Effect of Enzyme Concentration on Enzyme Activity

1. 1. 1. Add 5 ml of water to an empty tube (this is the BLANK)
      2. Add 5 ml of pH 7 buffer to 3 separate tubes
      3. Follow the dilution scheme below:

1. 1. 4. In 4 separate tubes, ADD 4 ml of starch solution
         • label them 1x, 1/5x, 1/25x, 1/125x
      5. Add 2 drops of iodine to each starch tube and the Blank
      6. Read the Blank in the spectrophotometer and calibrate it to 100% transmittance at 560nm
      7. Read each tube in the spectrophotometer. This is time 0 min
      8. Add 35 drops of diluted amylase solutions to the appropriately labeld tubes simultaneously and mix.
         • Each tube receives a different Amylase dilution
      9. At 2 minute intervals, quickly read ALL tubes in the spectrophotometer.
      10. Continue reading the samples every 2 minutes until you reach 22 minutes on the table below.

Time (min)	1X % Trans	1/5X % Trans	1/25X % Trans	1/125X % Trans
0
2
4
6
8
10
12
14
16
18
20
22

Tags: quantitative reasoning, analysis, inquiry, integration of knowledge