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Exogenous H2/CO2 and glucose were consumed simultaneously by Butyribacterium methylotrophicum when grown under glucose-limited conditions. CO2 reduction to acetate was coupled to H2 consumption. The addition of either H2 or CO2 to glucose batch fermentation resulted in an increase in cell density, hydrogenase (H2-consuming and -producing) activities and fatty acid production by B. methylotrophicum as compared to when N2 was the feed gas. Hydrogenase activities appeared to be tightly regulated and were produced at higher rates during the exponential phase when CO2 was the feed gas as compared to H2 or N2. The increase in H2-consuming activity and decrease in H2-producing activity was correlated with an increase in butyrate synthesis. H2-consuming and ferredoxin (Fd)–NAD reductase activities increased while H2-producing and NADH–Fd reductase activities decreased in cells grown at pH 5.5 compared to those at pH 7.0. The molar ratio of butyrate/acetate was shifted from 0.35 at pH 7.0 to 1.22 at pH 5.5. The addition of exogenous H2 did not decrease the butyrate/acetate ratio at pH 7.0 nor at pH 5.5. The results indicated that growth pH values regulated both hydrogenase and Fd–NAD oxidoreductase activities such that, at acid pH, more intermediary electron flow was directed towards butyrate synthesis than H2 production.