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A thermostable pyrimidine nucleoside phosphorylase from Brevibacillus borstelensis LK01 for synthesizing halogenated nucleosides

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Abstract

Objective

To isolate a thermostable pyrimidine nucleoside phosphorylase (PyNP) from mesophilic bacteria by gene mining.

Results

BbPyNP from Brevibacillus borstelensis LK01 was isolated by gene mining. BbPyNP had a highest 60% identity with that of reported PyNPs. BbPyNP could catalyze the phosphorolysis of thymidine, 2′-deoxyuridine, uridine and 5-methyuridine. BbPyNP had good thermostability and retained 73% of its original activity after 2 h incubation at 50 °C. BbPyNP had the highest activity at an optimum alkaline pH of 8.5. BbPyNP was stable from pH 7 to 9.8. Under preliminary optimized conditions, the biosynthesis of various 5-halogenated pyrimidine nucleosides by BbPyNP reached the yield of 61–84%.

Conclusion

An efficient approach was estimated in isolating thermostable PyNP from mesophilic bacteria.

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (81673321, 21376119, 21506099), and Natural Science Foundation of the Jiangsu Higher Education Institution of China (15KJB530008).

Supporting information

Supplementary Table 1—The primers used in the experiments.

Supplementary Fig. 1—Mass spectrometry analysis of 5-fluoro-2′-deoxyuridine.

Supplementary Fig. 2—1H-NMR spectroscopic data of 5-fluoro-2′-deoxyuridine.

Supplementary Fig. 3—13C-NMR spectroscopic data of 5-fluoro-2′-deoxyuridine.

Author information

Authors and Affiliations

  1. College of Biotechnology and Pharmaceutical Engineering, Nan**g Tech University, Nan**g, China

    Ke Liu, Youzhi Zhou, **song Zhang & Bingfang He

  2. School of Pharmaceutical Sciences, Nan**g Tech University, Nan**g, China

    Jianlin Chu & Bingfang He

  3. Department of Hepatobiliary and Pancreatic Surgery, Zhongda Hospital, Medical School, Southeast University, Nan**g, China

    Yewei Zhang

Authors
  1. Ke Liu
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  2. Youzhi Zhou
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  3. **song Zhang
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  4. Jianlin Chu
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  5. Yewei Zhang
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  6. Bingfang He
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Corresponding authors

Correspondence to Yewei Zhang or Bingfang He.

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The authors declare that they have no conflict of interest.

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This article does not contain any studies with human participants or animals performed by any of the author.

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Supplementary material 1 (DOC 492 kb).

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Liu, K., Zhou, Y., Zhang, J. et al. A thermostable pyrimidine nucleoside phosphorylase from Brevibacillus borstelensis LK01 for synthesizing halogenated nucleosides. Biotechnol Lett 39, 1903–1910 (2017). https://doi.org/10.1007/s10529-017-2423-1

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  • DOI: https://doi.org/10.1007/s10529-017-2423-1

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