Archives of Microbiology

, Volume 150, Issue 1, pp 48–55 | Cite as

Ethanol production from xylose by Thermoanaerobacter ethanolicus in batch and continuous culture

  • Lynda S. Lacis
  • Hugh G. Lawford
Original Papers


The fermentation of xylose by Thermoanaerobacter ethanolicus ATCC 31938 was studied in pH-controlled batch and continuous cultures. In batch culture, a dependency of growth rate, product yield, and product distribution upon xylose concentration was observed. With 27 mM xylose media, an ethanol yield of 1.3 mol ethanol/mol xylose (78% of maximum theoretical yield) was typically obtained. With the same media, xylose-limited growth in continuous culture could be achieved with a volumetric productivity of 0.50 g ethanol/liter h and a yield of 0.42 g ethanol/g xylose (1.37 mol ethanol/mol xylose). With extended operation of the chemostat, variation in xylose uptake and a decline in ethanol yield was seen. Instability with respect to fermentation performance was attributed to a selection for mutant populations with different metabolic characteristics. Ethanol production in these T. ethanolicus systems was compared with xylose-to-ethanol conversions of other organisms. Relative to the other systems, T. ethanolicus offers the advantages of a high ethanol yield at low xylose concentrations in batch culture and of a rapid growth rate. Its disadvantages include a lower ethanol yield at higher xylose concentrations in batch culture and an instability of fermentation characteristics in continuous culture.

