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The initial growth of Clostridium acetobutyilcum was not inhibited by I atm of H_2 while H_2 reduced glucose consumption in a solventogenic culture of a phosphate limited 2-stage chemostat. Under 1 atm of H_2, a solventogenic culture consumed hydrogen, but an acidogenic culture produced hydrogen. H_2 consumption by the solventogenic culture was enhanced by the addition of 5mM neutral red, an artificial electron carrier with a redox potential of -325 mV. Hydrogenase activity, measured in both directions of production and consumption, showed that activity coupled with methyl viologen is higher in an acidogenic culture than in a solventogenic culture, and that the two cultures have similar activities for methylene blue reduction. The solventogenic culture showed a higher activity coupled with neutral red than the acidogenic culture. From these results, it is hypothesized that hydrogen producing hydrogenase activity is high during the acidogenic phase, and decreases as solventogenesis starts, and that the solventogenic culture produces a second hydrogenase which uses an electron carrier other than ferredoxin. This hypothesis was supported by the fact that enzyme activities involved in electron flow can be coupled to neutral red, indepedent of ferredoxin, and that neutral red addition to the fermentation system increased butanol yield, with a decrease in production of less reduced fermentation products, and H_2.
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