Name: Andreah B.

Baylon Course & Year: BSBiotech- 2

Offering No.: R173 Class Schedule: TF 2:30 - 4:00
Lesson 6
ENZYMES

1. Lock-and-key model shows that the active site of the enzyme is rigid and perfectly

complementary to the substrate, like a key to a lock. On the other hand, induced-fit

model tells that the enzyme is flexible and undergoes conformational changes when

binding to a substrate – the active site adapts to the substrate’s shape.

2. Proteolytic enzymes are synthesized as inactive zymogens to prevent unwanted protein

degradation or protein digestion in the cell as inactive zymogens are harmless until

activate. Also to regulate its enzymatic activity, within activation it can be controlled

allowing control to its proteolytic activity.

3. Not all enzymes display kinetics that obey Michaelis-Menten equation such as allosteric

enzymes, multi-substrate/product enzyme, irreversibly-inhibited enzymes, and substrate-

inhibited enzymes.
4. When its graph on rate of activity and substrate concentration is sigmoidal – S-shaped

curve on its kinetics and not linear.

5. It is good as it allows precise control over the activity like regulation of metabolic

pathway and protection against harmful substance

6. Lessons that may aid in research and development against AIDS are Enzymes that is

involved in the life cycle on HIV, having a possible effect on reverse transcription,

inhibitions, protease-cleavage protein. Additional topics are enzymes involved in immune

response in our body, for replication and enzymes that is involved in DNA transcription

and translation.

7. Irreversible inhibitor could be bound by covalent interactions as it is stronger and requires

significant energy to separate and break as irreversible inhibitor permanently inactivates

an enzyme.

8. No, since noncompetitive inhibitor binds to the enzyme’s allosteric sites rather than the

active site. Allosteric site is far from the active site.

9. The kinetic data from an enzyme catalyzed reaction is given below.

a. (5 points) Using the data in the table below, graph the correct Lineweaver-

Burk (double reciprocal) plot. Label the x and y axis with the correct units.

1 1
[𝑆] VO 𝑉
[S] (µmoles/L) (L/µmoles) (minutes/µmole)
(µmoles/minute)
(x-intercept) (y-intercept)
0.1 10 3.33 0.3003003003
0.2 5 5.00 0.2
0.5 2 7.14 0.1400560224
0.8 1.25 8.00 0.1250
1.0 1 8.3 0.1204819277
2.0 0.5 9.09 0.1100110011
 b. (5 points) From the Lineweaver-Burk plot, determine the Km and Vmax

of the enzyme.

Show your work for partial credit.

(minutes/µmole)

µmoles-1
 c. (5 points) On the same graph, plot a dashed line that would show graphically

how the addition of a non-competitive inhibitor would affect the Vmax and Km.

(Assume that the values for non-competitive inhibitor is provided and plotted)

d. (5 points) Briefly describe in words what is happening to Km and Vmax in

question c.

In addition of Noncompetitive Inhibitor, the Vmax decreased its value and the Km

remain unchanged.
Assessment

1. The kinetic data from an enzyme catalyzed reaction is given below.

1 1
[𝑆] VO 𝑉
[S] (µmoles/L) (L/µmoles) (minutes/µmole)
(µmoles/minute)
(x-intercept) (y-intercept)
1.5 0.666666667 0.21 4.761904762
2.0 0.5 0.24 4.166666667
3.0 0.3333333333 0.28 3.571428571
4.0 0.25 0.33 3.03030303
8.0 0.125 0.40 2.5
16.0 0.0625 0.45 2.222222222

(10 points) From this information determine the Km’s and Vmax’s for the enzyme by plotting

the data on a Lineweaver Burk (double reciprocal) plot and calculating the values for Km and

Vmax. Label X-axis and Y-axis with correct title and units.

1
Vmax =
1.9978 = 0.5005506057 minutes/µmole

Km = (m)(Vmax) = (4.2663)( 0.5005506057) = 2.135499049 µmoles-1

b) (5 points) On the same graph, plot a dashed line that would show graphically how the

addition of a non-competitive inhibitor would effect the Vmax and Km.

(Assume that the values for non-competitive inhibitor is provided and plotted)

c) (5 points) Briefly describe in words (two sentences would be enough) how the

addition of a competitive inhibitor would effect the Vmax and Km.

In the addition of competitive inhibitor, Km will increase. The Vmax while remain

unchanged.

2. List six general ways in which enzyme activity is controlled.

a. Allosteric binding sites

b. By Covalent Modification

c. Induction and Repression of Enzyme Synthesis

d. Zymogen Cleavage

e. Location within the cell (Compartmentalization)

f. Environmental Factors (e.g. pH, temperature)

3. The following graphs show the temperature and pH dependencies of four

enzymes, A, B, X, and Y. Problems 3 through 6 refer to these graphs

Enzymes X and Y in the figure are both protein-digesting enzymes found in

humans. Where would they most likely be at work?

A] X is found in the mouth, Y in the small intestine. B]

X in the small intestine, Y in the mouth.

C] X in the stomach, Y in the small intestine. D]

X in the small intestine, Y in the stomach.

Answer: C] X in the stomach, Y in the small intestine.

4. Which statement is true concerning enzymes X and Y?

A] They could not possibly be at work in the same part of the body at the same time. B] They

have different temperature ranges at which they work best.

C] At a pH of 4.5, enzyme X works slower than enzyme Y.

D] At their appropriate pH ranges, both enzymes work equally fast.

ANSWER: B] They have different temperature ranges at which they work

best.

5. What conclusion may be drawn concerning enzymes A and B?

A] Neither enzyme is likely to be a human enzyme. B]

Enzyme A is more likely to be a human enzyme. C]

Enzyme B is more likely to be a human enzyme. D]

Both enzymes are likely to be human enzymes.

ANSWER: B] Enzyme A is more likely to be a human enzyme.

6. At which temperatures might enzymes A and B both work?

A] Above 40°C

B] Below 50°C

C] Above 50°C and below 40°C D]

Between 40° and 50°C

ANSWER: D] Between 40° and 50°C