Title:

“The Effect of Varying Temperature on

Enzymatic Activity of Aspergillus oryzae and
Bacillus licheniformis During Starch´s
Hydrolysis”

Author: Cynthia Hernandez

Panther ID: 6164473
Biology Lab I
Section: U11
Lab Partners:
Brittani Hjerpe
Samuel Solano
Gabriela Puentes.
Abstract:

The main objective of this experiment was to test the effect of temperature on the

enzymatic activity during the process of breaking down starch through hydrolysis. In order to do

it, two different sources of amylases (a type of enzyme) were used, Bacillus licheniformis a

bacterial amylase, which came from a sample of the bacterium; and Aspergillus oryzae a fungal

amylase which came from a sample of the fungus. The experiment was set with 4 different

temperatures (0 C, 25 C, 55 C, 85 C) and 5 time intervals (0, 2, 4, 6, 8 and 10 minutes) in order

to test how the temperature affected the enzymatic activity during the time intervals, and found

out at which time interval occurred the 100% of hydrolysis. In addition, 0 minutes was the

control group established to have certainty that the enzymes were the only responsible for

breaking down the starch.

In order of detect the presence of starch, iodine was used as an indicator due to the reaction

between iodine and starch turn the reaction black, and in case of starch’s absence, the reaction

became yellow. This color’s change was the indicator of enzymatic activity. If the enzymes were

working at their optimum temperature, which is 55 C for Bacillus licheniformi and 25 C for

Aspergillus oryzae, the reaction should turn yellow because the starch broke down. If the

difference between the optimum temperature and the one present in the reaction is too wide, the

reaction could remain black because the enzymes were not working and the starch was present.

However, if the difference between optimum and present temperature of the reaction is narrow

then the reaction could turn brown because the enzymes are working but not at their optimum

temperature, so some starch was still present in the reaction.

Introduction:

During a process called catalysis, enzymes, which are constituted by proteins, take the

role of enhancing chemical reactions. The first step in this process occurs when the substrates

(molecules that undergo the reaction) binds to the active side of the enzyme to form a product (a

different molecule that the one that undergo the reaction). This binding occurs in the enzyme-

substrate complex, this complex is formed when the substrate and the enzyme bind each other.

(Alberte J., Pitzer T., Calero K., 2012)

The first model to explain this process was called Lock and Key Model. It received this

name because scientists believe that the active site on the enzyme (lock) were unique and just

allowed binding to a specific substrate (key) that match its shape. However, science advances

continually and a second model called Induce Fit Model appeared. This model explained that the

active side is flexible and it could adjust itself to match the shape of the substrate allowing the

binding between the substrate and the active site (Raven, 2008; Ringe & Petsko, 2008;

Whitehurst & Van Oort, 2009).

On the other hand, enzymes lower the amount of energy that a chemical reaction required

to take place. That is why enzymes are considered a catalyst. They lower the amount of energy

that exists between the reactants and the transitional state. A good analogy that could be applied

to this statement it would be when a person took a shortcut to get his or her destination faster.

The making of that shortcut is one of the functions of the enzymes. Another interesting

fact about this process is that the enzymes are not consumed or altered during or after the

enzymatic activity. The body recycles them. In addition, enzymes do not affect the reactions’

equilibrium (Garcia, Gao., Karplus, Truhlar, 2004; Raven, Ringe & Petsko, 2008; Whitehurst &

Van Oort, 2009; Alberte et al., 2012).

