Purification and characterisation of a soluble nucleotide pyrophosphatase/phosphodiesterase from prickly pear (Opuntia ficus indica) fruits

A glycosylated metallo-protein, nucleotide pyrophosphatase/phosphodiesterase, in a soluble form was purified to homogeneity from prickly pear (Opuntia ficus indica) fruits. The native protein had a molecular mass of 105 ± 8 kDa and was formed by two apparently identical subunits each containing 1 Ca2+ and 1 Mg2+ ion. The Opuntia enzyme exhibited hydrolytic activities toward pyrophosphate/phosphodiester bonds of a broad range of natural substrates, but among these, only NAD(P) and NAD(P)H were hydrolysed very efficiently. Moreover, Opuntia pyrophosphatase/phosphodiesterase hydrolysed the artificial substrate thymidine 5′-monophosphate 4-nitrophenyl ester, whereas it did not show any catalytic activity toward bis-4-nitrophenyl phosphate, which is a substrate of other pyrophosphatase/phosphodiesterase enzymes. We observed an increase of enzyme activity from the green to the red stage of fruits development, suggesting that ONPP activity might be related to the ripening of prickly pears. The protein was shown to be resistant to 75 °C for 30 min. Other biochemical characteristics were investigated and are reported here.