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Pseudomonas aeruginosa ATCC 17933 is able to oxidize ethanol to acetate under aerobic conditions. The P. aeruginosa acetyl-CoA synthetase (ACS) gene acsA was previously identified, and the ACS enzyme described to be required for growth on ethanol as the sole source of carbon and energy. Here, we investigated the transcriptional regulation of the acsA gene using an acsA::lacZ fusion. Transcription of acsA was regulated by the carbon source, and expression was maximal on ethanol, acetate and propionate. In addition, the induction depended on the response regulator ErdR, which also regulates hierarchically arranged genes for ethanol oxidation. Transcription of the acsA gene was repressed by addition of succinate to an ethanol-containing medium. This repression required Crc, the product of the catabolite repression control gene crc.

The anaerobic hyperthermophilic archaea Desulfurococcus amylolyticus, Hyperthermus butylicus, Thermococcus celer, Pyrococcus woesei, the hyperthermophilic bacteria Thermotoga maritima and Clostridium thermohydrosulfuricum and the aerobic mesophilic archaeon Halobacterium saccharovorum were grown either on complex media, on sugars or on pyruvate as carbon and energy sources. During growth acetate was formed as fermentation product by all organisms. The enzymes involved in acetyl-CoA formation from pyruvate and in acetate formation from acetyl-CoA were investigated: These data indicate that acetyl-CoA synthetase (ADP forming) represents a typical archaeal property rather than an enzyme specific for hyperthermophiles. It is proposed that in all acetate forming archaea the formation of acetate and of ATP from acetyl-CoA, ADP and Pi are catalyzed by acetyl-CoA synthetase (ADP forming), whereas in all acetate forming (eu)bacteria these reactions are catalyzed by two enzymes, phosphate acetyltransferase and acetate kinase.

The hyperthermophilic, sulfate-reducing archaeon Archaeoglobus fulgidus strain 7324, rather than the type strain VC16, was found to grow on starch and sulfate as energy and carbon source. Fermentation products and enzyme activities were determined in starch-grown cells and compared to those of cells grown on lactate and sulfate. During exponential growth on starch, 1 mol of glucose-equivalent was incompletely oxidized with sulfate to approximately 2 mol acetate, 2 mol CO2 and 1 mol H2S. Starch-grown cells did not contain measurable amounts of the deazaflavin factor F420 (

All the enzymes required for the oxidation of acetate to CO2 via the citric acid cycle were found in Desulfobacter postgatei. To obtain in vivo evidence for the operation of this cycle, the sulfate reducing bacterium was grown on [14C]acetate in the presence of a large pool of 12CO2 and the incorporation of 14C into glutamate (≙ 2-oxoglutarate), aspartate (≙ oxaloacetate), and alanine (≙ pyruvate) was studied. The labelling data were found to be consistent with (i) the oxidation of acetate to CO2 via the reactions of the citric acid cycle, (ii) the synthesis of citrate via a citrate (si)-synthase, and (iii) the anaplerotic synthesis of oxaloacetate from acetate and 2 CO2 via pyruvate as intermediate.

Acetate uptake by strains of Synechococcus and Aphanocapsa in short experiments required light, and was strongly inhibited by m-dichlorocarbonyl cyanide phenylhydrazone and dichlorophenyl dimethyl urea. Acetate carbon was distributed in amino acids and in the acyl portion of lipids in the same way as during growth experiments when CO2 was available, but the reduced incorporation in the absence of CO2 was primarily into the lipid fraction. An apparent K m for uptake by Synechococcus and for Aphanocapsa 6308 of 20 and 180 μM at pH 7.4 was obtained; corresponding V max values were 6 and 11 nmol x min-1 x mg protein-1. Uptake with Synechococcus was affected by pH, with affinity decreased and maximal rate increase with rising pH. Acetate uptake was not affected by propionate or butyrate when both were added at the same time, but a light and concentration dependent inhibition developed if suspensions were preincubated with propionate. Acetate carbon moved rapidly into acid insoluble material, but after 10–15 s 75% or more of the recovered intracellular counts were in acetyl CoA. Counts in this compound were reduced by preincubation with propionate. Kinetic measurements of acetyl CoA synthetase in fractionated cell extracts gave values for K m of about 50 μM for acetate, 5 mM for propionate, 100 μM for CoA and 0.38 mM for ATP. The internal pool of free CoA was measured to be about 20 μM, and was reduced by preincubation with propionate. This suggests that the activity of CoA-mediated reactions may be regulated by the availability of this cofactor.

Methanobacterium thermoautotrophicum growing on H2 plus CO2 as sole carbon and energy source was found to contain acetate thiokinase (Acetyl CoA synthetase; EC 6.2.1.1); Acetate + ATP + CoA leads to Acetyl CoA + AMP + PPi. The apparent Km value for acetate was 40 microM. Acetate kinase (EC 2.7.2.1) and phosphotransacetylase (EC 2.3.1.8) could not be detected. The specific activity of acetate thiokinase was high in cells grown with limited H2 and CO2 supply (approximately 100 nmol/min . mg protein), it was low in exponentially grown cells (2 nmol/min . mg protein). This corresponded with the finding that cells growing linearly in the presence of acetate assimilated the monocarboxylic acid in high amounts (greater than 10% of the cell carbon was derived from acetate), whereas exponentially growing cells did not (less than 1% of cell carbon was derived from acetate). These latter observations indicated that acetate thiokinase and free acetate are not involved in autotrophic CO2 fixation in M. thermoautotrophicum. The presence and some kinetic properties of succinate thiokinase (EC 6.2.1.5), adenylate kinase (EC 2.7.4.3), and inorganic pyrophosphatase (EC 3.6.1.1) are also described.