ENZYME ROR

• Protease enzyme


  Tripin, produced by the small intestine of most mammals, involved in the hydrolysis of peptide bonds in proteins
• Substrate


  Casine, the protein in milk
• Enzyme-controlled reaction


  Casine digested by Tripin to produce smaller, more soluble peptides
• Preparing solutions


  • Use a water bath to heat solutions to 40°C
  • Prepare 10 mL of 3% powdered milk solution in two test tubes, one labeled T and one labeled C
  • Prepare 4 mL of pH 7 buffer in the C tube, and 2 mL of pH 7 buffer plus 2 mL of 0.5% Tripin solution in the T tube
• Procedure


  1. Place the test tubes in the water bath for 5 minutes to reach 40°C

  2. Add the Trypsin solution to the T tube and mix

  3. Add the milk solution to the C tube and mix

  4. Start the timer immediately after mixing

  5. Observe the tubes and stop the timer when the X mark becomes visible through the T tube
• Endpoint


  The point at which the X mark becomes visible through the T tube
• Calculating reaction rate


  Divide the total reaction time by the time taken for the reaction to complete
• Increasing temperature


  Increases the reaction rate up to an optimum temperature
• Temperatures above optimum


  Decrease the reaction rate
• The exact optimum temperature cannot be determined with certainty from the data