Metabolic control of Clostridium thermocellum via inhibition of hydrogenase activity and the glucose transport rate

Hsin Fen Li, Barbara L. Knutson, Sue E. Nokes, Bert C. Lynn, Michael D. Flythe

Research output: Contribution to journalArticlepeer-review

14 Scopus citations


Clostridium thermocellum has the ability to catabolize cellulosic biomass into ethanol, but acetic acid, lactic acid, carbon dioxide, and hydrogen gas (H 2) are also produced. The effect of hydrogenase inhibitors (H 2, carbon monoxide (CO), and methyl viologen) on product selectivity was investigated. The anticipated effect of these hydrogenase inhibitors was to decrease acetate production. However, shifts to ethanol and lactate production are also observed as a function of cultivation conditions. When the sparge gas of cellobiose-limited chemostat cultures was switched from N 2 to H 2, acetate declined, and ethanol production increased 350%. In resting cell suspensions, lactate increased when H 2 or CO was the inhibitor or when the cells were held at elevated hyperbaric pressure (6.8 atm). In contrast, methyl-viologen-treated resting cells produced twice as much ethanol as the other treatments. The relationship of chemostat physiology to methyl viologen inhibition was revealed by glucose transport experiments, in which methyl viologen decreased the rate of glucose transport by 90%. C. thermocellum produces NAD + from NADH by H 2, lactate, and ethanol production. When the hydrogenases were inhibited, the latter two products increased. However, excess substrate availability causes fructose 1,6-diphosphate, the glycolytic intermediate that triggers lactate production, to increase. Compensatory ethanol production was observed when the chemostat fluid dilution rate or methyl viologen decreased substrate transport. This research highlights the complex effects of high concentrations of dissolved gases in fermentation, which are increasingly envisioned in microbial applications of H 2 production for the conversion of synthetic gases to chemicals.

Original languageEnglish
Pages (from-to)1777-1784
Number of pages8
JournalApplied Microbiology and Biotechnology
Issue number4
StatePublished - Feb 2012

Bibliographical note

Funding Information:
Acknowledgements The authors gratefully acknowledge the financial support of the Southeastern Sun Grant Center, administered by the University of Tennessee, grant number DTOS59-07-G-0050. MF was supported by the Agricultural Research Service, USDA. The authors would like to thank Gloria Gellin and Jerry Vice for the technical assistance.


  • Cellulose
  • Cellulosic biomass conversion
  • Chemostat
  • Consolidated bioprocessing
  • Continuous culture
  • Metabolic engineering
  • Paraquat
  • Product selectivity

ASJC Scopus subject areas

  • Biotechnology
  • Applied Microbiology and Biotechnology


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