BIOL-101.

602 GENERAL BIOLOGY I

MATH AND SCIENCES DEPARTMENT
Fall 2023

Lab Report

Name: Hamza Section: 602 Due Date:

You have one week to submit the lab report from the date of the experiment.

Please note that this lab activity is worth 20 points total where the breakdown is as follows:

Good lab behavior - abiding by the safety measures as instructed in the lab 2 Points

Question Answer Section 7 Points

Observation tables 8 Points

Conclusion 3 Points

Section 1: Answer the below questions.

1. Define four levels of protein organization. (4 Points)

Levels of proteins
1. Primary Structure: This is the specific sequence of amino acids in a protein, which is essential
for its unique identity and interactions with other molecules.

2. Secondary Structure: It involves local folding patterns in a protein chain, like alpha helices and
beta sheets, stabilized by hydrogen bonds.

3. Tertiary Structure: Tertiary structure is the three-dimensional arrangement of the entire protein,
influenced by various interactions like disulfide bonds, hydrogen bonds, and hydrophobic
interactions, crucial for its overall shape and function.

4. Quaternary Structure: Quaternary structure relates to the association of multiple protein chains
(subunits) to form a functional protein complex, determining its function in a biological
context.

5. What are the factors that affect enzyme activity? (3 Points)

Factors affecting enzyme activity

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 BIOL-101.602 GENERAL BIOLOGY I
MATH AND SCIENCES DEPARTMENT
Fall 2023

Lab Report

1. Temperature: Enzymes function best between a certain range of temperatures. Although

activity is increased by higher temperatures, severe heat can denature enzymes.

2. pH Level: There is a specific pH where enzymes work at their best. They can have
their activity disrupted by pH variations.

3. Substrate Concentration: The quantity of molecules in the substrate affects the activity of
the enzyme. Up until all active sites are saturated, greater substrate concentration initially
promotes activity.

Section 2: Fill the following observation tables.

Data Table 1: Enzyme Specificity

Well Plate Contents Glucose Concentration

(mmol/L)
(1 Point Each)

A 3 drops 20% glucose + 3 drops dH2O 60-100mmol/L

B 3 drops milk + 3 drops 0 mmol/L

C 3 drops dH2O + 3 drops lactase 0 mmol/l

D 3 drops milk + 3 drops lactase 5-15 mmol/L

Data Table 2: Enzyme and pH

Well Plate Contents Glucose Concentration

(mmol/L)
(1 Point Each)
5-15 mmol/L
1 3 drops pH 3.5 buffer + 2 drops lactase + 2 drops milk

2 3 drops pH 5.0 buffer + 2 drops lactase + 2 drops milk 5-15 mmol/L

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 BIOL-101.602 GENERAL BIOLOGY I
MATH AND SCIENCES DEPARTMENT
Fall 2023

Lab Report

0 mmol/L
3 3 drops pH 6.8 buffer + 2 drops lactase + 2 drops milk

0 mmol/L
4 3 drops pH 11.5 buffer + 2 drops lactase + 2 drops milk

Section 3: Conclusion

In the table below, explain what you can conclude from the experiments you performed.

Enzyme Specificity (1.5 Points) Enzyme and pH (1.5 Points)

Did enzyme react with all the solutions? What is the optimum pH for laccase activity?
Give reason  It seems that lactase functions most
 The enzyme did, in fact, partially react effectively in the pH range of 3.5 to
with each solution. Where lactase was 5.0. Lactase is efficient at converting
absent, but glucose was present, a lactose into glucose and galactose
reaction occurred, resulting in a glucose
within this pH range, as seen by the
concentration of 60–100 mmol/L in
Plate A. Although lactose is the enzyme
same glucose concentration range
lactase's main substrate, evidence (5-15 mmol/L) on Plates 1 (pH 3.5)
suggests that it also has modest activity and 2 (pH 5.0), both of which
on glucose. There was no reaction in contained lactase and milk. There
Plate B, which indicates that lactase was no response seen in Plate 3 (pH
only digests the lactose present in milk. 6.8), indicating that lactase activity
There was similarly no reaction in Plate is greatly hindered or decreased
C when dH2O was substituted for the under slightly alkaline
milk, proving that lactase needs lactose circumstances. No response was
to function. However, a reaction seen in Plate 4 (pH 11.5), proving
occurred in Plate D, where lactase and
that lactase is entirely inert and
milk were both present, resulting in a
glucose content of 5–15 mmol/L. The
denatured under excessively alkaline
number of molecules in the substrate circumstances. In conclusion, the
has an impact on the enzyme's activity. data suggest that the ideal pH range
Greater substrate concentration at first for lactase activity is between 3.5
stimulates activity until all active sites and 5.0.
are saturated.

