Plant nucleotide pyrophosphatase/phosphodiesterase (NPPase), method of
production, use in the manufacture of testing devices and in the
production of transgenic plants. NPPase is an enzyme that catalyses the
hydrolysis of a wide range of small molecules with phosphodiester and
phosphosulphate bonds, in particular ADPG (adenosine diphosphate glucose)
and APS (adenosine 5'-phosphosulphate). The enzyme obtained from plant
extracts is used in assay devices for determining levels of nucleoside
diphosphate sugars, based either on the sugar-1-phosphate released, or on
the nucleoside monophosphate, both of which are products formed by the
reaction catalysed by NPPase, as well as the detection of
sulphonucleotides such as 3'-phosphoadenosine 5'-phosphosulphate (PAPS)
and APS. The amino acid sequence of the enzyme is also described, as well
as the nucleotide sequence of a complete cDNA and another incomplete
cDNA. Finally, it describes the production of transgenic plants that
overexpress NPPase and that have a high content of sugars, low content of
starch and cell-wall polysaccharides and high resistance to high
concentrations of salts and high temperature.