CHY 47 

(BSN 1C) 
ASSIGNMENT 5 

Answer the following as directed. 

1. Fill in the table below. To which class of enzymes (Column B) does each of the following enzymes in
Column A belong? Write your answer on the space provided before each number. (10 pts.)   

          Column A Column B 

     D     a. Pyruvate kinase A.  ligase 
     D     b. Alanine transaminase B.  hydrolase 
     G     c. Triose phosphate isomerase C.  oxidoreductase 
     G     d. Phosphoglycerate mutase D.   transferase 
     B     e. Lactase E.  lyase 
    D     f. Phosphofructokinase F.  transferase 
     B     g. Lipase G.  isomerase 
     E     h. Acetoacetate decarboxylase 
     C     i. Succinate dehydrogenase 
     C      j. catalase 

2. Draw and label a reaction coordinate diagram for an uncatalyzed reaction with substrate, S
and product, P and the same reaction coordinate diagram of catalyzed reaction by an enzyme, E.  (5 
points)

Figure 1. Reaction Coordinate Diagram for an Figure 2. Reaction Coordinate Diagram for a
Uncatalyzed Reaction Catalyzed Reaction
3. List and explain the factors affecting enzyme activity.  (10 pts.)

Enzyme activity can be affected by a variety of factors, such as temperature, pH, enzyme
concentration, substrate concentration and inhibitor.

 Effect of Temperature
Raising temperature speeds up a reaction, and lowering temperature slows down a
reaction. As the temperature increases, more of the reacting molecules have enough kinetic
energy to undergo the reaction. Since the temperature of an enzymatically catalyzed
reaction increases, so does the rate of the reaction. However, if the temperature of an
enzyme – catalyzed reaction is raised still further, an optimum temperature is reached.
Further elevation of the temperature results in a decrease in reaction velocity as a result of
temperature-induced denaturation of the enzyme leading to derangement in the native
(tertiary) structure of the protein and active site.
Low temperatures decrease the system's kinetic energy and hence the pace of reaction.
As the temperature falls below the optimum level, enzyme activity decreases. In contrast to
high temperatures, low temperatures do not always result in permanent enzyme
denaturation, and enzyme function can be restored if the temperature returns to the
optimum range. Because enzymes are often found in aqueous solutions, a fall in
temperature alters the nature of their interaction with water, lowering solubility and
causing the enzyme to unfold - ultimately inactivating the enzyme. However, when the
temperature goes below the melting point of water (0°C or 32°F), ice crystals develop, which
can irreversibly damage the proteins. When frozen enzymes are thawed, the same action is
observed. By reducing freeze-thaw cycles, freezing or thawing time, and adding additives
like sucrose or glycerol to the protein solution, freeze-thaw damage can be minimized.

 Effect of pH
Proteins, like enzymes, have electrical charges owing to the sequence of their amino
acid residues since they are made up of a chain of amino acids. The majority of the amino
acids in the chain serve as the foundation for the intramolecular interactions that give the
enzyme its three-dimensional shape. Few others serve as functional residues at the active
site of the enzyme. Overall, the amino acids dictate substrate selectivity and limit enzyme
activity to a narrow pH range. Most enzymes work best in slightly acidic or basic
environments. However, a few enzymes are native to extremely acidic or basic
environments and, as a result, are most active in these pH ranges. As a result, a change in
pH, whether acidic or basic, affects the ionization of amino acid residues, resulting in
changes in the three-dimensional structure of the enzyme. The change in enzyme shape
alters the enzyme's interaction with its substrate, decreasing its activity. Another impact of
pH change is an increase in the catalytic capacity of the enzyme. Changes in pH can impede
or inhibit catalytic activity in acid-base and covalent catalysis processes. It can denature the
enzyme, ruin its three-dimensional structure, and render it permanently inactive in severe
circumstances.

 Concentration of Enzyme
If the amount of substrate present remains constant but the enzyme concentration
increases, the reaction rate increases proportionally because more substrate molecules may
be accommodated in the same period of time. Once all of the substrate has been bound, the
 process will no longer be able to accelerate since there will be nothing for further enzymes
to bind to.

 Concentration of Substrate
The rate of an enzymatic reaction increases as the substrate concentration increases in
the presence of a given quantity of enzyme until a limiting rate is achieved; beyond that,
additional increases in the substrate concentration cause no meaningful change in the
reaction rate. At this point, there is so much substrate that it has bonded to almost all of the
enzyme active sites. To put it another way, the enzyme molecules have been saturated with
substrate. The excess substrate molecules will not be able to react until the substrate that is
already attached to the enzymes has reacted and been released.

