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Recombinant phosphoribosyl pyrophosphate synthetases from Thermus thermophilus HB27: Isolation and properties

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Abstract

Two genes of T. thermophilus HB27, TT_C1184 and TT_C1274, encoding proteins with phosphoribosyl pyrophosphate synthetase activity were cloned in an expressing vector pET23d+. Escherichia coli strains overproducing two relevant proteins in soluble forms were obtained. The methods of isolation of thermophilic phosphoribosyl pyrophosphate synthetases Tth PRPPS1 and Tth PRPPS2 were developed. The activities of these enzymes were determined as a function of concentration of metal ions, inorganic phosphate, and temperature. The kinetic parameters for basic natural substrates were calculated; the substrate specificity for different carbohydrate 5-phosphates of D-series was studied. It was shown that the two proteins differ significantly in these characteristics. According to the results and comparison of amino acid sequences of new proteins with those of other phosphoribosyl pyrophosphate synthetases, both enzymes belong to class I PRPPS.

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Authors and Affiliations

  1. Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, ul. Miklukho-Maklaya 16/10, Moscow, 117997, Russia

    R. S. Esipov, Yu. A. Abramchik, I. V. Fateev, T. I. Muravyova, K. G. Artemova, I. D. Konstantinova & A. I. Miroshnikov

  2. Shubnikov Institute of Crystallography, Russian Academy of Sciences, Leninski pr. 59, Moscow, 119333, Russia

    I. P. Kuranova

  3. National Research Center “Kurchatov Institute”, pl. akad. Kurchatova 1, Moscow, 123182, Russia

    I. P. Kuranova

Authors
  1. R. S. Esipov
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  2. Yu. A. Abramchik
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  3. I. V. Fateev
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  4. T. I. Muravyova
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  5. K. G. Artemova
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  6. I. D. Konstantinova
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  7. I. P. Kuranova
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  8. A. I. Miroshnikov
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Correspondence to R. S. Esipov.

Additional information

Original Russian Text © R.S. Esipov, Yu.A. Abramchik, I.V. Fateev, T.I. Muravyova, K.G. Artemova, I.D. Konstantinova, I.P. Kuranova, A.I. Miroshnikov, 2016, published in Bioorganicheskaya Khimiya, 2016, Vol. 42, No. 5, pp. 567–577.

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Esipov, R.S., Abramchik, Y.A., Fateev, I.V. et al. Recombinant phosphoribosyl pyrophosphate synthetases from Thermus thermophilus HB27: Isolation and properties. Russ J Bioorg Chem 42, 512–521 (2016). https://doi.org/10.1134/S1068162016040075

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  • DOI: https://doi.org/10.1134/S1068162016040075

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