Amylase Hypothesis

Rubric Enzyme Report (40 points)
1. Introduction: (4 pts)
a. What are enzymes and what do they do?
b. What are the optimal conditions for an enzyme?
c. What factors affect enzyme activity and how?
d. How is the structure of an enzyme related to its function?
e. What is amylase? What reaction does it catalyze?
f. Research the amylase enzyme you used (organism) and the optimal conditions of
temperature, pH, or salt, it requires.
g. Explain why DNS is used and why it is important for determining enzyme activity.
h. Cite and reference your sources using APA style.
2. Experimental Design: (5 pts)
a. State your question, hypothesis (ensure that it is testable and falsifiable), and prediction.
b. Identify the independent variable.
c. Identify the dependent variable.
d. Identify the experimental group
e. Identify the control group.
3. Materials/Methods: (3 pts)
List all of the equipment and materials you will need for your experiment. BRIEFLY
describe your experimental procedure for setting up the maltose standard curve (Part II) and
for testing your hypothesis on amylase (Part III).
4. Data Analysis/Results:
a. List all the formulas you used for your calculations and a sample calculation in each case
(3 pts)
b. Prepare the following tables:
• Table 1 with maltose standard curve data. (3 pts)
• Table 2 with your experimental setup. Explain how your control tubes will be set up.
(2 pts)
• Table 3 with absorbance measurements of experimental tubes, calculated maltose
concentration, calculated amylase activity (mg/ml maltose/min) (4 pts)
c. Prepare the following graphs:
• Graph 1 of maltose standard curve: show line of best fit and equation of line if done
on excel. If done manually, insert an image of your graph indicating the line of best
fit. (3 pts)
• Graph 2 of enzyme activity versus independent variable (4 pts)
d. Summary of results: summarize your results based on your graphs (3 pts)
5. Conclusion/Discussion: (4 pts)
Based on your results, is your hypothesis supported? Restate your hypothesis and explain!
Discuss any other findings and explain them. Discuss errors, and reasons for data variability.
6. References: (2 pts)
List the sources you used at the end of your report, and include in-text citations in your report
(APA citation style)

ANSWER

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Amylase Hypothesis

  1. Introduction: (4 pts)
    1. What are enzymes and what do they do?

An enzyme is a type of protein found in a cell responsible for speeding up the rate of chemical reactions to support life.

  1. What are the optimal conditions for an enzyme?

The optimum conditions for an enzyme is a warm temperature of around 35 to 400C and an absolute pH.

  1. What factors affect enzyme activity and how?
  1. Temperature – Absolute temperature enable optimum enzyme activity where if too cold, enzymes will do nothing and if too hot, considering they are proteins, they get denatured.
  2. pH – Changing the pH will affect the charges on the amino acid molecules in the enzymes.
  • Enzyme concentration – The more the enzymes the faster the reaction until saturation.
  1. Substrate Concentration – Increase in Substrate Concentration increases rate of reaction until all enzymes are bound then there will be no effect.
  2. Presences of Inhibitors or Activators.
    1. How is the structure of an enzyme related to its function?

An enzyme binds with the appropriate substrate only when the correct alignment and orientation to connect the molecules.

  1. What is amylase? What reaction does it catalyze?

Amylase is a special enzyme that is involved in the digestion reactions of food.

  1. Research the amylase enzyme you used (organism) and the optimal conditions of temperature, pH, or salt, it requires.

Maximal activity of the enzyme amylase is at 7.0 pH, optimal temperature of 370C and 10% salinity.

  1. Explain why DNS is used and why it is important for determining enzyme activity.

Dinitro Salicylate is important in detecting reducing sugars under the action of hydrolase enzymes carbohydrates.

  1. Cite and reference your sources using APA style.

Kolusheva, T., & Marinova, A. (2007). A study of the optimal conditions for starch hydrolysis through thermostable α-amylase. Journal of the University of Chemical Technology and Metallurgy42(1), 93-96.

 

  1. Experimental Design: (5 pts)
    1. State your question, hypothesis (ensure that it is testable and falsifiable), and prediction.

Presence of an enzyme speeds up a biochemical conversion of a substance.

enzyme

Substrate         →                   Product

In this experiment, amylase is the enzyme while starch is the substrate.

  1. Identify the independent variable.

Presence or absence of amylase

  1. Identify the dependent variable.

Presence or absence of glucose

  1. Identify the experimental group

 

 

  1. Identify the control group.

Quantities of the solutions used, the time taken before testing and the temperature of the water

  1. Materials/Methods: (3 pts)

List all of the equipment and materials you will need for your experiment. BRIEFLY describe your experimental procedure for setting up the maltose standard curve (Part II) and for testing your hypothesis on amylase (Part III).

Materials.

  1. Amylase Solution
  2. Boiled Amylase solution in boiling tubes
  • Starch Solution
  1. Starch Powder
  2. Glucose Powder

Reagents and Apparatus

  1. Iodine Solution
  2. Benedict’s Solution in Dropper Bottles
  • 8 clean test tubes
  1. 1 glass rod
  2. I recessed tile
  3. Paper towel
  • Spirit Marker
  • Safety Glasses
  1. Plastic Pot

 

  1. Data Analysis/Results:
    1. List all the formulas you used for your calculations and a sample calculation in each case (3 pts)
    2. Prepare the following tables:
      1. Table 1 with maltose standard curve data. (3 pts)
    S-Starch M-Maltose G-Glucose
Solubility   Cloudy Clear Clear
Colour with Iodine Blue-black Clear Orange Clear Orange
  Benedicts Blue Orange ppt Orange ppt

 

  1. Table 2 with your experimental setup. Explain how your control tubes will be set up.(2 pts)
Tube Contents Results
    Iodine Test Benedict’s Test
1 2.5 ml amylase solution and 2.5 ml start solution Browny-orange Bright orange ppt
2 2.5 ml boiled amylase solution and 2.5 ml start solution Blue-black Clear blue
3 2.5 ml amylase solution Orange Clear blue
4 2.5 ml boiled amylase solution Orange Clear blue
5 2.5 ml starch solution Blue-black Clear blue

 

  • Table 3 with absorbance measurements of experimental tubes, calculated maltose concentration, calculated amylase activity (mg/ml maltose/min) (4 pts)

 

  1. Prepare the following graphs:
    1. Graph 1 of maltose standard curve: show line of best fit and equation of line if done on excel. If done manually, insert an image of your graph indicating the line of best fit. (3 pts)

 

  1. Graph 2 of enzyme activity versus independent variable (4 pts)

 

 

  1. Summary of results: summarize your results based on your graphs (3 pts)

 

  1. Conclusion/Discussion: (4 pts)

Based on your results, is your hypothesis supported? Restate your hypothesis and explain!

The hypothesis that presence of an enzyme speeds up a biochemical conversion of a substance is true

Discuss any other findings and explain them. Discuss errors, and reasons for data variability.

 

 

  1. References: (2 pts)

List the sources you used at the end of your report, and include in-text citations in your report (APA citation style)

Kolusheva, T., & Marinova, A. (2007). A study of the optimal conditions for starch hydrolysis through thermostable α-amylase. Journal of the University of Chemical Technology and Metallurgy42(1), 93-96.

Deb, P., Talukdar, S. A., Mohsina, K., Sarker, P. K., & Sayem, S. A. (2013). Production and partial characterization of extracellular amylase enzyme from Bacillus amyloliquefaciens P-001. SpringerPlus2(1), 154.

 

 

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