 Inhibitor
Many enzymes rely on non-substrate and non-enzyme substances to control or begin
catalysis. Certain enzymes, for example, rely on metal ions or cofactors to create catalytic
activity. Many enzymes, such as allosteric enzymes, rely on effectors to activate their
catalytic activity and promote or prevent their subsequent binding to substrates. Similarly,
inhibitors may attach to the enzyme or its substrate to inhibit ongoing enzymatic activity and
prevent further catalytic events. Inhibitors are chemicals that slow down the rate of enzyme
activity by interacting with the enzyme and interfering with the formation of the enzyme-
substrate complex. When inhibitors create strong bonds to the enzyme's functional group,
the effect on enzyme activity is irreversible, rendering the enzyme irreversibly inactive.
Reversible inhibitors, as opposed to irreversible inhibitors, only render enzymes inactive
while attached to the enzyme. Competitive inhibitors compete with substrates for binding to
the enzyme functional group residues at the catalytic sites. Other kinds of inhibitors bind to
the non-substrate binding allosteric site rather than the catalytic site. It is non-competitive if
an inhibitor attaches to the enzyme concurrently with the enzyme-substrate interaction . An
inhibitor is uncompetitive if it binds solely to a substrate-occupied enzyme.

4. Explain the different theories proposed for mechanism of enzyme-substrate complex formation. 
Show illustrations where possible.  (6pts.)

Two theories have been proposed to explain the mechanism and enzyme action. They are
Fischer's Lock and key theory and Koshland's induced fit theory.

 Fischer’s Lock and Key Theory

Emil Fischer proposed the Lock and Key analogy in 1894 to describe the specific activity
of an enzyme with a single substrate. In this analogy, the lock represents the enzyme and
the key represents the substrate. Only the right sized key (substrate) fits into the lock's
(enzyme's) key hole (active site) (Figure 3).

To elaborate, the Lock and Key Model argues that the active site and substrate complement
each other in such a way that the substrate fits precisely into the binding site. The active site
of the enzyme has a set, rigid geometrical shape in this model. At such a location, only
substrates with comparable geometry may be accommodated. This approach, however,
 does not apply to all enzymes. This model fails to account for protein conformational
changes in order to accommodate a substrate molecule.

Figure 3. Lock and Key Analogy

 Koshland’s Induced Fit Theory

Not all experimental evidence can be effectively described by the lock and key theory's
so-called rigid enzyme model. As a result, Koshland suggested a modification known as the
induced-fit theory.

In induced-fit model of enzyme action, the enzyme undergoes a conformational change

upon binding to substrate. The shape of the active site becomes complementary to the
shape of the substrate only after the substrate binds to the enzyme. It is assumes that the
active region of the enzyme is a more flexible pocket whose conformation changes to accept
the substrate molecule. It allows for minor modifications in the structure or geometry of an
enzyme's active site in order to accept a substrate. This means that when an enzyme binds
its substrate, it slightly alters shape, resulting in an even tighter fit. This adjustment of the
enzyme to snugly fit the substrate is called induced fit.

Figure 4 depicts an illustration of the induced fit model of enzyme catalysis. When a
substrate attaches to the active site, the active site changes shape slightly, tightening its grip
on the substrate and prepares to catalyze the reaction. Following the reaction, the products
are released from the active site and disperse away.

Figure 4. Induced Fit Model

 This is a more comprehensive explanation for an enzyme's active-site characteristics since it
incorporates the specificity of the lock-and-key model as well as the flexibility of the enzyme
protein.

SOURCES:
 https://conductscience.com/factors-that-affect-enzyme-activity/
 https://www.khanacademy.org/science/ap-biology/cellular-energetics/enzyme-structure-
and-catalysis/a/enzymes-and-the-active-site
 https://saylordotorg.github.io/text_the-basics-of-general-organic-and-biological-
chemistry/s21-06-enzyme-action.html
 https://www.futurelearn.com/info/courses/the-biology-of-bugs-brains-and-
beasts/0/steps/68820
 https://classroom.google.com/u/0/c/MzgxNjI0MTYwMDY4/m/NDA2NDcyODkwMjY0/detail
s
 https://classroom.google.com/u/0/c/MzgxNjI0MTYwMDY4/m/NDA2NTA3NDQ3NDI2/detail
s