Export file:


  • RIS(for EndNote,Reference Manager,ProCite)
  • BibTex
  • Text


  • Citation Only
  • Citation and Abstract

Investigating the potential of thermophilic species for ethanol production from industrial spent sulfite liquor

1 Kompetenzzentrum Holz GmbH, Altenbergerstraße 69, 4040 Linz, Austria;
2 Vienna University of Technology, Institute of Chemical Engineering Research Area Biochemical Engineering, Gumpendorferstraße 1A, 1060 Vienna, Austria;
3 Vienna University of Technology, Institute of Chemical Engineering Research Area Chemical Engineering, Getreidemarkt 9, 1060 Vienna, Austria

Special Issues: Advances in Production of Biofuels

Thermophilic microorganisms hold a great potential for bioethanol production on waste biomass, due to their ability to utilize pentoses and hexoses alike. However, to date hardly any data on thermophiles growing directly on industrial substrates like spent sulfite liquor (SSL) are available. This contribution investigates the ability of Thermoanaerobacter species to utilize the main sugars in the used SSL (mannose, glucose and xylose) and the effect of process parameters (pH, temperature and sugar concentration) on their growth. Based on these results the strain T. mathranii was chosen for further studies. The ability of T. mathranii to grow directly on SSL was investigated and the effect of several inhibiting substances on growth was elucidated. Furthermore it was tested whether pretreatment with activated charcoal can increase the fermentability of SSL. The fermentations were evaluated based on yields and specific rates. It could be shown that T. mathranii was able to ferment all sugars in the investigated softwood SSL and fermented diluted, untreated SSL (up to 2.7% (w/w) dry matter). Pretreatment with activated charcoal could slightly reduce the amount of phenols in the substrate and thus facilitate growth and ethanol production on higher SSL concentrations (up to 4.7% (w/v) dry matter). Ethanol yields of 0.29-0.44 Cmmol of ethanol per Cmmol sugar were obtained on untreated and pretreated spent sulfite liquor, respectively. These results on an industrial substrate strengthen the claim that thermophilic microorganisms might be the optimal candidates for forest biorefinery.
  Article Metrics


1. Maniatis K, Chiaramonti D, Thornley P (2012) Framework and perspectives of industrial lignocellulosic ethanol deployment: Introduction to the 1st International Conference on Lignocellulosic Ethanol. Biomass Bioenerg 46: 1-4.    

2. Fernandes DLA, Pereira SR, Serafim LS, et al. (2012) Second Generation Bioethanol from Lignocellulosics: Processing of Hardwood Sulphite Spent Liquor. In: Pinheiro Lima MA, editor. Bioethanol: InTech. pp. 123-152.

3. Nigam JN (2001) Ethanol production from hardwood spent sulfite liquor using an adapted strain of Pichia stipitis. J Ind Microbiol Biotechnol 26: 145-150.    

4. Frederick WJ, Lien SJ, Courchene CE, et al. (2008) Co-production of ethanol and cellulose fiber from Southern Pine: A technical and economic assessment. Biomass Bioenerg 32: 1293-1302    

5. Rødsrud G (2011) Biorefining-expanding the sulfite pulping biorefinery concept; Conference presentation at World Biofuels Markets; Rotterdam.

6. Lawford HG, Rousseau JD (1993) Production of ethanol from pulp mill hardwood and softwood spent sulfite liquors by genetically engineered E. coli. Appl Biochem Biotechnol 39-40: 667-685.    

7. Helle SS, Lin T, Duff SJB (2008) Optimization of spent sulfite liquor fermentation. Enzyme Microb Technol 42: 259-264.    

8. Batt CA, Caryallo S, Easson DD, et al. (1986) Direct evidence for a xylose metabolic pathway in Saccharomyces cerevisiae. Biotechnol Bioeng 28: 549-553.    

9. Pereira S, Ivanusa S, Evtuguin D, et al. (2012) Biological treatment of eucalypt spent sulphite liquors: a way to boost the production of second generation bioethanol. Biores Technol 103: 131-135.    

10. Björling T, Lindman B (1989) Evaluation of xylose-fermenting yeasts for ethanol production from spent sulfite liquor. Enzyme Microb Technol 11: 240-246.    

11. Kurtzman C, Suzuki M (2010) Phylogenetic analysis of ascomycete yeasts that form coenzyme Q-9 and the proposal of the new genera Babjeviella, Meyerozyma, Millerozyma, Priceomyces, and Scheffersomyces. Mycoscience 51: 2-14.    

12. Yu S, Wayman M, Parekh SK (1987) Fermentation to ethanol of pentose-containing spent sulphite liquor. Biotechnol Bioeng 29: 1144-1150.    

13. Lai LX (2010) Bioproducts from sulfite pulping: Bioconversion of sugar streams from pulp, sludge, and spent sulfite liquor: University of Washington.

