PLEASE SPECIFY THE FUNCTIONS AND ENZYMES PRESENT FOR PROTEIN DIGESTION
Biochemistry
9th Edition
ISBN:9781319114671
Author:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.
Publisher:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.
Chapter1: Biochemistry: An Evolving Science
Section: Chapter Questions
Problem 1P
Related questions
Question
PLEASE SPECIFY THE FUNCTIONS AND ENZYMES PRESENT FOR PROTEIN DIGESTION
Procedure:
- Peptic digestion
- Prepare 4 test tubes as follows:
Tube 1 - 3 ml pepsin
Tube 2 - 3 ml of 0.4% HCl
Tube 3 - 3 ml pepsin + 1.5 ml of 0.4% HCl
Tube 4 - 3 ml pepsin + 1.5 ml of 1% Na2CO3
- Add equal slices of hard-boiled egg white to each tube.
- Cover and incubate at 40oC for 2 hours.
- Add 4 drops of toluene and store till the next lab period.
- Determine the extent of digestion by visually noting the size of the hard-boiled egg white in each tube.
- Withdraw 1 ml of the supernatant liquid from each tube and place in 4 separate test tubes labelled tube 1a. 2a, 3a, 4a.
- Neutralize tubes 2a& 3a with solid sodium carbonate.
- To each supernatant liquid, add 1 ml of 10% NaOH and 2 drops of 1% CuSO4. Compare the colors obtained in the 4 tubes.
- Place 1 ml of 0.5% peptone in a test tube and add 1 ml of 10% NaOH and 2 drops of 1% CuSO4. Compare the result of these tubes with the other 4 tubes above.
- Pancreatin digestion
- Prepare 4 test tubes as follows:
Tube 1 - 3 ml neutral pancreatin
Tube 2 - 3 ml water
Tube 3 - 3 ml water + 1.5 ml pancreatin
Tube 4 - 3 ml water + 1.5 ml pancreatin + 1.5 ml 1% Na2CO3
- Place equal size of hard-boiled egg white in all tubes.
- Place all tubes in water bath with a temperature maintained at 40oC for 1.5 hours.
- Add 3 drops of toluene in all tubes and incubate till the next lab period.
- Observe the size of the egg white in each of the tubes.
- Take 1 ml of the supernatant in each tube and place in 4 separate test tubes labelled 1b,2b,3b,4b. Add 1 ml of 10% NaOH and 2 drops of 1% CuSO4 in each tube. Mix and compare the result of the 4 tubes.
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