Recombinant phosphoribosyl pyrophosphate synthetases from Thermus thermophilus HB27: Isolation and properties
- Authors: Esipov R.S.1, Abramchik Y.A.1, Fateev I.V.1, Muravyova T.I.1, Artemova K.G.1, Konstantinova I.D.1, Kuranova I.P.2,3, Miroshnikov A.I.1
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Affiliations:
- Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry
- Shubnikov Institute of Crystallography
- National Research Center “Kurchatov Institute”
- Issue: Vol 42, No 5 (2016)
- Pages: 512-521
- Section: Article
- URL: https://journals.rcsi.science/1068-1620/article/view/228124
- DOI: https://doi.org/10.1134/S1068162016040075
- ID: 228124
Cite item
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.
About the authors
R. S. Esipov
Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry
Author for correspondence.
Email: esipov@ibch.ru
Russian Federation, ul. Miklukho-Maklaya 16/10, Moscow, 117997
Yu. A. Abramchik
Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry
Email: esipov@ibch.ru
Russian Federation, ul. Miklukho-Maklaya 16/10, Moscow, 117997
I. V. Fateev
Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry
Email: esipov@ibch.ru
Russian Federation, ul. Miklukho-Maklaya 16/10, Moscow, 117997
T. I. Muravyova
Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry
Email: esipov@ibch.ru
Russian Federation, ul. Miklukho-Maklaya 16/10, Moscow, 117997
K. G. Artemova
Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry
Email: esipov@ibch.ru
Russian Federation, ul. Miklukho-Maklaya 16/10, Moscow, 117997
I. D. Konstantinova
Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry
Email: esipov@ibch.ru
Russian Federation, ul. Miklukho-Maklaya 16/10, Moscow, 117997
I. P. Kuranova
Shubnikov Institute of Crystallography; National Research Center “Kurchatov Institute”
Email: esipov@ibch.ru
Russian Federation, Leninski pr. 59, Moscow, 119333; pl. akad. Kurchatova 1, Moscow, 123182
A. I. Miroshnikov
Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry
Email: esipov@ibch.ru
Russian Federation, ul. Miklukho-Maklaya 16/10, Moscow, 117997