14. Taherzadeh MJ, Fox M, Hjorth H, et al. (2003) Production of mycelium biomass and ethanol from paper pulp sulfite liquor by Rhizopus oryzae. Bioresour Technol 88: 167-177.    

15. Lindén T, Hahn-Hägerdal B (1989) Fermentation of lignocellulose hydrolysates with yeasts and xylose isomerase. Enzyme Microb Technol 11: 583-589.    

16. Helle SS, Murray A, Lam J, et al. (2004) Xylose fermentation by genetically modified Saccharomyces cerevisiae 259ST in spent sulfite liquor. Bioresour Technol 92: 163-171.    

17. Guo Z, Olsson L (2014) Characterization and fermentation of side streams from sulfite pulping. Process Biochem 49: 1231-1237.    

18. Pinel D, D'Aoust F, del Cardayre SB, et al. (2011) Saccharomyces cerevisiae Genome Shuffling through Recursive Population Mating Leads to Improved Tolerance to Spent Sulfite Liquor. App Env Microbiol 77: 4736-4743.    

19. Pereira S, Sanchez i Nogue V, Frazao C, et al. (2015) Adaptation of Scheffersomyces stipitis to hardwood spent sulfite liquor by evolutionary engineering. Biotechnol Biofuel 8: 50.    

20. Steensels J, Snoek T, Meersman E, et al. (2014) Improving industrial yeast strains: exploiting natural and artificial diversity. FEMS Microbiol Rev 38: 947-995.    

21. Boyer LJ, Vega JL, Klasson KT, et al. (1992) The effects of furfural on ethanol production by saccharomyces cereyisiae in batch culture. Biomass Bioenerg 3: 41-48.    

22. Palmqvist E, Almeida JS, Hahn-Hagerdal B (1999) Influence of furfural on anaerobic glycolytic kinetics of Saccharomyces cerevisiae in batch culture. Biotechnol Bioeng 62: 447-454.

23. Olsson L, Hahn-Hägerdal B (1993) Fermentative performance of bacteria and yeasts in lignocellulose hydrolysates. Process Biochem 28: 249-257.    

24. Parajó JC, Domínguez H, Domínguez J (1998) Biotechnological production of xylitol. Part 3: Operation in culture media made from lignocellulose hydrolysates. Bioresour Technol 66: 25-40.

25. Strickland RC, Beck MJ (1984) Effective pretreatments and neutralization methods for ethanol production from acid - catalyzed hardwood hydrolyzates using Pachysolen tannophilus. Muscle Shoals: T.V.A.

26. Xavier A, Correia M, Pereira S, et al. (2010) Second-generation bioethanol from eucalypt sulphite spent liquor. Bioresour Technol 101: 2755-2761.    

27. Bajwa PK, Shireen T, D'Aoust F, et al. (2009) Mutants of the pentose-fermenting yeast Pichia stipitis with improved tolerance to inhibitors in hardwood spent sulfite liquor. Biotechnol Bioeng 104: 892-900.    

28. Lynd LR, Cushman JH, Nichols RJ, et al. (1991) Fuel Ethanol from Cellulosic Biomass. Science 251: 1318-1323.    

29. Payton MA (1984) Production of ethanol by thermophilic bacteria. Trends Biotechnol 2: 153-158.    

30. Klinke HB, Thomsen AB, Ahring BK (2004) Inhibition of ethanol-producing yeast and bacteria by degradation products produced during pre-treatment of biomass. Appl Microbiol Biotechnol 66: 10-26.    

31. Klinke H, Thomsen A, Ahring B (2001) Potential inhibitors from wet oxidation of wheat straw and their effect on growth and ethanol production by Thermoanaerobacter mathranii. Appl Microbiol Biotechnol 57: 631-638.    

32. Lynd LR, Baskaran S, Casten S (2001) Salt Accumulation Resulting from Base Added for pH Control, and Not Ethanol, Limits Growth of Thermoanaerobacterium thermosaccharolyticum HG-8 at Elevated Feed Xylose Concentrations in Continuous Culture. Biotechnol Prog 17: 118-125.    

33. Ng TK, Ben-Bassat A, Zeikus JG (1981) Ethanol Production by Thermophilic Bacteria: Fermentation of Cellulosic Substrates by Cocultures of Clostridium thermocellum and Clostridium thermohydrosulfuricum. Appl Environ Microb 41: 1337-1343.

34. Svetlitchnyi VA, Kensch O, Falkenhan DA, et al. (2013) Single-step ethanol production from lignocellulose using novel extremely thermophilic bacteria. Biotechnol Biofuels 6: 31.    

35. Georgieva T, Ahring B (2007) Evaluation of continuous ethanol fermentation of dilute-acid corn stover hydrolysate using thermophilic anaerobic bacterium Thermoanaerobacter BG1L1. Appl Microbiol Biotechnol 77: 61-68.    