 However, enzymatic activity is affected by different elements such as temperature,

because enzymes have an optimum temperature point. If the temperature of the reaction exceeds

this optimum point will cause a denaturation of the enzymes and a change in shape implies a

change in their function so the enzymes will not be able of work properly. Moreover, if the

temperature of the reaction is too low, the substrates will not be able of bind to the active site

because the active site will not be flexible enough to fit with the shape of the substrate, also the

number of interactions between substrate and active site is going to decrease. (Urry, Cain,

Minorsky Wasserman and Pearson Reece, 2016)

Others elements that also affect enzymatic activity are PH (ranges from 6 to 8), substrate

concentration because if the substrate amount increases and the enzyme amount remains the

same, the velocity of reaction it is going to increase until the maximum point is reached, and it is

going to remain the same after that, even though more substrate is added. Allosteric inhibitors

and activators could also affect the enzymatic activity. Inhibitors (competitive and no

competitive) decrease the activity because they avoid that the substrate binds to the active site;

and allosteric activators enhance enzyme activity because they bind to allosteric site keeping the

enzyme in an active configuration. The last element is called cofactor. Cofactors could be a

prosthetic group, or metal ion activator or coenzymes, which the enzymes need for the catalytic

activity (Harisha, 2006; Raven Johnson G. B., Mason K. A., Losos J. B., Singer S. S. 2008;

Whitehurst & Van Oort, 2009).

Enzymes are used in the modern world in different areas such as medicine, brewing

process, and baking. Enzymes are also present in signal transduction to generate muscle

contraction. Moreover, the organisms used them in metabolic process to obtain energy (Alberte

J., Pitzer T., Calero K. 2012).

 The type of enzyme amylases is recognized because it can breakdown starch (polymer) in

smaller monomers, which the organisms can absorb easily through hydrolysis (chemical

breakdown of a compound due to reaction with water) (Whitehurst & Van Oort, 2009; Alberte et

al., 2012).

This specific ability of amylases to be able to breakdown starch will be analyzed in this lab

report. The objective is to test the effect of the temperature in two different source of amylases,

Bacillus licheniformis (Bacterial amylases) and Aspergillus oryzae (fungus amylases) when they

are acting in a solution that contains starch. The indicator of starch that will be used is iodine

because if starch is present the reaction with iodine is going to turn black, and if is not present

then the reaction is going to turn yellow. Thus, if the enzyme does not work, the solution will

remain black, if the enzyme works, the solution will become yellow, and if the enzyme works but

not at its full potential because the temperature was not the optimal one, the reaction will have a

color´s change from black to brown.

Therefore, during the process of breaking down starch using amylases, if the temperature

increases or decrease drastically reminding below or above of the optimum point of the specific

enzyme (amylase). Then the enzymatic activity is going to reduce or stop because the

temperature affect the enzymatic activity producing a denaturation of the protein that compose

the enzyme, if it is too hot , or if it is too cold there is no not going to be enough interaction

between the substrate and the active site for them to work.

There are two possible explanations for the success or failure of this experiment. If the

experiment fails is probably because there is no correlation between the temperature´s change

and the enzymatic activity; therefore, color change in the sample is not going to occur.
 On the other hand, if the experiment succeeds is because there is correlation between the

temperature´s change and the enzymatic activity; therefore, a color change in the samples is

going to occur.
Methods:
Procedure:

The experiment’s objective was to determine the optimal temperature for Aspergillus

oryzae (fungus amylases) and Bacillus licheniformis (bacterial amylases). The students tested

how the temperature affected the amylases function at breaking down starch. In order to do it, the

students used two napkins, two spot plates (one for the bacteria amylases and another one for

fungus amylases), sixteen test tubes (eight for the bacteria amylases and another eight for fungus

amylases), and 5ml of 1.5% starch solution.

First, the students marked on the top of the napkin the following temperatures (0 C, 25 C,

55 C, 85 C), and on the side the time (0, 2, 4, 6, 8, 10 min). The following step was to label the

test tubes with the different temperatures mentioned above (four tubes each one), and the letters

FA or BA to differential between the amylase’s source (fungal or bacterial), (eight tubes each

one). In addition, the students added and S to four of the tubes per each amylases sources and the

team number to avoid confusion.

In the next step, the students added 5 ml of 1.5% of starch solution into each of the test

tubes labeled with S. After this, the students added 1 ml of fungus amylase into each of the four

test tubes that do not contain starch, and 1 ml of bacterial amylases into 4 of the test tubes that

not contain starch. The tubes were placed in their correspondent temperature and the students

allowed them 5 minutes of equilibration time.

