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Characterization of a recombinant cold-adapted purine nucleoside phosphorylase and its application in ribavirin bioconversion

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Abstract

Ribavirin is a broad-spectrum antiviral drug and can be produced by enzymatic synthesis by purine nucleoside phosphorylase (PNP). In this study, we describe the application of such a cold-adapted XmPNP in ribavirin bioconversion which showed approximately 15°C lower optimum temperature and 1.80-fold higher catalytic efficiency (kcat/Km) at 37°C within substrate inosine than homolog in E. coli. By contrast, E. coli (XmPNP) took only 12 h to reach maximum substrate conversion rate (70%) under its optimum temperature (50°C) by using recombinant strain cell as enzyme source, but E. coli (EcPNP) did at 24 h. These results suggest cold-adapted PNP is one attractive candidate for ribavirin bioconversion and other nucleoside medications to improve the catalytic efficiency.

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Abbreviations

XmPNP:

Pseudoalteromonas sp. XM2107 purine nucleoside phosphorylas

EcPNP:

E. coli purine nucleoside phosphorylas

E. coli (EcPNP):

E. coli BL21 (DE3) with recombinant vector pHEC

E. coli (XmPNP):

E. coli BL21 (DE3) with recombinant vector pHXM

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Acknowledgments

We sincerely thank Dr. Zeng Ruiying of the Polar Reasearch Institute of China for the donation of Pseudoalteromonas sp. This work was supported by the High Technology Research and Development Program of Tianjin High School (Grant No. 20070903).

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Authors and Affiliations

  1. College of Biotechnology, Tianjin University of Science and Technology, Key Laboratory of Industrial Microbiology of Education Ministry, Tianjin, 300457, China

    Xixian Xie, Guanglu Wang & Jungang Xia

  2. Metabolic Engineering Laboratory, College of Biotechnology, Tianjin University of Science and Technology, No. 29, 13 Main Street, Tianjin Economic and Technological Development Zone, Tianjin, 300457, China

    Ning Chen

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  1. Xixian Xie
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  2. Guanglu Wang
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  3. Jungang Xia
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  4. Ning Chen
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Correspondence to Ning Chen.

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Xixian Xie and Guanglu Wang contributed equally to this work.

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Xie, X., Wang, G., Xia, J. et al. Characterization of a recombinant cold-adapted purine nucleoside phosphorylase and its application in ribavirin bioconversion. World J Microbiol Biotechnol 27, 1175–1181 (2011). https://doi.org/10.1007/s11274-010-0564-7

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  • DOI: https://doi.org/10.1007/s11274-010-0564-7

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