Bio 1 M1L2 Biological Macromolecules and Enzymes Lab Report 051520 (1)

PLAN UP TO 3 HOURS FOR THIS LAB – if you want to split it up it would be convenient to do Activities 1-3 one day (approximately 1 hour) and Activities 4and 5 on a second day (approximately 1 hour each).

Read the Background provided on the lab instructions, information from your textbook (if applicable – if you are taking this course independently of a corresponding lecture course I recommend finding a good free, online biology textbook as reference), and any information provided in the READ area of the module to help you answer these pre-lab questions:

**List at least 2 monomers for each of the following macromolecules a. Polysaccharide (like starch/amylose) – glucose, fructose b. Triglyceride – fatty acids, glycerol
**List 1 monomer for each of the following macromolecules a. Protein – amino acid b. Nucleic acids - nucleotides
**Which of the four macromolecule groups do enzymes belong to? a. Proteins
**Name two environmental conditions that can affect enzyme action. a. Temperature and pH
**Research and define the term “denaturation” in your own words. Include the following in your explanation (along with and an appropriate citation and reference. Denaturation occurs when a protein’s original shape is changed due to factors such as pH and temperature. Denaturation causes the protein to unfold and deviate from its normal/original shape. a. Which class of macromolecules is sensitive to denaturation i. Proteins are much more sensitive to denaturation than any other macromolecule due to their unique 3D shape. b. What conditions can cause denaturation? i. Heat, pH changes, and heavy metal ions can all contribute to denaturation. c. What happens to the function of these molecules when they are denatured? Once a protein is denatured, it loses its ability to function as it properly should. It can no longer complete the functions it is assigned to.
**Research what a “positive control” and a “negative control” are and the difference between them. Briefly explain them in your own words (2-3 sentences) and provide an appropriate citation and reference.

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c. MAKE SURE YOU SAVE any leftover IKI – we’ll use it next week. 11. Insert your pictures below the data tables.

Data Table 1 (expanded)

Test Tube ID Observations

Test tube 1 Water+iodine potassium iodide

Hypothesis: NO REACTION (negative control)

Result: No reaction (yellow) Test tube 2

Water+potato starch+iodine potassium iodide

Hypothesis: REACTION (positive control)

Result (color): Positive (black ish)

Test tube 3 Water+albumin solution+iodine potassium iodide

Hypothesis: No reaction

Result (color): Negative (yellow)

Common Item #1: (milk)

Result: Yellow Common Item #2: (Olive oil)

Hypothesis: No reaction

Result: Brownish Common Item #3: (Lime juice)

Hypothesis: No reaction

Result: Yellowish brown

**How did you use the positive and negative controls to interpret your results from the experimental tubes (albumin and common items 1-3)? a. I was able to interpret the results by having a guide as to what the positive and negative controls looked like. If my result was browner than yellow I would consider that a positive result as the iodine reacted with the test subject.
**Did any of your results surprise you? If they did please describe and explain if possible. I was not expecting milk to have starch in it, I normally consider it more of a dairy product but now when I think about it, it makes sense as milk is also used to make things that have starch in them. I also was not expecting lime juice to contain starch, but it seems there is a little bit of starch in there. Adapted for Empire State College from

Photo 1

Insert the photo from Activity 1, step 5. The following should be indicated in this photo (we should either see the tubes clearly labeled or you should add labels or writing on your image so we can distinguish between them):  Test Tube 1 – water  Test Tube 2 – starch  Test Tube 3 – albumin

Photo 2

Insert the photo from Activity 1, step 8 (repeated step 5) for your additional food samples. Clearly indicate which food is in which test tube.  Common Item #1 – Milk  Common Item #2 – Olive oil  Common Item #3 – Lime juice

Adapted for Empire State College from

b. TAKE A PICTURE of step 5 and step 8 16. Insert your pictures below the data tables.

Data Table 2 (expanded)

Test Tube ID Observations

Test tube 1 Water + Biuret reagent

Hypothesis: no reaction

Result (color): light blue Test tube 2 Water+albumin solution+Biuret reagent

Result (color): Purple Test tube 3 Water+potato starch solution+Biuret reagent

Hypothesis: No reaction

Result: Light blue Common Item #1: (Milk)

Hypothesis: No reaction

Result: reaction - purple Common Item #2: (Olive oil)

Result: Mild reaction - dark blue

Common Item #3: (Lime juice)

Hypothesis: no reaction

Result: No reaction.. hard to tell.. light blue ish

**Did any of your results surprise you? If they did please describe and explain if possible.

Photo 3

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Photo 4

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Data Table 3

Test Tube ID Observation

Hypothesis: No reaction

Result (opaque (-) or translucent (+): opaque

Vegetable or olive oil

Result (opaque (-) or translucent (+): Positive Vinegar Hypothesis: Positive Result: Positive Salad dressing Hypothesis: Positive Result: Positive

Result: Positive Lime juice Hypothesis: Negative

Adapted for Empire State College from

ii. **Hypothesis 2: If an acid and an enzyme is present but no heat is present, no digestion will occur and the protein will still be present after incubation. 1. White tubes should we compare to make conclusions about this hypothesis? 1b and 2b b. **Do you predict that tubes 1a & 2a will test positive or negative for protein in step 4? Do you predict that tubes 1b & 2b will test positive or negative for protein in step 10? i. **PUT YOUR PREDICTIONS IN TABLE 4 – INSERT AN IMAGE OF YOUR TABLE BEFORE THE ACTIVITY WITH THESE PREDICTIONS HERE:

Photo 6

Insert the photo of your predictions here:

Use the instructions provided to complete Activity 4 and record your results in the data table below. c. TAKE A PICTURE of ALL FOUR test tubes at steps 6 & 11
Insert your pictures below the data tables.