Two drops of iodine was then added to the plate at a 0 minute interval. When the

equilibration process was finished, the students transferred a few drops of the starch solution to

its correspondent temperatures at 0 minutes, using a different pipette for each temperature. In the

following step, the starch solution was added to the tubes containing amylase and the timer was

set for 2 minutes.

 Two Drops of iodine were added to each of the 0 minutes´ row before transferring each of

the amylases to its correspondent spot place. The Amylases were transferred every two minutes

with its correspondent pipette to its corresponding temperature and time. The students recorded

the color change observations after being sure that they mixed well the reactants. After an

interval of ten minutes, the students were able to identify the temperature and time at which

100% of hydrolysis occurred. Finally, using the color´ scheme the data was converted from

qualitative (color) to quantitative (numerical)

Results:

Prediction:

Data Table #1. Bacterial Amylase Predictions

Temperature C Expected Result Reasoning

0 No change in color No enzymatic activity

25 Little change in color Some enzymatic

activity
55 Color´s change Optimal enzymatic

activity
85 No color change No enzymatic activity

Data Table #1.2 Fungal Amylase Predictions

Temperature C Expected Result Reasoning

0 No enzymatic No enzymatic activity

activity
25 Optimal enzymatic Optimal enzymatic

activity activity
55 Little change in Some enzymatic activity

color
85 No color change No enzymatic activity

Team Result:

Data Table # 2. Bacterial Amylase (teamwork)

Temp C/ 0 0 25 25 55 55 85 85

Time min
Color # Color # Color # Color #
0 Black 5 Black 5 Black 5 Black 5
2 Dark 4 Light 3 Dark Yellow 2 Black 5
 Brown Brown
4 Light 3 Light 3 Dark Yellow 2 Black 5

Brown Brown
6 Light 3 Light 3 Dark Yellow 2 Black 5

Brown Brown
8 Light 3 Light 3 Dark Yellow 2 Black 5

Brown Brown
10 Light 3 Light 3 Dark Yellow 2 Black 5

Brown Brown
TIME TO 100% 2 minutes

HYDROLYSIS

AT OPTIMAL

TEMP
OPTIMAL 55C

TEMP.

Figure #1. Starch hydrolysis. Yellow means not starch presences and black means starch

presences.
 The tables above showed the result of the experiment regarding bacterial amylase. As the

reader could observe, 0 minutes is the control group because it remained constant (5), without

any color’s change from black to yellow when iodine was added, because enzyme amylases were

not added to this group.

On the other hand, the group with 85 C temperatures did not suffer any color changes

either, because the temperature was above from the optimal point for this enzyme, that is 55C.

Therefore, the high temperature produced a denaturation of the enzyme. This caused a change on

the shape of the enzyme and in the end, a change in its function too, producing no effect when it

reacted with the starch.

When the solution with starch and amylases was exposed to 0 C temperatures, the color

changed from black to light brown when iodine was added. The reason was that the temperature

was too low and caused a reduction of the interaction between subtracts and the active site.

However, when the solution was exposed to 25C, it showed a color change from black to

light brown due to the fact that the temperature was not the optimal one, but produced more

interaction between substrate and active side than the solution exposed to 0 C. In addition, this
temperature did not affect the shape of the enzyme, as occurred when the substance was expose

to a temperature of 85 C.

Finally, the optimal point of enzymatic activity was reached at 55 C after 2 minutes of

exposure of the solution of starch and amylase to that temperature. Here, it was the spot plate

which suffered the biggest change in color, going from a black or dark blue (indicator of starch’s

presence) to a dark yellow (indicator of no starch’s presence) because the enzyme broke down

the starch bonds

Data Table 3. Fungal Amylase (Team work):

Temp C/ 0 0 25 25 55 55 85 85

Time min
Color # Color # Color # Color #
0 Black 5 Black 5 Black 5 Black 5
2 Dark 4 Dark 4 Dark 2 Black 5

Brown Brown Yellow

4 Dark 4 Dark 4 Dark 2 Black 5

Brown Brown Yellow

6 Dark 4 Light 3 Dark 2 Black 5

Brown Brown Yellow

8 Dark 4 Light 3 Dark 2 Black 5

Brown Brown Yellow

10 Dark 4 Light 3 Dark 2 Black 5

Brown Brown Yellow

TIME TO 2 minutes

100%

HYDROLYSIS

AT OPTIMAL
TEMP
OPTIMAL 55 C

TEMP.

