Enzymatic Activity Of

α-Amylase At Different
Temperatures
Auna Allen (Introduction)
Lyndsi Wilgenbusch (Methods)
Janice Hines (Results)
Mihajlo Gomez (Conclusion)
 Introduction
Determining the effect of temperature on the enzyme alpha-amylase.

Hypothesis: By Putting Amylase on Ice the temperature will stop the reaction;
causing the absorbance to lower than if it were in standard room temp.

What is an Enzyme? Enzymes are catalysts, usually proteins.

A Catalyst is a substance that speed up a chemical reaction without being

changed..

Enzymes bind to a substrate (the reactant molecule the enzyme works on) this
binding allows the reaction to occur more efficiently by lowering the activation
energy of the reaction.

The area where the Enzyme and the Substrate bind is called the active site.
(OpenStax College)
Enzymes bind to a substrate this
binding allows the reaction to occur
more efficiently by lowering the
activation energy of the reaction.
The area where the Enzyme and
the Substrate bind is called the
active site.At the Active Site the
Enzyme might change its shape to
fit with the substrate which forms
the enzyme/substrate complex

(OpenStax College)
Optimal pH and
Temp affect the
“lock and key”
concept of the
enzyme and
substrate
Without the correct pH and temp. The
enzyme could change shape causing the
substrate not to fit correctly, denaturing
the reaction.
Glycogen + Amylase = Glucose
(Substate) + (Enzyme) = Reaction
*Occurs if at optimal pH and temp.

(Ophardt)
Optimal Conditions of Enzymes
Enzymes work best within specific temperature and pH ranges, and optimal
conditions can cause the enzyme to lose its ability to bind to the substrate
because its change in shape.

Temperature: Raising the temperature generally speeds up the reaction.

Lowering the temperature slows down the reaction. While extreme temperatures
can cause the enzyme to lose its shape denaturing (stoping) the enzyme from
working correctly.

(OpenStax College)
 Introduction Continued
Based on our knowledge on enzymes we can predict that by
putting the enzyme on ice it will denature Amylase. This causes the
enzyme to change its shape incorrectly which the substrate would
then not “fit” into the enzyme correctly. We can make this prediction
because of the concept “lock and key”.
Results
- T Value- -4.988
- P Value- .000316
- Degrees of freedom- 12
- The p value is less than .05 so there is a significant difference between
temperature and absorbance.
- This means we reject the null hypothesis.
- Looking at figure 1, it is clear there is a significant difference in absorbance
between room temperature and ice.
- Figure 1 also shows us that at room temperature the absorbance steadily
increased while on ice there was hardly any increase.
Figure 1

Temperature and
absorbance did have
a significant difference
because .000316
<.05
 Methods
Steps:
1. Gathered Amalayes from our group members saliva.
2. Added .5ml of glycogen into 12 tubes and in the 13th
tube we added distilled water.

3. Added .5 ml of saliva solution

 Methods Continued...
4. 6 tubes were placed into ice baths and 6 were left in
room temp

5. After 8 minutes DNSA was added to stop the reaction

6. Boiled tubes for 5 min, then added 18 mL of distilled

water.
 Methods Continued…
● Independent Variable: Temperature
● Dependent Variable: Absorbance
● Standardized Variables: Timing of DNSA
● Modified the TEMPERATURE to test hypothesis
● Concentration of Glycogen: .5ml
Discussion/Conclusion

- There is a clear difference in absorbance between the enzymes put in

ice and the enzymes left at room temperature.
- The P value is lower than .05 meaning there is a significant difference
in the absorbance of glucose at room temperature and ice.
- Our groups hypothesis was correct because the test tubes put in ice
had less absorbance compared to the test tubes left at room
temperature.
Discussion/Conclusion Continued...
Other questions that arose.

- Would heating up the test tubes cause a difference in absorbance?

- If more glycogen is added to the solution, would there be a difference
in absorbance
- What is the ideal temperature to give the enzymes optimal
absorbance?
Discussion/ Conclusion Continued...
- In a study done by Laurent Bleuze they studied the absorbance of
enzymes in hemp under controlled air temperature, humidity, and with
simulated rain.
- They found that there was a difference in enzyme absorbance
depending on temperature, humidity, and rain
- This relates to our study because temperature had an effect on
enzyme absorbance in both studies.
 References
Bleuze, L., Lashermes, G., Alavoine, G., Recous, S., & Chabbert, B. (2018). Tracking the dynamics of hemp dew retting
under controlled environmental conditions. Industrial Crops & Products, 123, 55. Retrieved from
http://link.galegroup.com.ezproxy.loras.edu/apps/doc/A553489173/AONE?u=lorascoll&sid=AONE&xid=1cef28a4

OpenStax College. 2019 "Potential, kinetic, free, and activation energy"by Biology, CC BY
3.0._https://www.khanacademy.org/science/biology/energy-and-enzymes/introduction-to-enzymes/a/enzymes-and-the-activ
e-site Accessed February, 26 2019

Ophardt, Charles. 2003. Virtual Chem Book, Elhmhurst college http://chemistry.elmhurst.edu/vchembook/571lockkey.html

Acessed Febuary 26, 2019

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Enzymatic Activity of
α-Amylase At Different
Temperatures