papanikolaou S, Aggelis G (2009) Biotechnological valorisation of biodiesel derived glycerol waste through production of single cell oil and citric acid by Yarrowia lipolytica. Lipid Technol 21:83–87CrossRefGoogle Scholar
  23. Papanikolaou S, Sarantou S, Komaitis M, Aggelis G (2004) Repression of reserve lipid turnover in Cunninghamella echinulata and Mortierella isabellina cultivated in multiple-limited media. J Appl Microbiol 97:867–875PubMedCrossRefGoogle Scholar
  24. Papanikolaou S, Fakas S, Fick M, Chevalot I, Galiotou-Panayotou M, Komaitis M, Marc I, Aggelis G (2008) Biotechnological valorisation of raw glycerol discharged after bio-diesel (fatty acid methyl esters) manufacturing process: Production of 1,3-propanediol, citric acid and single cell oil. Biomass Bioenergy 32:60–71CrossRefGoogle Scholar
  25. Petkov G, Garcia G (2007) Which are fatty acids of the green alga Chlorella? Biochem Syst Ecol 35(5):281–285CrossRefGoogle Scholar
  26. Saenge C, Cheirsilp B, Suksaroge TT, Bourtoom T (2010) Potential use of oleaginous red yeast Rhodotorula glutinis for the bioconversion of crude glycerol from biodiesel plant to lipids and carotenoids. Process Biochem 46(1):210–218CrossRefGoogle Scholar
  27. Saenge C, Cheirsilp B, Suksaroge T, Bourtoom T (2011) Efficient concomitant production of lipids and carotenoids by oleaginous red yeast Rhodotorula glutinis cultured in palm oil mill effluent and application of lipids for biodiesel production. Biotechnol Bioprocess Eng 16(1):23–33CrossRefGoogle Scholar
  28. Shia XM, Zhang XW, Chen F (2000) Heterotrophic production of biomass and lutein by Chlorella protothecoides on various nitrogen sources. Enzyme Microb Technol 27:312–318CrossRefGoogle Scholar
  29. Xue F, Miao J, Zhang X, Tan T (2010) A new strategy for lipids production by mix cultivation of Spirulina platensis and Rhodotorula glutinis. Appl Biochem Biotechnol 160(2):498–503PubMedCrossRefGoogle Scholar
  30. Zhao CH, Zhang T, Li M, Chi ZM (2010) Single cell oil production from hydrolysates of inulin and extract of tubers of Jerusalem artichoke by Rhodotorula mucilaginosa TJY15a. Process Biochem 45:1121–1126CrossRefGoogle Scholar

Copyright information

© Springer-Verlag and the University of Milan 2011

Authors and Affiliations

  • Benjamas Cheirsilp
    • 1
  • Suleeporn Kitcha
    • 1
  • Salwa Torpee
    • 1
  1. 1.Department of Industrial Biotechnology, Faculty of Agro-IndustryPrince of Songkla UniversityHat-YaiThailand

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