Abstract
A periplasmic thiosulfate dehydrogenase (EC 1.8.2.2) was purified to homogeneity from the neutrophilic, obligately chemolithoautotrophicThiobacillus sp. W5. A five-step procedure resulted in an approximately 2,300-fold purification. The purified protein had a molecular mass of 120±3 kDa, as determined by gel filtration. It is probably a tetramer containing two different subunits with molecular masses of 33±1 kDa and 27±0.5 kDa, as determined by SDS-PAGE. UV/visible spectroscopy revealed that the enzyme contained haemc; haem staining showed that both subunits contained haemc. A haemc content of 4 mol per mol of enzyme was calculated using the pyridine haemochrome test. The pH optimum of the enzyme was 5.5 At pH 7.5, the Km and Vmax were 120±10 μM and 1,160±30 U mg-1, respectively. The absence of 2-heptyl-4-hydroquinoline-N-oxide (HQNO) inhibition for the oxidation of thiosulfate by whole cells suggested that the electrons enter the respiratory chain at the level of cytochromec. Comparison with thiosulfate dehydrogenases from otherThiobacillus species showed that the enzyme was structurally similar to the thiosulfate dehydrogenase of the acidophilic, facultatively chemolithoautotrophicThiobacillus acidophilus, but not to the thiosulfate dehydrogenases published for the obligately chemolithoautotrophicThiobacillus tepidarius andThiobacillus thioparus.
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Abbreviations
- BV :
-
Benzyl viologen
- DCPIP :
-
2,6-Dichloroindophenol
- HQNO :
-
2-Heptyl-4-hydroquinoline-N-oxide
- NEM :
-
N-ethylmaleimide
- PES :
-
Phenazine ethosulfate
- PMS :
-
Phenazine methosulfate
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Visser, J.M., de Jong, G.A.H., Robertson, L.A. et al. Purification and characterization of a periplasmic thiosulfate dehydrogenase from the obligately autotrophicThiobacillus sp. W5. Arch. Microbiol. 166, 372–378 (1996). https://doi.org/10.1007/BF01682982
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DOI: https://doi.org/10.1007/BF01682982