Skip to main content

Dynamics of yeast immobilized-cell fluidized-bed bioreactors systems in ethanol fermentation from lactose-hydrolyzed whey and whey permeate

Abstract

We studied the dynamics of ethanol production on lactose-hydrolyzed whey (LHW) and lactose-hydrolyzed whey permeate (LHWP) in batch fluidized-bed bioreactors using single and co-cultures of immobilized cells of industrial strains of Saccharomyces cerevisiae and non-industrial strains of Kluyveromyces marxianus. Although the co-culture of S. cerevisiae CAT-1 and K. marxianus CCT 4086 produced two- to fourfold the ethanol productivity of single cultures of S. cerevisiae, the single cultures of the K. marxianus CCT 4086 produced the best results in both media (Y EtOH/S = 0.47–0.49 g g−1 and Q P = 1.39–1.68 g L−1 h−1, in LHW and LHWP, respectively). Ethanol production on concentrated LHWP (180 g L−1) reached 79.1 g L−1, with yields of 0.46 g g−1 for K. marxianus CCT 4086 cultures. Repeated batches of fluidized-bed bioreactor on concentrated LHWP led to increased ethanol productivity, reaching 2.8 g L−1 h−1.

This is a preview of subscription content, access via your institution.

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

References

  1. Guimaraes P, Teixeira J, Domingues L (2010) Fermentation of lactose to bio-ethanol by yeasts as part of integrated solutions for the valorisation of cheese whey. Biotechnol Adv 28(3):375–384

    Article  CAS  Google Scholar 

  2. Ozmihci S, Kargi F (2009) Fermentation of cheese whey powder solution to ethanol in a packed-column bioreactor: effects of feed sugar concentration. J Chem Technol Biotechnol 84(1):106–111

    Article  CAS  Google Scholar 

  3. Siso MIG (1996) The biotechnological utilization of cheese whey: a review. Bioresour Technol 57(1):1–11

    Article  Google Scholar 

  4. Dragone G, Mussatto SI, Silva J, Teixeira JA (2011) Optimal fermentation conditions for maximizing the ethanol production by Kluyveromyces fragilis from cheese whey powder. Biomass Bioenergy 35(5):1977–1982

    Article  CAS  Google Scholar 

  5. Rubio-Texeira M (2006) Endless versatility in the biotechnological applications of Kluyveromyces LAC genes. Biotechnol Adv 24(2):212–225

    Article  CAS  Google Scholar 

  6. Guimarães PMR, Teixeira JA, Domingues L (2008) Fermentation of high concentrations of lactose to ethanol by engineered flocculent Saccharomyces cerevisiae. Biotechnol Lett 30(11):1953–1958

    Article  CAS  Google Scholar 

  7. Lewandowska M, Kujawski W (2007) Ethanol production from lactose in a fermentation/pervaporation system. J Food Eng 79(2):430–437

    Article  CAS  Google Scholar 

  8. Tahoun MK, El-Nemr TM, Shata OH (1999) Ethanol from lactose in salted cheese whey by recombinant Saccharomyces cerevisiae. Zeitschrift Fur Lebensmittel-Untersuchung Und-Forschung a- Food Res Technol 208(1):60–64

    Article  CAS  Google Scholar 

  9. Grosova Z, Rosenberg M, Rebros M (2008) Perspectives and applications of immobilised beta-galactosidase in food industry—a review. Czech J Food Sci 26(1):1–14

    CAS  Google Scholar 

  10. Rubio-Texeira M (2005) A comparative analysis of the genetic switch between not-so-distant cousins: Saccharomyces cerevisiae versus Kluyveromyces marxianus. FEMS Yeast Res 5(12):1115–1128

    Article  CAS  Google Scholar 

  11. Kourkoutas Y, Bekatorou A, Banat I, Marchant R, Koutinas A (2004) Immobilization technologies and support materials suitable in alcohol beverages production: a review. Food Microbiol 21(4):377–397

    Article  CAS  Google Scholar 

  12. Verbelen P, De Schutter D, Delvaux F, Verstrepen K, Delvaux F (2006) Immobilized yeast cell systems for continuous fermentation applications. Biotechnol Lett 28(19):1515–1525

    Article  CAS  Google Scholar 

  13. Nigam J, Gogoi B, Bezbaruah R (1998) Alcoholic fermentation by agar-immobilized yeast cells. World J Microbiol Biotechnol 14(3):457–459