Key words

Thermoanaerobacter ethanolicus Ethanol Continuous fermentation Mutant selection Xylose fermentation 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Assarson P, Holysh M, Tallevi A, Wayman M, Parekh S (1986) Acid hydrolysis of lignocellulosics for alcohol production. Preprint, Can Chem Eng Conference, Sarnia, CanadaGoogle Scholar
  2. Ben-Bassat A, Lamed R, Zeikus JG (1981) Ethanol production by thermophilic bacteria: metabolic control of end product formation in Thermoanaerobium brockii. J Bacteriol 146:192–199Google Scholar
  3. Cameron DC, Cooney CL (1986) A novel fermentation: the production of R(-)-1,2-propanediol and acetol by Clostridium thermosaccharolyticum. Bio/Technology 4:651–654Google Scholar
  4. Carreira LH, Ljungdahl LG (1983) Production of ethanol from biomass using anaerobic thermophilic bacteria. In: Wise DL (ed) Liquid fuel developments. CRC Press Inc., Boca Raton, FL, pp 1–29Google Scholar
  5. 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. Biotechnol Bioeng Symp 13:183–191Google Scholar
  6. Chan E-C, Ueng PP, Chen L (1986) D-Xylose fermentation to ethanol by Schizosaccharomyces pombe cloned with xylose isomerase gene. Biotechnol Lett 8:231–234Google Scholar
  7. Clark T, Wedlock N, James AP, Deverall K, Thornton RJ (1986) Strain improvement of the xylose-fermenting yeast Pachysolen tannophilus by hybridisation of two mutant strains. Biotechnol Lett 8:801–806Google Scholar
  8. Cocks GT, Aguilar J, Lin ECC (1974) Evolution of L-1,2-propanediol catabolism in Escherichia coli by recruitment of enzymes for L-fucose and L-lactate metabolism. J Bacteriol 118:83–88Google Scholar
  9. Dykhuizen D, Hartl D (1981) Evolution of competitive ability in Escherichia coli. Evolution 35:581–594Google Scholar
  10. Germain P, Toukourou F, Donaduzzi L (1986) Ethanol production by anaerobic thermophilic bacteria: regulation of lactate dehydrogenase activity in Clostridium thermhydrosulfuricum. Appl Microbiol Biotechnol 24:300–305Google Scholar
  11. Ghazvinizadeh H, Turtura GC, Zambonelli C (1972) The fermentation of L-rhamnose by clostridia. Ann Microbiol Enzimol 22:155–158Google Scholar
  12. Gong C-S, McCracken LD, Tsao GT (1981) Direct fermentation of D-xylose to ethanol by a xylose-fermentating yeast mutant, Candida sp. XF217. Biotechnol Lett 3:245–250Google Scholar
  13. Kannan V, Mutharasan R (1985) Ethanol fermentation characteristics of Thermoanaerobacter ethanolicus. Enzyme Microbiol Technol 7:87–89Google Scholar
  14. Kersters K, De Ley J (1980) Classification and identification of bacteria by electrophoresis of their proteins. In: Goodfellow M, Board RG (eds) Microbiological classification and identification. Academic Press, TorontoGoogle Scholar
  15. Lacis LS, Lawford HG (1985) Thermoanaerobacter ethanolicus in a comparison of the growth efficiencies of thermophilic and mesophilic anaerobes. J Bacteriol 163:1275–1278Google Scholar
  16. Ljungdahl LG, Carreira LH (1983) High ethanol producing derivatives of Thermoanaerobacter ethanolicus. US Patent 4,385,117Google Scholar
  17. Lynd LR, Grethlein HE (1984) IHOSR/extractive distillation for ethanol separation. Chem Eng Prog 80:59–62Google Scholar
  18. Margaritis A, Bajpai P (1982) Direct fermentation of D-xylose to ethanol by Kluyveromyces marxianus strains. Appl Environ Microbiol 44:1039–1041Google Scholar
  19. Mian FA, Fencl Z, Prokoi A, Mohagheghi A, Fazeli A (1974) Effect of growth rate on the glucose metabolism of yeast grown in continuous culture. Radiorespirometric studies. Folia Microbiol 19:191–198Google Scholar
  20. Mistry FR, Cooney CL (1985) Ethanol production by C. thermosaccharolyticum in a continuous culture cell-recycle system. Preprint, Am Chem Soc Ann Meeting, Chicago, USAGoogle Scholar
  21. Muetze B, Wandrey C (1983) Continuous fermentation of xylose with Pachysolen tannophilus. Biotechnol Lett 5:633–638Google Scholar
  22. Neijssel OM, Tempest DW (1976) Bioenergetic aspects of aerobic growth of Klebsiella aerogenes NCTC 418 in carbon-limited and carbon-sufficient chemostat culture. Arch Microbiol 107:215–221Google Scholar
  23. Nigam JN, Ireland RS, Margaritis A, Lachance MA (1985a) Isolation and screening of yeasts that ferment D-xylose directly to ethanol. Appl Environ Microbiol 50:1486–1489Google Scholar
  24. Nigam JN, Margaritis A, Lachance MA (1985) Aerobic fermentation of D-xylose to ethanol by Clavispora sp. Appl Environ Microbiol 50:763–766Google Scholar
