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Issue 44, 2017
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Development of an enzyme cycling method by a purine nucleoside phosphorylase for assaying inorganic phosphate

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Abstract

We have developed a novel enzymatic cycling method that uses purine nucleoside phosphorylase (PNP) (EC 2.4.2.1) from Bacillus sp. to measure inorganic phosphate. The method utilizes the reversibility of the PNP reaction, in which the forward and reverse reactions are catalyzed in the presence of an excess amount of inosine and guanine, respectively, a principle similar to that previously demonstrated with creatine kinase (CK). Real-time detection was accomplished by coupling the reaction with commercially available xanthine dehydrogenase (EC 1.17.1.4) in the presence of NAD+. The efficiency of the cycling reaction per unit of enzyme (U mlāˆ’1) was remarkably higher than the CK.

Graphical abstract: Development of an enzyme cycling method by a purine nucleoside phosphorylase for assaying inorganic phosphate

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Publication details

The article was received on 22 Aug 2017, accepted on 27 Sep 2017 and first published on 03 Oct 2017


Article type: Paper
DOI: 10.1039/C7AY02016C
Citation: Anal. Methods, 2017,9, 6235-6239
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    Development of an enzyme cycling method by a purine nucleoside phosphorylase for assaying inorganic phosphate

    S. Ueda and S. Sakasegawa, Anal. Methods, 2017, 9, 6235
    DOI: 10.1039/C7AY02016C

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