Energetics of mixotrophic and autotrophic C1-metabolism by Thiobacillus acidophilus

Pronk, Jack T.; Bruijn, Peter de; Dijken, Johannes P. van; Bos, P.; Kuenen, J.G.

doi:10.1007/bf00248839

Cited by 9 publications

(5 citation statements)

References 30 publications

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“…The strain expressed 0.41 units of FDH activity per ml of the culture broth, and was identied as Thiobacillus sp., the FDH of which has been reported previously. [35][36][37][38][39] Puriˆcation and properties of FDH The FDH was puriˆed about 26-fold ( Table 2). The puriˆed enzyme was a single protein band of 44 kDa on SDS-PAGE.…”

Section: Resultsmentioning

confidence: 99%

Purification and Characterization of an α-Haloketone-resistant Formate Dehydrogenase fromThiobacillussp. Strain KNK65MA, and Cloning of the Gene

Nanba

Takaoka

Hasegawa

2003

Bioscience, Biotechnology, and Biochemistry

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Thiobacillus sp. strain KNK65MA, which produced an NAD-dependent formate dehydrogenase (FDH) highly resistant to alpha-haloketones, was newly isolated, i.e., the enzyme showed no loss of activity after a 5-h incubation with alpha-haloketones, such as ethyl 4-chloro-3-oxobutanoate. The enzyme was also resistant to SH reagents. The enzyme, purified to homogeneity, was a dimer composed of identical subunits. The specific activity was 7.6 u/mg, and the apparent Km values for formate and NAD+ were 1.6 and 0.048 mM, respectively. The cloned gene of FDH contained one open reading frame (ORF) of 1206 base pairs, predicted to encode a polypeptide of 401 amino acids, with a calculated molecular weight of 44,021; this gene was highly expressed in E. coli cells. The deduced amino acid sequence of this FDH had high identity to other bacterial FDHs.

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Section: Resultsmentioning

confidence: 99%

Purification and Characterization of an α-Haloketone-resistant Formate Dehydrogenase fromThiobacillussp. Strain KNK65MA, and Cloning of the Gene

Nanba

Takaoka

Hasegawa

2003

Bioscience, Biotechnology, and Biochemistry

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“…However, in contrast to the M. gastri formate dehydrogenase, the T. ferrooxidans enzyme appeared to be located in the soluble fraction of cell extracts. The T. ferrooxidans enzyme activity differs markedly from the T. acidophilus formate dehydrogenase activity, which can use both NAD and artificial electron acceptors (23).…”

Section: Discussionmentioning

confidence: 98%

“…Therefore, the high substrate concentrations used in such experiments are likely to inhibit growth because of substrate toxicity. Earlier studies in our laboratory revealed that the facultatively autotrophic acidophile T. acidophilus could utilize formate as an energy source when this substrate was provided at low concentrations (23). It * Corresponding author.…”

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confidence: 99%

Growth of Thiobacillus ferrooxidans on Formic Acid

Pronk

Meijer

Hazeu

et al. 1991

Appl Environ Microbiol

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A variety of acidophilic microorganisms were shown to be capable of oxidizing formate. These included Thiobacillusferrooxidans ATCC 21834, which, however, could not grow on formate in normal batch cultures. However, the organism could be grown on formate when the substrate supply was growth limiting, e.g., in formate-limited chemostat cultures. The cell densities achieved by the use of the latter cultivation method were higher than cell densities reported for growth of T. ferrooxidans on ferrous iron or reduced sulfur compounds. Inhibition of formate oxidation by cell suspensions, but not cell extracts, of formate-grown T. ferrooxiddns occurred at formate concentrations above 100 ,iM. This observation explains the inability of the organism to grow on formate in batch cultures. Cells grown in formate-limited chemostat cultures retained the ability to oxidize ferrous iron at high rates. Ribulose 1,5-bisphosphate carboxylase activities in cell extracts indicated that T. ferrooxidans employs the Calvin cycle for carbon assimilation during growth on formate. Oxidation of formate by cell extracts was NAD(P) independent.

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“…Carbon sources supporting heterotrophic growth include a number of monosaccharides, tricarboxylic-acid-cycle intermediates and some amino acids (Guay and Silver, 1975;Pronk et al, 1990a). Growth substrates for autotrophic growth include formate (Pronk et al, 1990d), elemental sulphur (Guay and Silver, 1975), tetrathionate (Norris et al, 1986), thiosulphate (S,O:-) and trithionate (Mason et al, 1987).…”

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confidence: 99%

Purification and paritial characterization of a thermostable trithionate hydrolase from the acidophilic sulphur oxidizer Thiobacillus acidophilus

Meulenberg¹,

Pronk²,

Frank³

et al. 1992

European Journal of Biochemistry

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Ccll-free extracts of Thiobacillus acidophilus catalysed the quantitative conversion of trithionate (S,Oz -) to thiosulphate and sulphate. A continuous assay for quantification of experimental results was based on the difference in absorbance between trithionate and thiosulphate at 220 nm.Trithionate hydrolase was purified to near homogeneity from cell-free extracts of T . acidophilus. The molecular masses of the native enzyme and the subunit were 99 kDa (gel filtration) and 34 kDa (SDS/PAGE). The purified enzyme has a pH optimum of 3.5-4.5 and a temperature optimum of 70 "C. Enzyme activity was stimulated by sulphate. The stimulation of the enzyme activity by sulphate was half maxima! at a concentration of 0.23 M. The K, for trithonate is 70 pM at 30°C and 270 pM at 70°C. Enzyme activity was lost after 36 days at O"C, 27 days at 70°C; but after 97 days at 30"C, 40% of the initial activity was still present. The enzyme activity was inhibited by mercury chloride, .hi-ethylmaleimidc, thiosulphate and tetrathionate. Tetrathionate S402-was not hydrolysed by trithionate hydrolase.

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Energetics of mixotrophic and autotrophic C1-metabolism by Thiobacillus acidophilus

Cited by 9 publications

References 30 publications

Purification and Characterization of an α-Haloketone-resistant Formate Dehydrogenase fromThiobacillussp. Strain KNK65MA, and Cloning of the Gene

Purification and Characterization of an α-Haloketone-resistant Formate Dehydrogenase fromThiobacillussp. Strain KNK65MA, and Cloning of the Gene

Growth of Thiobacillus ferrooxidans on Formic Acid

Purification and paritial characterization of a thermostable trithionate hydrolase from the acidophilic sulphur oxidizer Thiobacillus acidophilus

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