  25. Patel GB, MacKenzie R, Agnew BJ (1986) Fermentation of xylose and hemicellulose hydrolysates by an ethanol-adapted culture of Bacteroides polypragmatus. Arch Microbiol 146:68–73Google Scholar
  26. Payton MA (1984) Production of ethanol by thermophilic bacteria. Trends Biotechnol 2:153–158Google Scholar
  27. Pirt SJ (1975) Principles of microbe and cell cultivation. John Wiley and Sons, New YorkGoogle Scholar
  28. Powell EO (1958) Criteria for the growth of contaminants and mutants in continuous culture. J Gen Microbiol 18:259–268Google Scholar
  29. Preez JC du, Bosch M, Prior BA (1986) Xylose fermentation by Candida shehatae and Pichia stipitis: effects of pH, temperature and substrate concentration. Enzyme Microb Technol 8:360–364Google Scholar
  30. Sayed IA, Kenny GE (1980) Comparison of the proteins and polypeptides of the eight serotypes of Ureaplasma urealyticum by isoelectric focusing and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Int J Syst Bacteriol 30:33–41Google Scholar
  31. Slininger PJ, Bothast RJ, Okos MR, Ladisch MR (1985) Comparative evaluation of ethanol production by xylose-fermenting yeast presented high xylose concentrations. Biotechnol Lett 7:431–436Google Scholar
  32. Sonnleitner B (1983) Biotechnology of thermophilic bacteriagrowth, products, and application. In: Fiechter A (ed) Advances in biochemical engineering and biotechnology, vol 28. Springer, New YorkGoogle Scholar
  33. Sonnleitner B, Fiechter A (1983) Advantages of using thermophiles in biotechnological processes: expectations and reality. Trends Biotechnol 1:74–80Google Scholar
  34. Sonnleitner B, Fiechter A, Giovannini F (1984) Growth of Thermoanaerobium brockii in batch and continuous culture at supraoptimal temperatures. Appl Microbiol Biotechnol 19: 326–334Google Scholar
  35. Thomas TD, Ellwood DC, Longyear VMC (1979) Change from homo- to heterolactic fermentation by Streptococcus lactis resulting from glucose limitation in anaerobic chemostat cultures. J Bacteriol 138:109–117Google Scholar
  36. Tran-Din K, Gottschalk G (1985) Formation of D(-)-1,2-propanediol and D(-)-lactate from glucose by Clostridium sphenoides under phosphate limitation. Arch Microbiol 142:87–92Google Scholar
  37. Turner KW, Robertson AM (1979) Xylose, arabinose, and rhamnose fermentation by Bacteroides ruminocola. Appl Environ Microbiol 38:7–12Google Scholar
  38. Veldkamp H, Jannasch HW (1972) Mixed culture studies with the chemostat. J Appl Chem Biotechnol 22:105–123Google Scholar
  39. Wang DIC, Fang I-Y (1980) Ethanol production by a thermophilic and anaerobic bacterium using xylose. Am Chem Soc Div Pet Chem 25:639–649Google Scholar
  40. Wang DIC, Cooney CI, Demain AL, Dunnill P, Humphrey AE, Lilly MD (1979) Fermentation and enzyme technology. John Wiley & Sons, New York Chichester Brisbane TorontoGoogle Scholar
  41. Weimer PJ (1984) Fermentation of 6-deoxyhexoses by Bacillus macerans. Appl Environ Microbiol 47:263–267Google Scholar
  42. Weimer PJ (1985) Thermophilic anaerobic fermentation of hemicellulose and hemicellulose-derived aldose sugars by Thermoanaerobacter strain B6A. Arch Microbiol 143:130–136Google Scholar
  43. Wiegel J (1980) Formation of ethanol by bacteria. A pledge for the use of extreme thermophilic anaerobic bacteria in industrial ethanol fermentation processes. Experientia 36:1434–1446Google Scholar
  44. Wiegel J, Ljungdahl LG (1981) Thermoanaerobacter ethanolicus gen. nov., spec. nov., a new, extreme thermophilic, anaerobic bacterium. Arch Microbiol 128:343–348Google Scholar
  45. Wiegel J, Carreira LH, Mothershed CP, Puls J (1983) Production of ethanol from biopolymers by anaerobic, thermophilic and extreme thermophilic bacteria. II. Thermoanaerobacter ethanolicus JW200 and its mutants in batch cultures and resting cell experiments. Biotechnol Bioeng Symp 13:193–205Google Scholar
  46. Woods MA, Millis NF (1985) Effect of slow feeding of xylose on ethanol yield by Pachysolen tannophilus. Biotechnol Lett 7:679–682Google Scholar
  47. Zeikus JG (1979) Thermophilic bacteria: ecology, physiology and technology. Enzyme Microb Technol 1:243–252Google Scholar

Copyright information

© Springer-Verlag 1988

Authors and Affiliations

  • Lynda S. Lacis
    • 1
  • Hugh G. Lawford
    • 1
  1. 1.University of TorontoTorontoCanada

Personalised recommendations