In the first experiment, optimal temperature for enzymatic activity was tested. Five clean spectrophotometer tubes wereare necessary with the different temperatures labeled on them using a wax pencil (Blank, Room Temp, 35 degrees Celsius, 45 degrees Celsius, and 55 degrees Celsius). Then 1mL of …show more content…
Again, five clean spectrophotometer tubes were necessary with the different pH numbers labeled on them using a wax pencil (3, 4, 5, 6, and 8), and 1 mL of the buffers with those pH levels was added. Each tube needed 1 mL of substrate mixture which was added. Then the spectrophotometer to 470nm and the first spectrophotometer tube was wiped to be used to blank the spectrophotometer. Then 0.1 mL of enzyme from vial E1 ( 1x10^-7 M peroxidase) was added, then it was shaken with parafilm covering the top. The tube was quickly placed into the spectrophotometer, and it was necessary to be sure not to spill any of the reaction mixture or this experiment would have to be repeated. The absorbance was recorded every 5 seconds for 60 seconds and this process was repeated for each spectrophotometer tube that went in one at a