Effect of Temperature on Enzyme Activity
- Add 5 ml of H2O to a tube (this is the BLANK)
- Add 5 ml of starch (substrate) to 3 separate tubes
- one on ice (0°C), one on the bench (25°C) and one in a 40°C water bath
- Add 2 drops of iodine to each tube and mix: Blank, 0°C, 25°C, 40°C
- Read the Blank in the spectrophotometer and calibrate it to 100% transmittance at 560nm
- Read each tube in the spectrophotometer. This is time 0 min.
- Add 35 drops of amylase solution to each tube simultaneously, mix and ensure incubation is occurring at the correct temperature.
- At 2 minute intervals, quickly read ALL tubes in the spectrophotometer and immediately return the tubes to the proper temperature.
- Continue reading the samples every 2 minutes until you reach 22 minutes on the table below.
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