Assay of PDHc from Plant

The powder of mitochondria in acetone was resuspended in 25 mM TES buffer (pH 7.4) containing 200 pM TPP (thiamine pyrophosphate), 5 mM DTT (dithio-threitol), and 2 mM MgS04 at a final concentration of 20 mg/mL, and undissolved material was removed at 27,000 g for 15 min. 

Six samples of enzyme solution (0.5 mL) were taken. One of them was added to water as a control, and the rest were mixed with test compounds at a final concentration 10 , 10 10, 10 and 10 g/mL respectively. The mixture was 

1 Assay of PDHc from Mung Bean (in Vitro) 

The inhibition of enzyme from mung bean was tested according to the same method as that used for pea. 

000 g for 45 min. The surface of the pellet was washed with ice water, then resuspended in acetone at -20 °C, centrifuged at 27,000 g for 20 min and resuspended and centrifuged for additional three times in cold acetone. The final pellet was dried with a stream of air and stored at -20 °C. 

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