36. Georgieva T, Mikkelsen M, Ahring B (2008) Ethanol Production from Wet-Exploded Wheat Straw Hydrolysate by Thermophilic Anaerobic Bacterium Thermoanaerobacter BG1L1 in a Continuous Immobilized Reactor. In: Adney W, McMillan J, Mielenz J et al., editors. Biotechnology for Fuels and Chemicals: Humana Press. 99-110.

37. Rittmann S, Herwig C (2012) A comprehensive and quantitative review of dark fermentative biohydrogen production. Microb Cell Fact 11: 115.    

38. Wold S, Sjöström M, Eriksson L (2001) PLS-regression: a basic tool of chemometrics. Chemometr Intell Lab 58: 109-130.    

39. Grant TM, King CJ (1990) Mechanism of irreversible adsorption of phenolic compounds by activated carbons. Ind Eng Chem Res 29: 264-271.    

40. De Sousa F, Reimann A, Björklund M, et al. (2001) Estimating the amount of phenolic hydroxyl groups in lignins. Proceedings of the 11th International Symposium on Wood Pulp Chemistry 3: 649-653.

41. Weissgram M, Gstöttner J, Lorantfy B, et al. (2015) Generation of PHB from spent sulfite liquor using halophilic microorganisms. Microorganisms 3: 268.    

42. Chang T, Yao S (2011) Thermophilic, lignocellulolytic bacteria for ethanol production: current state and perspectives. Appl Microbiol Biotechnol 92: 13-27.    

43. Gütsch Jenny S, Sixta H (2011) Purification of Eucalyptus globulus water prehydrolyzates using the HiTAC process (high-temperature adsorption on activated charcoal). Holzforschung 65: 511-518.

44. Gütsch JS, Sixta H (2012) Regeneration of Spent Activated Charcoals Used for Lignin Removal from Prehydrolysis-Kraft Prehydrolyzates. Ind Eng Chem Res 51: 8624-8630.    

45. Mohan SV, Karthikeyan J (1997) Removal of lignin and tannin colour from aqueous solution by adsorption onto activated charcoal. Environ Pollut 97: 183-187.    

46. Mussatto SI, Roberto IC (2004) Alternatives for detoxification of diluted-acid lignocellulosic hydrolyzates for use in fermentative processes: a review. Biores Technol 93: 1-10.    

47. Parajó JC, Dominguez H, Domínguez JM (1997) Improved xylitol production with Debaryomyces hansenii Y-7426 from raw or detoxified wood hydrolysates. Enzyme Microb Technol 21: 18-24.    

48. Maddox IS, Murray A (1983) Production of n-butanol by fermentation of wood hydrolysate. Biotechnol Lett 5: 175-178.    

49. Montané D, Nabarlatz D, Martorell A, et al. (2006) Removal of lignin and associated impurities from xylo-oligosaccharides by activated carbon adsorption. Ind Eng Chem Res 45: 2294-2302.    

50. Barnard D, Casanueva A, Tuffin M, et al. (2010) Extremophiles in biofuel synthesis. Environ Technol 31: 871-888.    

51. Brynjarsdottir H, Wawiernia B, Orlygsson J (2012) Ethanol Production from Sugars and Complex Biomass by Thermoanaerobacter AK5: The Effect of Electron-Scavenging Systems on End-Product Formation. Energ Fuel 26: 4568-4574.    

52. Liu HS, Hsu HW, Sayler GS (1988) Bioconversion of D-Xylose and Pretreated Oak Sawdust to Ethanol Using Clostridium Thermosacchrolyticum by Batch and Continuous Up-Flow Reactors. Biotechnol Prog 4: 40-46.    

53. Helle S, Cameron D, Lam J, et al. (2003) Effect of inhibitory compounds found in biomass hydrolysates on growth and xylose fermentation by a genetically engineered strain of S. cerevisiae. Enzyme Microb Technol 33: 786-792.    

54. Carreira LH, Wiegel J, Ljungdahl LG (1983) Production of ethanol from biopolymers by anaerobic, thermophilic, and extreme thermophilic bacteria. I. Regulation of carbohydrate utilization in mutants of Thermoanaerobacter ethanolicus. Medium: X; Size: Pages: 183-191.

55. Cook GM, Janssen PH, Morgan HW (1993) Simultaneous uptake and utilisation of glucose and xylose by Clostridium thermohydrosulfuricum. FEMS Microbiology Letters 109: 55-61.    

56. Shaw AJ, Podkaminer KK, Desai SG, et al. (2008) Metabolic engineering of a thermophilic bacterium to produce ethanol at high yield. Proc Natl Acad Sci 105: 13769-13774.    

Copyright Info: © 2015, Christoph Herwig, et al., licensee AIMS Press. This is an open access article distributed under the terms of the Creative Commons Attribution Licese (http://creativecommons.org/licenses/by/4.0)

Download full text in PDF

Export Citation

Article outline

Show full outline
Copyright © AIMS Press All Rights Reserved