Abstract
The activity of NADP+-specific isocitrate dehydrogenase (NADP+-IDH, EC 1.1.1.42) was investigated during the ripening of tomato (Lycopersicon esculentum Mill.) fruit. In the breaker stage, NADP+-IDH activity declined but a substantial recovery was observed in the late ripening stages when most lycopene synthesis occurs. These changes resulted in higher NADP+-IDH activity and specific polypeptide abundance in ripe than in green fruit pericarp. Most of the enzyme corresponded to the predominant cytosolic isoform which was purified from both green and ripe fruits. Fruit NADP+-IDH seems to be a dimeric enzyme having a subunit size of 48 kDa. The K m values of the enzymes from green and ripe pericarp for NADP+, isocitrate and Mg2+ were not significantly different. The similar molecular and kinetic properties and chromatographic behaviour of the enzymes from the two kinds of tissue strongly suggest that the ripening process is not accompanied by a change in isoenzyme complement. The increase in NADP+-IDH in the late stage of ripening also suggests that this enzyme is involved in the metabolism of C6 organic acids and in glutamate accumulation in ripe tissues.
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Abbreviations
- DEAE:
-
diethylaminoethyl
- GADPH:
-
NADP+-dependent glyceraldehyde 3-phosphate dehydrogenase
- GOGAT:
-
glutamate synthase
- GS:
-
glutamine synthetase
- Mr :
-
relative molecular mass
- NAD+-IDH:
-
NAD+-specific isocitrate dehydrogenase
- NADP+ -IDH:
-
NADP+-specific isocitrate dehydrogenase
- PEPCase:
-
phosphoenolpyruvate carboxylase
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We thank to M. Hodges (Université Paris Sud) for the revision and criticism of the manuscript. This work was partially supported by a Grant from Direction General de Investigatión Científica y Técnica (PB 92-0423) to F.M.C. F.G. was supported by a fellowship from the Programa General de Formatión de Personal Investigador, Junta de Andalucia, and by a short-term fellowship from Ministerio de Educacion y Ciencia. S.G. is recipient of a fellowship from the Programa de Formatión de Personal Investigador en el Extranjero; Ministerio de Educación y Ciencia, Spain.
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Gallardo, F., Gálvez, S., Gadal, P. et al. Changes in NADP+-linked isocitrate dehydrogenase during tomato fruit ripening. Planta 196, 148–154 (1995). https://doi.org/10.1007/BF00193228
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DOI: https://doi.org/10.1007/BF00193228