Partial purification and some regulatory properties of pyruvate kinase from germinating castor bean endosperm

Abstract
The activity of pyruvate kinase in castor bean endosperm was found to increase rapidly from the 3rd to 4th day of germination. The crude enzyme is very labile, but, was stabilized by adding Mg2+ with a high concentration of ethyleneglycol or glycerol. The enzyme thus stabilized was partially purified about 150-fold by fractionation with ammonium sulfate, and chromatography with DEAE-cellulose and Sephadex G-150. The enzyme showed normal Michaelis-Menten kinetics for both substrates; Km for PEP and ADP were 0.058 mM and 0.13 mM, respectively. Optimum pH was near 6.5. The effects of many kinds of amino acids, intermediates of glycolysis, and the tricarboxylic acid cycle on the activity of the enzyme were tested. ATP was found to inhibit and α-ketoglutarate to activate enzymatic activity.

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