BCH4160/2011:Notebook/Laura Lee/2011/11/08

Enzyme Kinetics

The goal of this experiment was to measure the kinetics of the digestion of starch by cellobiase with and without urea and betaine monohydrate in solution.

Materials

In this experiment we used:

• 1.2% stock starch solution
• 9 molal stock urea solution
• 10X stock resuspension buffer (pH = 5)
• Distilled water
• Iodine
• Cellobiase
• UV spectrophotometer

Procedure

This is an adapted protocol from previous students' studies. Refer to Tiffany Byerly's previous protocol and data.

Solution Preparation

1. Prepare 1X resuspension buffer from original 10X solution.
   • Dissolve 16.7mL original solution in 150mL distilled water.
2. Prepare 1.2% starch solution.
   • Dissolve 1.2g starch in 100mL buffer.
3. Prepare 9 molal (mol/kg) urea.
   • Dissolve 2.7g urea in 5mL buffer.
4. Prepare 4 molal betaine monohydrate.
   • Dissolve 2.7g urea in 5mL buffer.

This protocol is continued in Enzyme Kinetics part 2.

Data/Results

Refer to Enzyme Kinetics part 2.

Contact

This experiment was done with High Point University students Jigesh Patel and Brittany Webster.