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Cloning of Nucleoside Phosphorylase Genes from the Extremophilic Bacterium Halomonas chromatireducens AGD 8-3 with the Construction of Recombinant Producer Strains of These Proteins and the Study of Their Enzymatic Properties

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Abstract

Thymidine phosphorylase (deoA) and purine nucleoside phosphorylase (deoD) genes from the extremophilic bacterium Halomonas chromatireducens AGD 8-3 have been cloned. Expression plasmids were constructed, and highly efficient recombinant producer strains were obtained for these proteins. Recombinant nucleoside phosphorylases were isolated by ion-exchange chromatography in a homogeneous state, and their physical and enzymatic properties were studied. It was shown that the studied thymidine phosphorylase (HrTPP) and purine nucleoside phosphorylase (HrPNP) form the dimeric and hexameric forms, respectively. It was shown that the specific activity of HrTPP from the extremophilic bacterium H. chromatireducens AGD 8-3 was higher relative to thymidine (in comparison with its counterpart from E. coli).

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ACKNOWLEDGMENTS

The equipment of the Industrial Biotechnologies Center for Collective Use of the Fundamentals of Biotechnology Federal Research Center of the Russian Academy of Sciences was used in the study.

Funding

The study was partially supported by the Russian Foundation for Basic Research (project no. 18-04-00784 A).

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Authors and Affiliations

  1. Bach Institute of Biochemistry, Federal Research Center of Biotechnology, Russian Academy of Sciences, 119071, Moscow, Russia

    A. N. Antipov, N. N. Mordkovich, N. A. Okorokova & V. P. Veiko

  2. Winogradsky Institute of Microbiology, Federal Research Center of Biotechnology, Russian Academy of Sciences, 119071, Moscow, Russia

    T. V. Khijniak

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  1. A. N. Antipov
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Correspondence to V. P. Veiko.

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The authors declare that they have no conflict of interest. This article does not contain any studies involving animals or human participants performed by any of the authors.

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Translated by E. Makeeva

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Antipov, A.N., Mordkovich, N.N., Khijniak, T.V. et al. Cloning of Nucleoside Phosphorylase Genes from the Extremophilic Bacterium Halomonas chromatireducens AGD 8-3 with the Construction of Recombinant Producer Strains of These Proteins and the Study of Their Enzymatic Properties. Appl Biochem Microbiol 56, 37–43 (2020). https://doi.org/10.1134/S0003683820010020

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