1995
DOI: 10.1128/jb.177.10.2878-2886.1995
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Abstract: Acetyl coenzyme A synthetase (Acs) activates acetate to acetyl coenzyme A through an acetyladenylate intermediate; two other enzymes, acetate kinase (Ack) and phosphotransacetylase (Pta), activate acetate through an acetyl phosphate intermediate. We subcloned acs, the Escherichia coli open reading frame purported to encode Acs (F. R. Blattner, V. Burland, G. Plunkett III, H. J. Sofia, and D. L. Daniels, Nucleic Acids Res. 21:5408-5417, 1993). We constructed a mutant allele, ⌬acs::Km, with the central 0.72-kb B… Show more

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Cited by 217 publications
(198 citation statements)
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“…Though lactate and formate are less toxic to the cells, they are still disadvantageous for cell growth. The enzyme has a strong affinity for acetate (Km of 200µM), which allows it to function at low acetate concentrations, but it is inhibited by glucose [66,67]. On the other hand, the reversible ackA-pta pathway can assimilate acetate only at high acetate concentrations [15].…”
Section: Pep Carboxylase (Ppc) and Pep Carboxykinase (Pck)mentioning
confidence: 99%
“…Though lactate and formate are less toxic to the cells, they are still disadvantageous for cell growth. The enzyme has a strong affinity for acetate (Km of 200µM), which allows it to function at low acetate concentrations, but it is inhibited by glucose [66,67]. On the other hand, the reversible ackA-pta pathway can assimilate acetate only at high acetate concentrations [15].…”
Section: Pep Carboxylase (Ppc) and Pep Carboxykinase (Pck)mentioning
confidence: 99%
“…When the E. coli wild-type cells or the cells deficient in Acs (acetylCoA synthetase) (strain AJW1781) were inoculated into TB medium (37) and aerated at 37°C, the wild-type cells and Acs-deficient cells grew at similar rates and excreted similar amounts of acetate before the mid-exponential phase. Although both types of cells continued to grow at similar rates, the extracellular acetate concentration differed.…”
Section: Cloning and Nucleotide Sequence Of The 3ј Downstreammentioning
confidence: 99%
“…Although both types of cells continued to grow at similar rates, the extracellular acetate concentration differed. Whereas the concentration of extracellular acetate in the wild-type cells decreased to a minimum of Ͻ0.1 mM, that in the Acs-deficient cells did not decrease at all (37). We transformed AJW1781 with plasmid pUC-acsA, which causes the expression of AcsA.…”
Section: Cloning and Nucleotide Sequence Of The 3ј Downstreammentioning
confidence: 99%
“…Attempts to obtain complementation using the C. themtocellurn libraries have not however been successful to date. Cloning via complementation was complicated by occasional mutant reversion, and by leaky growth of the E. coli mutants on minimal acetate plates due to an acetate assimilation pathway mediated by acetyl-coA synthetase (96,204).…”
mentioning
confidence: 99%
“…Attempts to obtain complementation using the C. themtocellurn libraries have not however been successful to date. Cloning via complementation was complicated by occasional mutant reversion, and by leaky growth of the E. coli mutants on minimal acetate plates due to an acetate assimilation pathway mediated by acetyl-coA synthetase (96,204).In light of the difficulties encountered with cloning by complementation, we investigated a cloning strategy using the Polymerase Chain Reaction (PCR) instead. Using available sequence information, PCR primers were synthesized designed to amplify fragments of genes coding for lactic dehydrogenase (ldh), acetate kinase (ack), phosphotransacetylase @fa), and hydrogenase (hyd).…”
mentioning
confidence: 99%