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Describe three different factors that can impact enzyme activity.
Temperature
pH
presence or absence of cofactors
Describe what happens to the physical structure of enzymes at suboptimal conditions using proper scientific terminology
Denaturation occurs due to the enzyme unfolding and losing its shape
Extremely high temps can disrupt the bonds and interactions between amino acids in a protein
Extremely low temps can make the enzyme more properly folded but molecules move more slowly so reaction rates are slowed
Define a colorimetric test and give an example
Any test in which a color change is used to indicate the presence of a chemical or molecule
Describe the color or Lugol’s iodine in the presence of starch
Is a blue-black color in the presence of starch
Define type of macromolecule for each of the following:
a. Starch
b. Amylase
c. Maltose
Starch is a polysaccharide, which is a type of carbohydrate. It is composed of long chains of glucose molecules linked together by glycosidic bonds and serves as a storage form of energy in plants.
Amylase is a protein, which is a type of macromolecule made up of amino acids. It functions as an enzyme that catalyzes the breakdown of starch into smaller sugar molecules, like maltose.
Maltose is a disaccharide, which is a type of carbohydrate. It consists of two glucose molecules linked by a glycosidic bond and is a product of starch digestion.
Explain how you can test amylase activity to determine optimal temp and pH conditions for the enzyme
For optimal temp, you will combine amylase w/ a starch solution in a microfuge tube then you will test for starch digestion by adding the amylase/starch mixture to Lugol’s iodine and note the color change
For pH conditions, you will prepare starch and iodine
Identify what color the following reactions should turn:
1% starch + functional amylase + Lugol’s iodine
1% starch + nonfunctional amylase + Lugol’s iodine
Pale yellow
Blue-black
For E1 and E3, explain what a black/brown tells you about amylase activity. Explain what a yellow color at the end of the exercise tells you about amylase activity
black/brown = amylase is not breaking the starch down effectively (low activity)
yellow = amylase activity is active
Explain the relationship between H+ concentration and the pH scale.
The lower the # on the pH scale, the higher the concentration of hydrogen ions in the solution
Explain what happens to the hydrogen ion concentration of a solution when acids or bases are added
When an acid is added, the concentration of hydrogen atoms increases. The pH also decreases
When a base is added, the ph increases and the concentration is decreasing.
Demonstrate the ability to determine the pH of a solution using a pH meter
Get pH meter and measure the solution
Differentiate between acids, bases, and neutral solutions
Acids are below 7
Bases are above 7
Neutral is 7
Define buffer and buffering capacity and give a biological example.
Buffering capacity is measured by how much a solution resists change in its pH when acid or base is added. It requires a weak base and acid.
A solution has a high buffering capacity when it resists change in pH when an acid or base is added
A solution has a low buffering capacity when the pH changes fairly quickly when an acid or base is added
An example is carbonic acid
Explain why buffers are important in biology
It’s important because it helps maintain pH
Describe how the carbonic acid buffer system works.
Carbonic acid acts as a weak acid, able to donate protons when a base is added.
When acid is added, the solution is able to absorb the protons using the conjugate base, the bicarbonate ion, reforming carbonic acid.
The effect on the pH of the solution is minimized by the presence of the weak acid and conjugate base pair
if pH goes up, the rate of the forward reaction will increase to release H+ into the solution
if pH goes down, the rate of the reverse reaction will increase to absorb H+ from the solution