Amylase Investigation
* Hypothesis
I predict that as the temperature increases, the speed of the reaction
will increase. When a particular temperature is reached I believe the
rate of reaction will dramatically decrease. I believe this because
most chemical reaction happens faster when the temperature is higher.
At higher temperatures molecules mover around faster, which makes it
easier for them to react together. Usually, a rise of 10OC will double
the rate of reaction. This is true for enzymes up to about 40OC.
However at 40OC the enzyme begins to be damaged, so the reaction slows
down. By 60OC the enzyme is completely denatured. I predict that the
same will happen the further away the pH is from pH 7. I believe this
because the optimum temperature for most enzymes is about pH 7
therefore the further away from pH 7 (either more alkaline or more
acidic) the less affective the enzyme.
* Pilot Experiment
To investigate the factors affecting the activity of the enzyme
Amylase, I will adapt a pilot experiment, which investigates if the
enzyme amylase breaks down starch. The pilot experiment was:
* Pour amylase solution into a test tube to a depth of 2cm.
* Half fill another test tube with a 4% starch solution.
* With a pipette place a drop of iodine into each dimple in a dimple
tray.
* With a glass rod lift a drop of the starch solution from the test
tube and mix it with the first drop of iodine in the first dimple in
the tray. A blue/black colour should develop; this will be used as the
control.
* Rinse the glass rod.
* Pour amylase solution into the test tube of starch and shake
quickly.
* Repeat steps 4 ...
... middle of paper ...
...ting the
experiment. However, they should have been left in the water bath for
about 30 minutes so that the amylase had been completely affected by
the temperature before the experiment was started.
I decided to conduct the experiment at 10 OC intervals instead of 5 OC
because there was not enough time. When the results were collected I
plotted them on a rough graph to find the optimum temperature and then
conduct the experiment at this temperature to ensure it was the
optimum temperature. I also conducted all three experiments for each
condition at the same time to save time.
Additional work, which could be carried out, is to repeat the
experiment using, a wider range of temperatures and pH levels, a range
of different starch solution concentrations or using different enzymes
such as protease with a protein.
3.) Divide your 30g of white substance into the 4 test tubes evenly. You should put 7.5g into each test tube along with the water.
The first test was to put 5 drops of the distillate into a test tube
I blanked it with 2 cm³ water, 1 cm³ amylase and 3 drops of iodine.
1. Using a dropper, add a drop of the Benedicts Reagent into a test tube.
Investigating Amylase Aim: The aim of the experiment is to investigate how effectively the enzyme amylase breaks down starch at different temperatures, and therefore to find the optimum temperature that amylase digests starch. Introduction: Thousands of chemical reactions take place in our cells and those reactions need to happen quickly in order to keep us active. These chemicals are called ENZYMES. Enzymes make reactions happen at a much faster rate. Enzymes come in two main types, breakers and builders.
2. With the pipette take two drop of red dye and drop it into the
To carry out the experiment starch-agar Petri dishes are needed. Gloves must be worn to prevent contamination of the Petri dishes and so not to hinder the results.
4. Pour about 300mL of tap water into the beaker. Set up a hot-water bath using a hot plate, retort stand, and thermometer clamp. Alternatively, use a Bunsen burner, retort stand, ring clamp, thermometer clamp, and wire gauze.
The hypothesis is supported by the experiment in that with increased starch concentration, the amylase activity increased each time and the salivary amylase functioned best at higher concentrations of starch. Also, for the most part, the reaction followed the general trend of how at lower concentrations, the increase in reaction rate is greater, while at higher concentrations, the increase in reaction rate is less. Despite some discrepancies in trend, specifically at the 40g/L concentration, figure 1 still displays how the amylase activity eventually reaches a plateau, as mentioned in the hypothesis. From the results it appears that the point of saturation for this reaction is at the concentration 50g/L as the amylase activity rates for 50g/L
ice-cold calcium chloride to each tube and place both tubes in a beaker filled with ice.
Clamp the 50ml burette to the retorted stand and add the sodium hydroxide solution (base) using a funnel.
Questions & Tables from Experiment Three 1. What is the optimal pH for salivary amylase activity and how does it compare with the pH at which the enzyme is normally found? • The optimum pH level for salivary amylase is approximately 6.8. Any change of pH below or above the optimum pH decreases the rate of enzyme action. The changes in pH can lead to changes in the ionization state of the substrate or the enzyme or both.
Remove the extra solvent on a steam bath under a hood while flushing the flask with N2 gas, leaving the crude extract. Weigh extract.
Weigh out two 0.100 g. samples of the product and put each into a test
2nd step heat the mixture: Make sure the agarose dissolves. Wait until it boils and when you are going to transfer the mixture, wear gloves to avoid getting burnt. Transfer the mixture into a removable gel tray.