Figure #2. Starch hydrolysis. Yellow means no starch presences and black means starch

presences

The table above showed the result for the experiment regarding fungal amylase. As the

reader could observe 0 minutes is the control group because it remains constant (5), without any

color’s change from black to yellow when iodine was added, due to fact that enzyme amylases

were not added to this group.

On the other hand, the group with 85 C temperatures did not suffer any color changes

neither, because the temperature was above up to the optimal point for this enzyme that

apparently is 55C. Thus, the high temperature produced a denaturation of the enzyme. This

caused a change of shape of the enzyme ending in a change in its function too, producing no

effect when it reacted with the starch.

 In addition, when the solution with starch and amylases was exposed to 0 C temperatures,

the color changed from black to dark brown when iodine was added. The reason was that the

temperature was too low and caused a reduction in the interaction between the subtracts and the

active site.

However, when the solution test was exposed to 25 C, had a color change from black to

light brown due to the fact that the temperature was not the optimal one, but produced more

interaction between substrate and active side than the solution exposed to 0 C. Furthermore, it

did not affect the shape of the enzyme as occurred when the substance was exposed to a

temperature of 85C.

Finally, the optimal point of enzymatic activity was reached at 55 C after 2 minutes of

exposure to the solution of starch and amylase at this temperature. Here, it was the spot plate that

suffer the biggest change in color, going from a black or dark blue (indicator of starch’s

presence) to a dark yellow (indicator of no starch’s presence) because the enzyme broke down

the starch bonds.

Class Results:

Data Table 4. Bacterial Amylase. This Table shows the color´s change produced by varying

the temperatures in a reaction that contain starch and bacterial amylases.

Bacterial Amylase
0 25 55 85

Temp (°C)

Time (min)
Group 1 5.00 5.00 5.00 5.00
Group 2 5.00 5.00 5.00 5.00
Group 3 5.00 5.00 5.00 5.00
Group 4 5.00 5.00 5.00 5.00
Group 5 5.00 5.00 5.00 5.00
Group 6 5.00 5.00 5.00 5.00
0 minutes 5.00±0.00 5.00±0.00 5.00±0.00 5.00±0.00

Mean ± SD
Group 1 4.50 4.50 2.00 5.00
Group 2 4.00 3.00 2.00 5.00
Group 3 4.00 3.00 2.00 5.00
Group 4 4.00 3.00 2.00 5.00
Group 5 4.00 3.00 2.00 5.00
Group 6 5.00 4.00 2.00 5.00
2 minutes 4.25±0.42 3.42±0.66 2.00±0.00 5.00±0.00

Mean ± SD
Group 1 4.00 4.00 2.50 5.00
Group 2 4.00 3.00 2.00 5.00
Group 3 4.00 3.00 1.00 5.00
Group 4 4.00 3.00 2.00 5.00
Group 5 3.00 3.00 2.00 5.00
Group 6 5.00 4.00 2.00 5.00
4 minutes 4.00±0.63 3.33±0.52 1.92±0.49 5.00±0.00

Mean ± SD
Group 1 3.50 4.00 2.50 5.00
Group 2 4.00 3.00 2.00 5.00
Group 3 4.00 3.00 2.00 5.00
Group 4 4.00 3.00 2.00 5.00
Group 5 3.00 3.00 2.00 5.00
Group 6 4.00 4.00 2.00 5.00
6 minutes 3.75±0.42 3.33±0.52 2.08±0.20 5.00±0.00

