ABSTRACT
Repeated fed-batch fermentation of glucose by Clostridium tyrobutyricum immobilized in a fibrous bed bioreactor (FBB) was successfully employed to produce butyric acid at a high final concentration as well as to adapt a butyric-acid-tolerant strain. At the end of the eighth fed-batch fermentation, the butyric acid concentration reached 86.9 ± 2.17 g/L, which to our knowledge is the highest butyric acid concentration ever produced in the traditional fermentation process. To understand the mechanism and factors contributing to the improved butyric acid production and enhanced acid tolerance, adapted strains were harvested from the FBB and characterized for their physiological properties, including specific growth rate, acid-forming enzymes, intracellular pH, membrane-bound ATPase and cell morphology. Compared with the original culture used to seed the bioreactor, the adapted culture showed significantly reduced inhibition effects of butyric acid on specific growth rate, cellular activities of butyric-acid-forming enzyme phosphotransbutyrylase (PTB) and ATPase, together with elevated intracellular pH, and elongated rod morphology.
Subject(s)
Bioreactors/microbiology , Butyric Acid/metabolism , Butyric Acid/toxicity , Cells, Immobilized/drug effects , Cells, Immobilized/metabolism , Clostridium tyrobutyricum/drug effects , Clostridium tyrobutyricum/metabolism , Adenosine Triphosphatases/metabolism , Anti-Bacterial Agents/metabolism , Anti-Bacterial Agents/toxicity , Cell Membrane/enzymology , Clostridium tyrobutyricum/cytology , Clostridium tyrobutyricum/enzymology , Cytosol/chemistry , Fermentation , Glucose/metabolism , Hydrogen-Ion Concentration , Microscopy , Phosphate Acetyltransferase/metabolismABSTRACT
The effects of the hydraulic retention time (HRT=8, 10, 12 or 16.7 h) and glucose concentration (30, 40 or 50 g/L) on the production of hydrogen and butyrate by an immobilized Clostridium tyrobutyricum culture, grown under continuous culturing conditions, were evaluated. With 30 g/L glucose, the higher HRTs tested led to greater butyrate concentrations in the culture, i.e., 9.3 g/L versus 12.9 g/L with HRTs of 8 h and 16.7 h, respectively. In contrast, higher biogas and hydrogen production rates were generally seen when the HRT was lower. Experiments with different glucose concentrations saw a significant amount of glucose washed out when 50 g/L was used, the highest being 22.7 g/L when the HRT was 16.7 h. This study found the best conditions for the continuous production of hydrogen and butyric acid by C. tyrobutyricum to be with an HRT of 12 h and a glucose concentration of 50 g/L, respectively.
