A simple, rapid extraction and assay procedure for the NAD+-dependent sorbitol dehydrogenase (SDH) in peach

Sorbitol is the major photosynthetic product in peach. In sink tissues, sorbitol is converted to fructose via the NAD+-dependent sorbitol dehydrogenase (SDH). A new procedure is described that allows rapid, simple quantification of SDH activity in growing tissues. The procedure uses only 0.01 to 2.5 grams of fresh tissue per sample, such that a single shoot tip, a single root tip, or about 5 g of fruit flesh can be assayed for SDH activity. Storage of samples at 4 or -20 C° over night resulted in significant loss of enzyme activity. Thus, freshly harvested tissues were ground with sand in buffer at 2°C in a mortar and pestle, and the homogenate centrifuged @ 3000 ´ g to remove particulate m…