Molecules 2013, 18(10), 12587-12598; doi:10.3390/molecules181012587
Article

Enzymatic Synthesis of Highly Fluorescent 8-Azapurine Ribosides Using a Purine Nucleoside Phosphorylase Reverse Reaction: Variable Ribosylation Sites

1 Department of Biophysics, University of Varmia & Masuria, 4 Oczapowskiego St., 10-719 Olsztyn, Poland 2 Division of Biophysics, Institute of Experimental Physics, University of Warsaw, Zwirki i Wigury 93, 02-089 Warsaw, Poland 3 Division of Physical Chemistry, Rudjer Bošković Institute, POB 180, HR-10002 Zagreb, Croatia
* Author to whom correspondence should be addressed.
Received: 30 August 2013; in revised form: 25 September 2013 / Accepted: 30 September 2013 / Published: 11 October 2013
(This article belongs to the Special Issue Synthesis of Nucleosides, Nucleotides and Their Derivatives)
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Abstract: Various forms of purine-nucleoside phosphorylase (PNP) were used as catalysts of enzymatic ribosylation of selected fluorescent 8-azapurines. It was found that the recombinant calf PNP catalyzes ribosylation of 2,6-diamino-8-azapurine in a phosphate-free medium, with ribose-1-phosphate as ribose donor, but the ribosylation site is predominantly N7 and N8, with the proportion of N8/N7 ribosylated products markedly dependent on the reaction conditions. Both products are fluorescent. Application of the E. coli PNP gave a mixture of N8 and N9-substituted ribosides. Fluorescence of the ribosylated 2,6-diamino-8-azapurine has been briefly characterized. The highest quantum yield, ~0.9, was obtained for N9-β-d-riboside (λmax 365 nm), while for N8-β-d-riboside, emitting at ~430 nm, the fluorescence quantum yield was found to be close to 0.4. Ribosylation of 8-azaguanine with calf PNP as a catalyst goes exclusively to N9. By contrast, the E. coli PNP ribosylates 8-azaGua predominantly at N9, with minor, but highly fluorescent products ribosylated at N8/N7.
Keywords: 8-azapurines; nucleosides; enzymatic synthesis; fluorescence; purine-nucleoside phosphorylase

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MDPI and ACS Style

Stachelska-Wierzchowska, A.; Wierzchowski, J.; Wielgus-Kutrowska, B.; Mikleušević, G. Enzymatic Synthesis of Highly Fluorescent 8-Azapurine Ribosides Using a Purine Nucleoside Phosphorylase Reverse Reaction: Variable Ribosylation Sites. Molecules 2013, 18, 12587-12598.

AMA Style

Stachelska-Wierzchowska A, Wierzchowski J, Wielgus-Kutrowska B, Mikleušević G. Enzymatic Synthesis of Highly Fluorescent 8-Azapurine Ribosides Using a Purine Nucleoside Phosphorylase Reverse Reaction: Variable Ribosylation Sites. Molecules. 2013; 18(10):12587-12598.

Chicago/Turabian Style

Stachelska-Wierzchowska, Alicja; Wierzchowski, Jacek; Wielgus-Kutrowska, Beata; Mikleušević, Goran. 2013. "Enzymatic Synthesis of Highly Fluorescent 8-Azapurine Ribosides Using a Purine Nucleoside Phosphorylase Reverse Reaction: Variable Ribosylation Sites." Molecules 18, no. 10: 12587-12598.

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