A thermostable pyrimidine nucleoside phosphorylase from Brevibacillus borstelensis LK01 for synthesizing halogenated nucleosides
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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.
KeywordsGene 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).
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.
This article does not contain any studies with human participants or animals performed by any of the author.
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