Name: Sawong Kusuma

Exercise 8: Chemical and Physical Processes of Digestion: Activity 3: Assessing Pepsin Digestion of Protein Lab Report

Pre-lab Quiz Results

You scored 100% by answering 5 out of 5 questions correctly.

1. Where in the body does protein digestion begin?

You correctly answered: b. the stomach

2. The substrate for pepsin is

You correctly answered: d. protein and peptides.

3. In this activity the substrate you will be using to detect protein digestion is
You correctly answered: d. BAPNA.

4. Negative results with the negative controls

You correctly answered: d. are expected and validate the experiment.

5. A spectrophometer measures
You correctly answered: c. optical density.

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Experiment Results
Predict Question:
Predict Question: At which pH do you think pepsin will have the highest activity?
Your answer : a. pH 2.0

Stop & Think Questions:

Which of the following tubes are negative control tubes?
You correctly answered: e. tubes 3 and 4

An optical density greater than zero indicates that

You correctly answered: e. BAPNA digestion has occurred and pepsin is active.

Experiment Data:

Tube No. Reagent 1 Reagent 2 Reagent 3 Boiled Time Temp. Optical

Density
1 Pepsin BAPNA pH 2.0 Buffer + 60 37 0.00
2 Pepsin BAPNA pH 2.0 Buffer - 60 37 0.40
3 Pepsin DeIonized Water pH 2.0 Buffer - 60 37 0.00
4 DeIonized Water BAPNA pH 2.0 Buffer - 60 37 0.00
5 Pepsin BAPNA pH 7.0 Buffer - 60 37 0.03
6 Pepsin BAPNA pH 9.0 Buffer - 60 37 0.00

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Post-lab Quiz Results
You scored 100% by answering 4 out of 4 questions correctly.

1. Pepsin would be most active

You correctly answered: b. in the stomach.

2. Which two tubes validated the results of the experiment?

You correctly answered: c. tubes 3 and 4

3. With more enzyme activity the optical density

You correctly answered: a. increased.

4. If pepsin were digesting an actual protein substrate, the product would be

You correctly answered: c. peptides and amino acids.

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Review Sheet Results
1. Describe the effect that boiling had on pepsin and how you could tell that it had that effect.
Your answer:
Pemanasan yang melebihi titik optimum suatu protein dapat membuat struktur tiga dimensinya berubah, dalam hal ini
enzim pepsin. Perubahan juga meliputi perubahan pada sisi aktif enzim, tempat berikatan dengan substrat sehingga
membuat enzim tidak dapat bekerja atau dengan istilah lain terdenaturasi. [1] Dalam eksperimen, ini ditunjukkan dari hasil
tabung 1 yang sebelumnya dipanaskan, dengan hasil tidak terjadi aktivitas pepsin dimana BAPNA tidak melepaskan
yellow dye product dilihat dari nilai optical density-nya yang nol (0). Sedangkan pada campuran reagen yang sama namun
tidak dipanaskan yakni tabung 2, menghasilkan optical density 0.40.

2. Was your prediction correct about the optimal pH for pepsin activity? Discuss the physiological correlation behind your
results.
Your answer:
Seusai dengan prediksi saya, hasil ini dapat disimpulkan dengan membandingkan optical density pada tabung 2, 5, dan 6,
yang memiliki variabel bebas berupa pH. Pada tabung 9, optical density 0, sedangkan pada tabung 5 bernilai 0.03 dan pada
tabung 2 bernilai 0.04. Hasil ini menunjukkan pH optimal untuk aktivitas pepsin adalah pH 2.0. Ini sesuai dengan fakta
fisiologis, dimana pepsin bekerja di lambung manusia dengan suasana sangat asam, dengan pH 1.8-3.5. [2]

3. What do you think would happen if you reduce the incubation time to 30 minutes for tube 5?
Your answer:
Jumlah produk yang dikatalis oleh enzim akan berbanding lurus dengan waktu atau lamanya reaksi berlangsung.
Dengan demikian, jika waktu inkubasi diturunkan dari 30 menit ke 5 menit pada tabung 5 menit, maka yang pasti akan
terjadi penurunan nilai optical density yang menunjukkan berkurangnya jumlah BAPNA yang dikatalis oleh pepsin.

Citation:
[1] Sherwood L. Fisiologi manusia: dari sel ke sistem, Ed. 8. Jakarta: Penerbit Buku Kedokteran EGC; 2012.
[2] Guyton, A. C., Hall, J. E., Text book of medical physiology, 12th Ed. Philadelpia, PA: Saunders Elsevier; 2011.

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