    Article  CAS  Google Scholar 

  14. Christensen AD, Kadar Z, Oleskowicz-Popiel P, Thomsen MH (2011) Production of bioethanol from organic whey using Kluyveromyces marxianus. J Ind Microbiol Biotechnol 38(2):283–289

    Article  CAS  Google Scholar 

  15. Gabardo S, Rech R, Ayub MAZ (2012) Performance of different immobilized-cell systems to efficiently produce ethanol from whey: fluidized batch, packed-bed and fluidized continuous bioreactors. J Chem Technol Biotechnol 87(8):1194–1201

    Article  CAS  Google Scholar 

  16. Najafpour G, Younesi H, Ismail K (2004) Ethanol fermentation in an immobilized cell reactor using Saccharomyces cerevisiae. Bioresour Technol 92(3):251–260

    Article  CAS  Google Scholar 

  17. Yu JL, Zhang X, Tan TW (2007) An novel immobilization method of Saccharomyces cerevisiae to sorghum bagasse for ethanol production. J Biotechnol 129(3):415–420

    Article  CAS  Google Scholar 

  18. Kumar S, Singh SP, Mishra IM, Adhikari DK (2011) Continuous ethanol production by Kluyveromyces sp. IIPE453 immobilized on bagasse chips in packed bed reactor. Petroleum Technol Alternative Fuels 2(1):1–6

  19. Lins A, Leao M (2002) Removal of skim milk lactose by fermentation using free and immobilized Kluyveromyces marxianus cells. World J Microbiol Biotechnol 18(3):187–192

    Article  CAS  Google Scholar 

  20. Basso LC, de Amorim HV, de Oliveira AJ, Lopes ML (2008) Yeast selection for fuel ethanol production in Brazil. FEMS Yeast Res 8(7):1155–1163

    Article  CAS  Google Scholar 

  21. Furlan SA, Carvalho-Jonas MF, Merkle R, Bértoli GB, Jonas R (1995) Aplicação do sistema Microtiter Reader na seleção de microrganismos produtores de ß galactosidase. Brazilian Arch Biol Technol 38(4):1261–1268

    CAS  Google Scholar 

  22. Gabardo S, Rech R, Rosa CA, Ayub MAZ (2014) Dynamics of ethanol production from whey and whey permeate by immobilized strains of Kluyveromyces marxianus in batch and continuous bioreactors. Renew Energy 69:89–96

    Article  CAS  Google Scholar 

  23. Timson DJ (2007) Galactose metabolism in Saccharomyces cerevisiae. Dynamic Biochemistry, Process Biotechnology and and Molecular Biology In: Global Sci Book (1):63–73

  24. Parrondo J, Garcia LA, Diaz M (2009) Nutrient balance and metabolic analysis in a Kluyveromyces marxianus fermentation with lactose-added whey. Brazilian J Chem Eng 26(3):445–456

    Article  CAS  Google Scholar 

  25. Silva A, Guimares PMR, Teixeira JA, Domingues L (2010) Fermentation of deproteinized cheese whey powder solutions to ethanol by engineered Saccharomyces cerevisiae: effect of supplementation with corn steep liquor and repeated-batch operation with biomass recycling by flocculation. J Ind Microbiol Biotechnol 37(9):973–982

    Article  CAS  Google Scholar 

  26. Ramakrishnan S, Hartley BS (1993) Fermentation of lactose by yeast-cells secreting recombinant fungal lactase. Appl Env Microbiol 59(12):4230–4235

    CAS  Google Scholar 

  27. Domingues L, Dantas MM, Lima N, Teixeira JA (1999) Continuous ethanol fermentation of lactose by a recombinant flocculating Saccharomyces cerevisiae strain. Biotechnol Bioeng 64(6):692–697

    Article  CAS  Google Scholar 

  28. Marwaha SS, Kennedy JF (1984) Ethanol-production from whey permeate by immobilized yeast-cells. Enz Microb Technol 6(1):18–22

    Article  CAS  Google Scholar 

  29. Guo XW, Zhou J, Xiao DG (2010) Improved Ethanol production by mixed immobilized Cells of Kluyveromyces marxianus and Saccharomyces cerevisiae from cheese whey powder solution fermentation. Appl Biochem Biotechnol 160(2):532–538

    Article  CAS  Google Scholar 

  30. Fonseca GG, Heinzle E, Wittmann C, Gombert AK (2008) The yeast Kluyveromyces marxianus and its biotechnological potential. Appl Microbiol Biotechnol 79(3):339–354

