Enzyme Lab Outline

1. Introduction
1. Basic background
1. Enzymes are catalysts that speed up chemical reactions 
1. Not part of chemical makeup, reused
2. Most reactions would take too long without enzymes 
3. Enzymes lower activation energy
2. Enzymes are proteins
1. 3-d structure has one or more polypeptide chains that fold
and create active site
2. Substrate attaches to active site
3. Enzymatic activity of sucrase
1. Produced by yeast
2. Sucrose is too large to pass through membrane
1. Yeast allows sucrase to pass through which catalyzes
hydrolysis of sucrose into glucose and fructose
b. Benedict’s reaction
1. Indicates presence of reducing sugars
2. Turns from blue to green, yellow, orange or red if positive
1. Blue-0
2. Green-1
3. Yellow-2
4. Orange-3
5. Red-4
ii. Changes in temperature, alterations in pH, addition of certain ions or
particles, and presenc of inhibitors affect enzymes active site
a. Temperature
1. Rate of reaction increases with temperature until maximum
2. Above 40°C most enzymes become denatured
b. pH
1. Optimum pH is where enzymes are most active
2. Extremes of pH denature enzymes
c. Denaturation on enzyme activity
1. When a substrate fits into the active site, enzyme-substrate
complex is formed
2. If altered, enzyme cannot catalyze reaction
d. Substrate concentration
1. If the amount of substrate is increased while the amount of
enzyme is kept constant, the velocity is increased until
maximum is reached
ii. Purpose of lab is to investigate how changes in pH, temperature,
substrate concentration, and enzyme denaturation affect the enzymatic
activity of sucrase
iii. Hypothesis
b. Materials and Methods
i. From lab manual
 b. Results
Effect of Temperature on Enzyme Activity
Tube Temperature Color Observations Color Score
# °C

1 0°C

2 25°C

3 37°C

4 55°C

5 100°C

Effect of pH on Enzyme Activity

Tube p Color Observations Color Score
# H

1 3

2 5

3 7

4 9

5 11

Effect of Denaturation on Enzyme Activity

Tube Treatments Color Observations Color Score
#

1 boiled sucrase/ boiled sucrose

2 boiled sucrase/ untreated sucrose

3 untreated sucrase/ boiled sucrose

4 untreated sucrase/ untreated

sucrose

Effect of Substrate Concentration on Enzyme Activity

Tube Substrate Color Color Score
# Concentration  Observations
 1 0%

2 10%

3 27%

4 50%

5 75%

6 100%
Graphs
4. Discussion
a. Temperature
i. Rate of reaction at different temperatures
ii. Temperature at which the greatest amount of product was produced by
the enzyme-catalyzed reaction
iii. How product produced per minute relates to the rate of enzyme-catalyzed
reaction
iv. Temperature at which the enzyme-catalyzed was greatest
b. pH
i. pH at which the greatest amount of product was produced by the enzyme-
catalyzed reaction
ii. How product produced per minute relates to the rate of enzyme-catalyzed
reaction
iii. pH at which the enzyme-catalyzed was greatest
b. Denaturation
i. Condition at which the greatest amount of product was produced by the
enzyme-catalyzed reaction
ii. Condition at which the least amount of product was produced by the
enzyme-catalyzed reaction
b. Substrate Concentration
i. Condition at which the greatest amount of product was produced by the
enzyme-catalyzed reaction
ii. Condition at which the least amount of product was produced by the
enzyme-catalyzed reaction
iii. Support of maximum velocity 
b. Sources of error
c. What was learned