title: 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.
abstract: 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-O-methyl ester (AMP-OMe) from 1 mM diadenosine 5',5"'-P1,P2-diphosphate (AppA) and 5M methanol, at pH 9 and 0 degrees 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.
authors: Unidad de Bioquímica y Biología Molecular, Facultad de Medicina, Universidad de Extremadura, Apartado de Correos 108, E-06080 Badajoz, Spain.
authors: A
authors: A
authors: Agudo
authors: J M
authors: JM
authors: Ribeiro
authors: J
authors: J
authors: Canales
authors: J C
authors: JC
authors: Cameselle
mesh_terms: D000249:Adenosine Monophosphate / Q000031:analogs & derivatives / Q000138:chemical synthesis*; D002851:Chromatography, High Pressure Liquid; D006863:Hydrogen-Ion Concentration; D007202:Indicators and Reagents; D007700:Kinetics; D043264:Phosphodiesterase I; D010727:Phosphoric Diester Hydrolases / Q000302:isolation & purification / Q000378:metabolism; D011755:Pyrophosphatases / Q000302:isolation & purification / Q000378:metabolism; D011198:Solanum tuberosum / Q000201:enzymology*; D012995:Solubility; D013816:Thermodynamics
chemical_list: D007202:Indicators and Reagents; D000249:Adenosine Monophosphate; D010727:Phosphoric Diester Hydrolases; D043264:Phosphodiesterase I; C470790:phosphodiesterase I, snake venom; D011755:Pyrophosphatases; C021899:nucleotide pyrophosphatase