place brown iodine into wells in a spotting tile, put equal volume of starch (substrate) into 5 test tubes, add a set volume of a pH buffer to each tube, ensure each tube has a different pH, place in a water bath at body temperature for 5 minutes, place a set volume of the enzyme amylase into each test tube, stir and keep in the water bath, remove a sample of the mixture from each tube and place a drop onto the iodine, do this every 30 seconds, record the time it takes for the starch to STOP turning blue black for each pH, This indicates that all the starch has been broken down by the amylase.

enzymes required practical

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