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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
Author(s) -
Agudo Antonio,
Ribeiro João Meireles,
Canales José,
Cameselle José Carlos
Publication year - 1998
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/(sici)1097-0290(19980705)59:1<62::aid-bit8>3.0.co;2-r
Subject(s) - pyrophosphatase , chemistry , nucleotide , enzyme , adenosine monophosphate , inorganic pyrophosphatase , adenosine , biochemistry , pyrophosphate , gene
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 m M AMP‐O‐methyl ester (AMP‐OMe) from 1 m M diadenosine 5′,5‴‐ P 1 , P 2 ‐diphosphate (AppA) and 5 M 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. © 1998 John Wiley & Sons, Inc. Biotechnol Bioeng 59:62–67, 1998.