Use of potato tuber nucleotide pyrophosphatase to synthesize adenosine 5′-monophosphate methyl ester: Evidence that the solvolytic preferences of the enzyme are regulated by pH and temperature
✍ Scribed by Antonio Agudo; João Meireles Ribeiro; José Canales; José Carlos Cameselle
- Publisher
- John Wiley and Sons
- Year
- 1998
- Tongue
- English
- Weight
- 113 KB
- Volume
- 59
- Category
- Article
- ISSN
- 0006-3592
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✦ Synopsis
Nucleotide alkyl esters are pharmacologically important as potential (ant)agonists of purinoceptors and inhibitors of enzymes. Potato nucleotide pyrophosphatase (PNP) was compared with snake venom phosphodiesterase (SVP) as a catalyst to synthesize nucleotide alkyl esters. In methanol-water mixtures, the methanolysis/hydrolysis ratio of PNP, but not SVP, changed with pH and temperature, being optimal at high pH and low temperature. In a semi-preparative experiment, a crude PNP preparation produced 0.17 mM AMP-Omethyl ester (AMP-OMe) from 1 mM diadenosine 5Ј,5ٞ-P 1 ,P 2 -diphosphate (AppA) and 5M methanol, at pH 9 and 0°C. Drawbacks to large-scale use are: low rates inherent to low temperatures, ATP unsuitability as a substrate for alcoholysis, and high cost of AppA. Advantages of PNP vs. SVP are cheapness, non-toxicity, and availability of the enzyme source.