Data Table 4

Test Tube ID Observation

Test tube 1a Albumin+pepsin+HCl+Biure t reagent before incubation

Result: positive Test tube 2a Albumin+pepsin+water+Biu ret reagent before incubation

Result: positive Test tube 1b Albumin+pepsin+HCl+Biure t reagent after incubation

Result: negative Test tube 2b Albumin+pepsin+water+Biu ret reagent after incubation

Photo 7

Insert the photo from Activity 4, step 6, The following should be indicated in this photo:

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 Test Tube 1a  Test Tube 1b  Test Tube 2a  Test Tube 2b

Photo 8

Insert the photo from Activity 4, step 11, the following should be indicated in this photo:  Test Tube 1a  Test Tube 1b  Test Tube 2a  Test Tube 2b

Adapted for Empire State College from

experiment to rule out the effect of time of your outcome (describe in 3-5 sentences)?

I do think the outcome of the experiment is influenced because one set is tested immediately. I am interested to see if the heat of all tubes at the same time would change the digestion process. There was no acid in the two tubes so I don’t know if the heat would help them or not in the digestion process.

Activity 5

BEFORE YOU BEGIN: read through the activity, collect your materials, and respond to the following: a. ***In this activity, what is the substrate and what/where is the enzyme? The enzyme is amylase which is in our saliva. The substrate is starch. b. **Write out a hypothesis that this experiment will test. If the cracker is chewed, there will be a positive result because the enzyme amylase will break down the carbs and initiate digestion.

Data Table 5

Test Tube ID Observations

Cup 1 Cracker + DI water + IKI

When the IKI was put in it splattered out on the cracker. It is a lot less consistent. It is more of a purpleish color Cup 2 Cracker + saliva + IKI

When the IKI was put it in melted into the cracker. It is more of a dark brown/purpleish color

Photo 9

Insert the photo from Activity 5, step 9. The following should be indicated in this photo: I couldn’t use the foam cups because mine were broken.

Adapted for Empire State College from

***What do you think would happen if the saliva treatment of the starch (cracker) was conducted under very cold conditions? Very hot conditions? I think if the experiment was conducted under cold conditions it would take the cracker much longer to dissolve and digest. I believe if the experiment was conducted under hot conditions, digestion and dissolution would occur much more rapidly as heat speeds up digestion.

Consider Activities 4 & 5 and the hypothesis below. Develop your responses to the following questions into a 1-2 paragraph DISCUSSION as in a formal scientific paper (INCLUDE AT LEAST 2 PROPERLY CITED & REFERENCED SOURCES – you may use sources that were used previously in this lab).

Hypothesis: Enyzmes require specific temperature conditions in order to effectively carry out their reactions with substrates.

Adapted for Empire State College from

**Why is it important to understand the relationship between temperature and enzyme action (relevance to humans and/or human systems)? Understanding the relationship between temperature and enzyme action becomes important when discussing things such as digestion. It is known that an enzyme binds to a substrate but the process after and the manipulating factors sometimes are unknown. It is important to note that, “the temperature for denaturing is specific for each enzyme” (Effect of Temperature on Enzymatic Reaction, 2022). Denaturing happens for enzymes around freezing temperature and there is no getting the enzyme back after it reaches this point. The article Enzyme Function Dependent on Temperature tells us that “When the temperature is too hot or too cold, or unsteady, the enzymes will spend less time in their optimal shape which simply translates into having a less than optimal metabolism” (Enzyme Function Dependent on Temperature, nd).). So, if an enzyme is not equated with optimum temperature, it will not function correctly. One interesting example of this arises when looking at patients that present with fevers. It is stated that, “If the temperature goes too high, the patient’s brain enzymes can denature, which can cause delirium and can even be life threatening” (Enzyme Function Dependent on Temperature, nd). So we don’t only see the importance of the enzyme temperature relationship in the digestion process but also in many other parts of our body.
**Did your results in Activity 4 & 5 match your predictions? If they did not, provide a possible explanation for the results you observed (interpretation of your results). Yes, for the most part my predictions were supported by my experiment. For activity 4, it makes sense that the tube with a protein, an enzyme, an acid and heat was the tube that showed digestion occured. This makes sense because it contained all of the required materials for digestion. We saw that without one of the parts, such as heat, the enzyme did not aid in digestion and no digestion occurred. For activity 5, after adding the iodine, it seems that both crackers went through the process of digestion. Now, I am not sure if this is correct because the cracker without chewing still had a little bit of black once the iodine was put in. I think this could mean that some digestion occurred so this was a little surprising to me because no heat was applied so I wouldn’t think the digestion would occur because no enzyme was present. There could have been slight errors in the experimental procedure.

Adapted for Empire State College from