Mean ± SD
Group 1 3.50 4.00 2.50 5.00
Group 2 4.00 3.00 2.00 5.00
Group 3 4.00 3.00 2.00 5.00
Group 4 4.00 3.00 2.00 5.00
Group 5 3.00 3.00 2.00 5.00
Group 6 4.00 4.00 1.00 5.00
8 minutes 3.75±0.42 3.33±0.52 1.92±0.49 5.00±0.00

Mean ± SD
Group 1 3.50 4.00 2.50 5.00
Group 2 3.00 2.00 1.00 5.00
Group 3 4.00 3.00 2.00 5.00
Group 4 4.00 3.00 2.00 5.00
Group 5 3.00 3.00 2.00 5.00
Group 6 4.00 3.00 1.00 5.00
10 mintues 3.58±0.49 3.00±0.63 1.75±0.61 5.00±0.00

Mean ± SD
OPTIMAL TEMP. Optimal

Temperature
TIME TO 100% 10 min

HYDROLYSIS AT

OPTIMAL TEMP.

Graph 2: This graph shows that the optimal temperature of bacterial amylase is at 55 C,

and the 100% hydrolysis was reached at the 10 minutes interval.

Bacterial Amylase at 10 min

6

5
Color Change

0
0 25 55 85

Temperature (°C)
 The tables above showed the results for the experiments regarding bacterial amylase. As

the reader could observe, 0 minutes is the control group because it remains constant (5), without

any color’s change from black to yellow when iodine was added, due to fact that enzyme

amylases were not added to this group.

On the other hand, the group with 85 C temperatures did not suffer any color change

neither, because the temperature was above of the optimal point for this enzyme that is 55C.

Thus, the high temperature produced a denaturation of the enzyme. This causes a change on the

shape of the enzyme and a change in its function too, producing no effect when it reacted with

the starch.

When the solution with starch and amylases was exposed to 0 C temperatures, the color

changed from black to dark brown when iodine was added. The reason was that the temperature

was too low and caused a reduction of the interaction between subtracts and the active site.

However, when the solution was exposed to 25C, it had a color change from black to

light brown due to the temperature was not the optimal one, but produced more interaction

between substrate and active side than the solution exposed to 0 C. In addition, this temperature

did not affect the shape of the enzyme, as occurred when the substance was exposed to a

temperature of 85C.

Finally, the optimal point of enzymatic activity was reached at 55 C after 10 minutes of

exposure of the solution of starch and amylase to that particular temperature. This was the spot

plate that suffered the biggest change in color, going from a black or dark blue (indicator of

starch’s presence) to a light yellow (indicator of no starch’s presence) because the enzyme broke

down the starch bonds.

Data Table 5. Fungal Amylase. This Table shows the color´s change produced by varying

the temperatures in a reaction that contain starch and fungal amylases.

Fungal Amylase
0 25 55 85

Temp (°C)

Time (min)
Group 1 5.00 5.00 5.00 5.00
Group 2 5.00 5.00 5.00 5.00
Group 3 5.00 5.00 5.00 5.00
Group 4 5.00 5.00 5.00 5.00
Group 5 5.00 5.00 5.00 5.00
Group 6 5.00 5.00 5.00 5.00
0 minutes 5.00±0.00 5.00±0.00 5.00±0.00 5.00±0.00

Mean ± SD
Group 1 4.50 4.00 3.00 5.00
Group 2 5.00 4.00 3.00 5.00
Group 3 5.00 5.00 4.00 5.00
Group 4 4.00 4.00 3.00 5.00
Group 5 4.00 4.00 2.00 5.00
Group 6 5.00 4.00 3.00 5.00
2 minutes 4.58±0.49 4.17±0.41 3.00±0.63 5.00±0.00

Mean ± SD
Group 1 4.50 3.50 3.00 5.00
Group 2 5.00 4.00 3.00 5.00
Group 3 5.00 5.00 4.00 5.00
Group 4 4.00 4.00 3.00 5.00
Group 5 4.00 4.00 2.00 5.00
Group 6 4.00 4.00 3.00 5.00
4 minutes 4.42±0.49 4.08±0.49 3.00±0.63 5.00±0.00

