Dehydrogenase (DH) activity associated with bio-electrochemical behavior was analyzed for the first time to understand the redox reactions involved in fermentative hydrogen (H(2)) production process in concurrence with proton (H(+)) shuttling and electron (e(-)) discharge (ED) pattern. DH facilitates the availability of H(+) through redox reactions to make H(2). We have designed a comprehensive experimental study to evaluate the DH activity (H(+) shuttling) and ED to understand the biochemical process with the function of pH (5, 6, 7 and 8) and metabolic microenvironment [anaerobic, anoxic and aerobic (control)]. DH activity was observed to be higher during anaerobic operation suggesting the higher availability of H(+) and e(-) due to the inter-conversion of metabolites and the same was reflected in the voltammetry analysis. Higher H(2) production under anaerobic operation corroborated well with these findings. The DH activity associated with H(+) shuttling and ED was also correlated with the substrate degradation pattern.
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