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 BIOL-101.602 GENERAL BIOLOGY I
MATH AND SCIENCES DEPARTMENT
Fall 2023

Lab Report

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 BIOL-101.602 GENERAL BIOLOGY I
MATH AND SCIENCES DEPARTMENT
Fall 2023

Lab Report

Instructions

A. Enzymes and Specificity

1. Prepare the lactase solution as follows:

a) Use the graduated cylinder to measure and transfer 150 mL of distilled water into the plastic
cup labeled “lactase”.

b) Crush the lactase tablet. See Figure below.

Note: If there is a bit of crushed lactase that does not dissolve into the water that is alright.

2. Place all 3 prepared solutions onto your work surface. See Figure below.

3. Label 4 wells of the 12-well plate “A, B, C, and D” Then place the labeled well plate on top of the
paper towel.

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 BIOL-101.602 GENERAL BIOLOGY I
MATH AND SCIENCES DEPARTMENT
Fall 2023

Lab Report

4. Gather 5 glucose test strips and label the strips “A, B, C, and D”. Place each glucose strip in front
of its corresponding well. See Figure below.

5. As described in Data Table 1, use the 20% glucose solution bottle and labeled pipets to add
solutions to the following wells:

a) In Well A: add 3 drops of 20% glucose and 3 drops of dH2O

b) In Well B: add 3 drops of milk and 3 drops of dH2O

c) In Well C: add 3 drops of dH2O and 3 drops of lactase

d) In Well D: add 3 drops of milk and 3 drops of lactase

Note: Ensure that pipets are only used to transfer solutions from their designated cups.

6. Allow each of the reactions to occur for 5 minutes.

7. After 5 minutes, pick up the glucose strip labeled “A”, and insert the green end into the reaction in
Well A for 5 seconds.

8. After 5 seconds, remove the strip from the well and place it on the paper towel.

9. Use the Glucose Color Chart shown below to determine the amount of glucose (mmol/L) present
in the well. Read the test strip 1 minute after removing it from the well.

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 BIOL-101.602 GENERAL BIOLOGY I
MATH AND SCIENCES DEPARTMENT
Fall 2023

Lab Report

10.Record the amount of glucose present in Data Table 1 under “Glucose Concentration”.

11.Repeat steps 13 through 16 for the remaining 4 wells (B, C, and D).

Cleanup:

 Discard the used glucose strips in the trash when complete.

 Keep all solutions and other materials on your lab bench.

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 BIOL-101.602 GENERAL BIOLOGY I
MATH AND SCIENCES DEPARTMENT
Fall 2023

Lab Report

B. Enzymes and pH

1. Label 4 empty wells in the 24-well plate “1, 2, 3, and 4”.

2. Use the pH 3.5, pH 5.0, pH 6.8, and pH 11.5 pipets. Place each pipet bulb side down (tip side up)
in empty wells of the 24-well plate. See Figure below.

3. As described in Data Table 2, use the pH pipets, and labeled pipets, to add the following solutions
into the following wells:

a) In Well 1: add 3 drops of pH 3.5 buffer, 2 drops of lactase, and then 2 drops of milk

b) In Well 2: add 3 drops of pH 5.0 buffer, 2 drops of lactase, and then 2 drops of milk

c) In Well 3: add 3 drops of pH 6.8 buffer, 2 drops of lactase, and then 2 drops of milk

d) In Well 4: add 3 drops of pH 11.5 buffer, 2 drops of lactase, and then 2 drops of milk

Note: Ensure that pipets are only used to transfer solutions from their designated cups.

4. Allow each of the reactions to occur for 10 minutes.

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 BIOL-101.602 GENERAL BIOLOGY I
MATH AND SCIENCES DEPARTMENT
Fall 2023

Lab Report

5. After 10 minutes, pick up a fresh glucose strip and insert it into the reaction in Well 1. Keep the
strip in the solution for 5 seconds.

6. After 5 seconds, remove the strip from the well and place it onto the paper towel.

7. Use the Glucose Color Chart to determine the amount of glucose present (mmol/L) in the well.
Read the test strip 1 minute after removing it from the well.

8. Record the amount of glucose present in Data Table 2 under “Glucose Concentration”.

9. Repeat steps for the remaining three wells (2, 3, and 4).

Cleanup:

 Discard the used glucose strips in trash when complete.

 Use the dish soap and brush to wash all well plates and glassware.

 Keep all remaining solutions

GOOD LUCK!

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