Mean ± SD
Group 1 4.50 3.50 3.00 5.00
Group 2 4.00 4.00 3.00 5.00
Group 3 5.00 4.00 4.00 5.00
Group 4 4.00 4.00 3.00 5.00
Group 5 4.00 4.00 3.00 2.00
Group 6 4.00 4.00 2.00 5.00
6 minutes 4.25±0.42 3.92±0.20 3.00±0.63 4.50±1.22

Mean ± SD
Group 1 4.50 3.50 3.00 5.00
Group 2 4.00 4.00 3.00 5.00
Group 3 5.00 4.00 4.00 5.00
Group 4 4.00 4.00 3.00 5.00
Group 5 4.00 3.00 2.00 5.00
Group 6 4.00 4.00 2.00 5.00
8 minutes 4.25±0.42 3.75±0.42 2.83±0.75 5.00±0.00

Mean ± SD
Group 1 4.50 3.50 3.00 5.00
Group 2 4.00 4.00 3.00 5.00
Group 3 4.00 4.00 4.00 5.00
Group 4 4.00 4.00 3.00 5.00
Group 5 4.00 3.00 2.00 5.00
Group 6 4.00 4.00 2.00 5.00
10 minutes 4.08±0.20 3.75±0.42 2.83±0.75 5.00±0.00

Mean ± SD
OPTIMAL TEMP. Optimal

Temperature
TIME TO 100% 8 minutes

HYDROLYSIS AT

OPTIMAL TEMP.

Graph 2: This graph shows that the optimal temperature of fungal amylase is at 55 C, and

it was reached at the 8 minutes interval.

 Fungal Amylase at 8 min
6

5
Color Change

0
0 25 55 85

Temperature (°C)

The table above showed the results for the experiment regarding fungal amylase. As the

reader can observe, 0 minutes is the control group because it remains constant (5), without any

color’s change from black to yellow when iodine was added, due to the fact that enzyme

amylases were not added to this group.

On the other hand, the group with 85 C temperature did not suffer any color change either

because the temperature was above to the optimal point for this enzyme which apparently is 55C.

Thus, the high temperature produced a denaturation of the enzyme. This caused a change of

shape of the enzyme ending in a change in its function too, producing no effect when it reacted

with the starch.

In addition, when the solution with starch and amylases was exposed to 0 C temperatures,

the color changed from black to dark brown when iodine was added. The reason was that the

temperature was too low and it caused a reduction in the interaction between subtracts and the

active site.
 However, when the solution test was exposed to 25C, it had a color change from black to

light brown due to fact that the temperature was not the optimal one, but it produced more

interaction between substrate and active side than the solution exposed to 0 C. Furthermore, it

did not affect the shape of the enzyme as it occurred when the substance was exposed to a

temperature of 85C.

Finally, the optimal point of enzymatic activity was reached at 55 0C after 8 minutes of

exposure to the solution of starch and amylase to that temperature. Therefore, this was the spot

plate that suffered the biggest change in color going from a black or dark blue (indicator of

starch’s presence) to a light yellow (indicator of no starch’s presence) because the enzyme broke

down the starch bonds.

Discussion:

There is correlation between the temperature´s change and the enzymatic activity because

color´s changes occurred in the reaction. The optimal point of enzymatic activity in fungus

amylases was reached at 55 C and the time of 100% hydrolysis at optimal temperature was

reached at 8 minutes of exposure of the solution of starch and amylase to 55 C (see table 5 and

figure 2). This was the spot plate that suffered the biggest change in color going from a black or

dark blue (indicator of starch’s presence) to a light yellow (indicator of no starch’s presence)

because the enzymes broke down the starch bonds.

On the other hand, the optimal point of enzymatic activity in bacterial amylases was

reached at 55 C and the time of 100% hydrolysis at optimal temperature was reached at 10

minutes of exposing the solution of starch and amylase to that temperature (see table 4 and

figure 1).. This was the spot plate that suffered the biggest change in color going from a black or

dark blue (indicator of starch’s presence) to a light yellow (indicator of not starch’s presence)

because the enzyme broke down the starch bonds.

