De Novo Synthesis of Isocitritase in Peanut (Arachis hypogaea L.) Cotyledons

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Abstract
Germination of peanut seeld is accompained by a rapid increase in isocitritase (isocitrate lyase, EC 4.1.3.1) during the first 4 days. The presence of cycloheximide (50 [mu]g/ml) during water imbibition inhibited the increase in Isocitriase activity. Actinomycin D conversely did not inhibit isocitritase activity until the second day of imbibition while RNA synthesis was inhibited. Germination of peanut seed in 14C-reconstituted amino acids followed by fractionation of a 20 to 35% ammonium sulfate preparation on a Sephadex G-200 column (57-fold purification) showed that the active enzymic fraction coincided with a large peak of radioactivity. Germination of peanut seed in 45% D2O followed by enzyme purification and CsCl equilibrium centrifugation revealed that all the enzyme from D2O seed had a higher density than normal isocitritase. These data indicate that isocitritase in peanut seed is synthesized de novo.

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