Biotechnology Letters

, Volume 39, Issue 12, pp 1903–1910 | Cite as

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

Original Research Paper



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


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%.


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


Gene mining Halogenated nucleoside Pyrimidine nucleoside phosphorylase 



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.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

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

Supplementary material

10529_2017_2423_MOESM1_ESM.doc (492 kb)
Supplementary material 1 (DOC 492 kb).


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Copyright information

© Springer Science+Business Media B.V. 2017

Authors and Affiliations

  1. 1.College of Biotechnology and Pharmaceutical EngineeringNanjing Tech UniversityNanjingChina
  2. 2.School of Pharmaceutical SciencesNanjing Tech UniversityNanjingChina
  3. 3.Department of Hepatobiliary and Pancreatic Surgery, Zhongda Hospital, Medical SchoolSoutheast UniversityNanjingChina

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