However, fungus amylase is more sensible to the temperature than bacterial amylases, and

there optimum temperature should be 25C not 55C. As the reader could observe the 100 % of

hydrolysis was reached 2 minutes before in the reaction that contained fungus amylases than in

the one with bacterial amylases (see tables 4 and 5). The experiment´s results could be affected

by human errors such as: not well-mixed reactants after the enzyme was added, or the use of a

wrong temperature, or the wrong pipette. Furthermore, the room´s ambient temperature could be

improperly set, or the synchronization between the groups could also affect the results.

Therefore, some future recommendations should be to pay more attention during the experiment
to avoid not well-mixed reactants, use of the wrong pipette, or synchronization issues, and

measure all the temperatures before use, to be sure that they are the right ones.

Even though the experiment was not perfect, the data obtained in this experiment is

important because showed that enzymatic activity was affected by temperature and enzymes

have an optimum temperature point. When the temperature of the reaction exceeds this optimum

point causes a denaturation of the enzymes and a change in their shape what implies a change in

their function so the enzymes will not be able of work properly (Urry, Cain, Minorsky

Wasserman and Pearson Reece, 2016). The readers can observe this in the 85 C temperature

where the solution remains black in both source of enzymes, because the temperature was too

high and produced a denaturation of the enzymes (see figure 1 and 2).

Moreover, the data obtained supports that if the temperature of the reaction is too low the

substrates are not able of binding to the active site because the active site was not flexible

enough to fit with the shape of the substrate. In addition, the number of interaction between

substrate and active site is going to decrease (Urry, Cain, Minorsky Wasserman and Pearson

Reece, 2016). For example at 0 C temperatures, the reactions remained dark brown because the

enzymes did not work properly (see figure 1 and 2).

However, when the solution test was exposed to 25C, it had a color change from black to

light brown because the temperature was not the optimal one, but produced more interaction

between substrate and active side than the solution exposed to 0 C. Furthermore, it did not affect

the shape of the enzyme as opposed when the substance was exposed to 85C.

Therefore, during the process of breaking down starch using amylases if the temperature

increase or decrease drastically below or above of the optimal point of the specific enzyme

(amylase). The enzymatic activity is going to decrease or stop because the temperature affects
the enzymatic activity producing a denaturation of the protein that composes the enzyme. if is

too hot or if it is too cold is not going to be enough interaction between the substrate and the

active site for them to work.

In conclusion, enzymes are being used in the modern world in different areas such as

medicine, brewing processes, and baking. Enzymes are also present in signal transduction to

generate muscle contraction and organisms use them in metabolic process to obtain energy

Alberte J., Pitzer T., Calero K. (2012). Is important to know how enzymes react to temperature´s

changes and which are their optimum points of temperature in order for them to work properly.

For example, a doctor could recommend keeping some medicines that contain enzymes in a

room temperature instead of in a cool place or vice versa in order for the medicine to work

properly, and save lives.

 Literature Cited Section:

Alberte J., Pitzer T., Calero K., 2012, General Biology Lab Manual / Second Edition.

Florida International University: The McGraw Hill Companies: pp 49-51.

Garcia-Viloca M., Gao J., Karplus M. Truhlar D. G., 2004, How Enzymes Work: Analysis

by Modern Rate Theory and Computer stimulations. Science 303:pp. 186-195.

Harisha S., 2006, Introduction to Practical Biotechnology. India: Laxmi Publications.

Raven P., Johnson G. B., Mason K. A., Losos J. B., Singer S. S., 2008, Biology 8th

edition. New York: The McGraw Hill Companies.

Ringe D., Petsko G. A., 2008, How Enzymes Work. Science 320: pp. 1428.

Urry, Cain, Minorsky Wasserman and Pearson Reece, 2016, Campbell Biology In Focus.

Whitehurst R. J., Van Oort M., 2009, Enzymes in Food Technology: Wiley-Blackwell; 2

nd edition.