    Article  CAS  Google Scholar 

  31. Gosling A, Stevens GW, Barber AR, Kentish SE, Gras SL (2010) Recent advances refining galactooligosaccharide production from lactose. Food Chem 121(2):307–318

    Article  CAS  Google Scholar 

  32. Bellaver LH, de Carvalho NMB, Abrahao-Neto J, Gombert AK (2004) Ethanol formation and enzyme activities around glucose-6-phosphate in Kluyveromyces marxianus CBS 6556 exposed to glucose or lactose excess. FEMS Yeast Res 4(7):691–698

    Article  CAS  Google Scholar 

  33. Kargi F, Ozmihci S (2006) Utilization of cheese whey powder (CWP) for ethanol fermentations: effects of operating parameters. Enz Microb Technol 38(5):711–718

    Article  CAS  Google Scholar 

  34. Silveira WB, Passos F, Mantovani HC, Passos FML (2005) Ethanol production from cheese whey permeate by Kluyveromyces marxianus UFV-3: a flux analysis of oxido-reductive metabolism as a function of lactose concentration and oxygen levels. Enz Microb Technol 36(7):930–936

    Article  CAS  Google Scholar 

  35. Patil A, Koolwal S, Butala H (2002) Fusel oil: composition, removal and potencial utilization. Int Sugar J 104:51–58

    CAS  Google Scholar 

  36. Solmaz H (2015) Combustion, performance and emission characteristics of fusel oil in a spark ignition engine. Fuel Proces Technol 133:20–28

    Article  CAS  Google Scholar 

  37. Mayer FD, Feris LA, Marcilio NR, Hoffmann R (2015) Why small-scale fuel ethanol production in Brazil does not take off? Renew Sustain Energy Rev 43:687–701

    Article  Google Scholar 

  38. Sakurai A, Nishida Y, Saito H, Sakakibara M (2000) Ethanol production by repeated batch culture using yeast cells immobilized within porous cellulose carriers. J J Biosci Bioeng 90(5):526–529

    Article  CAS  Google Scholar 

  39. Athanasiadis I, Boskou D, Kanellaki M, Koutinas AA (2001) Effect of carbohydrate substrate on fermentation by kefir yeast supported on delignified cellulosic materials. J J Agric Food Chem 49(2):658–663

    Article  CAS  Google Scholar 

  40. Abe A, Furukawa S, Watanabe S, Morinaga Y (2013) Yeasts and lactic acid bacteria mixed-specie biofilm formation is a promising cell immobilization technology for ethanol fermentation. Appl Biochem Biotechnol 171(1):72–79

    Article  CAS  Google Scholar 

  41. Fan C, Qi K, Xia XX, Zhong JJ (2013) Efficient ethanol production from corncob residues by repeated fermentation of an adapted yeast. Bioresour Technol 136:309–315

    Article  CAS  Google Scholar 

  42. Watanabe I, Miyata N, Ando A, Shiroma R, Tokuyasu K, Nakamura T (2012) Ethanol production by repeated-batch simultaneous saccharification and fermentation (SSF) of alkali-treated rice straw using immobilized Saccharomyces cerevisiae cells. Bioresour Technol 123:695–698

    Article  CAS  Google Scholar 

  43. Rattanapan A, Limtong S, Phisalaphong M (2011) Ethanol production by repeated batch and continuous fermentations of blackstrap molasses using immobilized yeast cells on thin-shell silk cocoons. Appl Energy 88(12):4400–4404

    Article  CAS  Google Scholar 

  44. Ling K (2008) Whey to ethanol: a biofuel role for dairy cooperatives? USDA Rural Development (Report 214):Washington

Download references

Acknowledgments

The authors wish to thank CNPq and CAPES (Brazil) for the financial support of this research and scholarships (first and second authors).

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Marco Antônio Záchia Ayub.

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Gabardo, S., Pereira, G.F., Klein, M.P. et al. Dynamics of yeast immobilized-cell fluidized-bed bioreactors systems in ethanol fermentation from lactose-hydrolyzed whey and whey permeate. Bioprocess Biosyst Eng 39, 141–150 (2016). https://doi.org/10.1007/s00449-015-1498-0

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00449-015-1498-0

Keywords

  • Ethanol
  • Lactose-hydrolyzed whey
  • Lactose-hydrolyzed whey permeate
  • Immobilized cells
  • Saccharomyces cerevisiae
  • Kluyveromyces marxianus