Butanol Dehydrogenase Research Articles

Enzymatic investigations on butanol dehydrogenase and butyraldehyde dehydrogenase in extracts of Clostridium acetobutylicum

Reliable assay systems were developed for the detection and quantitation of butanol dehydrogenase and butyraldehyde dehydrogenase in extracts of Clostridium acetobutylicum. Butanol dehydrogenase was NADPH-dependent. The enzyme could be sparated by ultracentrifugation from a NADH-specific enzyme which probably represents the ethanol dehydrogenase but which also reacted with butyraldehyde to form butanol. Butyraldehyde dehydrogenase proved to be NADH-specific. All enzymes were induced shortly before butanol formation began. Specific activities decreased at the end of the fermentation process. An explanation for contradictory data in the literature is proposed.

Level of enzymes involved in acetate, butyrate, acetone and butanol formation by Clostridium acetobutylicum

Clostridium acetobutylicum cells were collected from chemostats which were run at pH 4.3 or 6.0 and which produced either acetone-butanol or acetate-butyrate; they were used to determine the level of enzymes involved either in solvent or in acid formation. The highest activity of phosphotransacetylase, phosphotransbutyrylase, acetate kinase, and butyrate kinase was found in cells which carried out an acetate-butyrate fermentation; these enzymes were present in solvent-producing cells at a level of about 10–50% as compared to acid-producing cells. Hydrogenase activity was detectable in approximately the same amounts in both cell types; however, in solvent-producing cells it was only measurable following a lag-period. Butyraldehyde and butanol dehydrogenases were found in small amounts exclusively in solvent-producing cells. It was demonstrated that the formation of acetone was initiated by the action of a coenzyme A-transferase which transferred coenzyme A from acetoacetyl-CoA to either acetate or butyrate. This coenzyme A-transferase as well as acetoacetate decarboxylase were hardly detectable in acid-producing cells, but reached high levels in solvent producing cells. Similar changes of the activity of the enzymes mentioned were observed when a batch culture was shifted from acid to solvent formation.

Acidic Conditions Are Not Obligatory for Onset of Butanol Formation by Clostridium beijerinckii (Synonym, C. butylicum )

Factors that may initiate the metabolic transition for butanol production were investigated in batch cultures of Clostridium beijerinckii (synonym, Clostridium butylicum) VPI 13436. Cultures maintained at pH 6.8 produced nearly as much butanol as those incubated without pH control, indicating that neither a change in the culture pH nor acid conditions per se are always required to initiate solvent formation. Acetate and butyrate levels at the onset of butanol production were dependent on the pH at which the cultures were maintained. Cultures maintained at pH 6.8 could be accelerated into solvent production by artificially lowering the pH to 5.0 or by the addition of acetate plus butyrate without a pH change (but neither acid alone was effective). Solvent production was associated with slower rates of growth and general metabolism, and it did not show a requirement for mature spore formation. We speculate that a slowdown in metabolism, which may be brought about by several conditions, is mechanistically related to the onset of butanol production. Extracts of solvent-producing cells contained acetoacetate decarboxylase activity as well as higher NADP-linked butanol dehydrogenase and lower hydrogenase activities than extracts of acid-producing cells. Solvent production did not appear to involve an enhanced ability to catalyze H(2) oxidation.