Investigate the Effect of Temperature on Amylase Activity, Lab Report Example

Design of an Experiment to Investigate the Effect of Temperature on Amylase Activity

Introduction

Amylase is acknowledged as an enzyme which motivates the decomposition of starch into its components. The component of starch is sugar. Amylase is encountered in nearly every plant, microorganism and animal species. Substantial quantities of amylase are found in the saliva and pancreases of mammals and in the germination processes of spores (Landersberg 190; Zajoncová 245).

Research Question

The research question for this experiment has the objective of   answering the following inquiry: How does the saturation of the substrate influence the reaction rate of amylase?

Aim

The aim of this experiment is to establish a relationship between the amount of substrate and the amylase activity. The amylase activity will be assessed by the amount of gas released the relationship with regards to increasing the amount of substrate on the amylase reaction? What are the effects of decreased amounts of substrate on the amylase reaction? The research questions will be answered in the outcome of the laboratory experiment.

General Knowledge

Enzymes are well acknowledged for their qualities as biological catalysts. Nearly all of the reactions which occur in cells are administrated by enzymes. The enzymes are composed of intricate three dimensional proteins which are globular in aspect. Many of the enzymes are made out of proteins and supplementary molecules which are associated with them. Notwithstanding that the enzymes possess a larger dimension than the substrate molecule; a minute portion of the enzyme comes into contact with the substrate molecule. This area of contact is defined as the activation zone. Enzymes have the capacity of accelerating the metabolic responses of the cells. Enzymes can be said to be substrate specific due to the aspect that not every enzyme is compatible with the activation zone of a substrate. The enzyme reactions are bi directional and the enzyme can be applied various times to a substrate (Landersberg 190; Zajoncová 245).

Hypothesis

Independent variable

• Amount of substrate.

Dependent variable

• Amount of gas liberated by the amylase – hydrogen peroxide reaction.

Controlled variables

• Weight of the potato- 180 g ± 0.5 g.
• Quantity of Substrate.

The independent variable in the experiment will be the saturation of the substrate. The dependent variable will be the quantity of oxygen which is liberated in the reaction. The controlled variables will be the weight of the potato, thermal characteristics of the reaction and the amount of substrate which will be applied. The controlled variable of the potato’s weight will be controlled by the application of a scale which can assess a mass as great as 2 d. p. in order to guarantee that all of the potato fragments which will be applied in the experiment will have individual masses which are equivalent to 180 g with an uncertainty of ± 0.5 g. The thermal characteristics of the reaction will be administrated by the application of a thermometer in order to assess the temperature of the room. A graduated cylinder will be applied in order to distribute the 10 ml of H₂O₂ that is required in order to conduct each trial of the experiment (OMSI 1).

The perspective which will be applied is to insert the potato fragments into test tubes and observe the reaction. In the bubbling of the substrate in the test tube, gas is liberated into the upside down graduated cylinder which is placed in the rinsing bowl. A number of comparisons will be made in order to find out the effect of the saturation of the hydrogen peroxide on the amylase reaction from the potato fragments (OMSI 1).

Apparatus and Methods

1. Test tube which will be applied for boiling the potatoes.
2. Rubber tubing.
3. Rinsing bowl.
4. Water (distilled).
5. 10 ml graduated cylinder.
6. 20 ml graduated cylinder.
7. 60 ml of hydrogen peroxide solution with 1%, 2%, 3%, 4% %% and 6% concentrations.
8. Clamp for the test tube which is applied for ebullition.
9. Cork carver.

The following steps should be conducted in order to perform this experiment:

1. Thirty six potato cylindrical fragments will be carved by using the cork carver.
2. Each of the potato cylinders which have been carved will be evaluated in order to guarantee that the potato cylinders have a weight of 180g ± 0.5g. The file and the knife will be applied in order to reduce the edges of the potato cylinders.
3. A rinsing bowl will be half filled with water.
4. The graduated cylinder will be applied in order to sasses and distributed 10 ml of 6% hydrogen peroxide.
5. The 10ml of 6% hydrogen peroxide will be distributed to the test tube which is applied for ebullition.
6. The 10 ml graduated cylinder will be immersed into the mixing bowl, filled with water and turned upside down. The air bubbles should be evacuated from the upside down graduate.
7. The segment of rubber tubing will be inserted into the rubber bung. The bung will be inserted into the 10 ml graduate cylinder.
8. A potato cylinder which had been previously carved and weighed to a weight of 180 g ± 0.5 g will be inserted into the test tube which is used for ebullition. The bung will be inserted into the test tube which is applied for ebullition in order to ensure a tight fit.
9. The chronometer is activated the instant that the potato cylinder is inserted into the test tube which is applied for ebullition.
10. Subsequent to the passing of 120 seconds, the rubber tubing should be extracted from the graduated cylinder.
11. The quantity of water which has been displaced by the graduated cylinder will be assessed.
12. The experiment is to be repeated five additional times with the identical concentration of hydrogen peroxide solution.
13. The steps which are numbered from four to twelve will be repeated for the 5%, 45, 3%, 2% and 1% concentrations of H₂O₂
14. The data which will be collected from the experiment will be entered into the following table.

The response rate between the starch and the amylase was affected due to the aspect that it may require more time for the breakdown of the starch molecules in the event that there is more starch present in the solution. The needed time for the dissolving of the blackened color in the test tube may have been the reason of the high standard deviation (Landersberg, 190; Zajoncová 243). In the experiment, one of the requisites was to completely rinse and dry out the test tube which was applied for boiling the potatoes after each of the trials. This was required in order to ensure that the saturation is not influenced by the previous solutions and saturations in the test tube which was applied for boiling the potatoes. It was important to supervise the thermal characteristics of the experiment in order to guarantee that there had been no abrupt changes so as not to influence the outcome of the experiment.

The response is a heat releasing reaction; this implies that there was a bubbling, heat releasing response. The heat energy which had been released had the potential of influencing the outcome. In the event of all of the controlled variables being properly managed, the assumption is that the quantity of heat energy which is released in each of the trials is equivalent (Landersberg, 190; Zajoncová 243). The study must be conducted a number of times in order to manage the outcomes. Consequently, it would better to conduct each trial a number of times in order to compare if the outcomes were reliable and if any erroneous outcomes may have been derived.

In addition, there are a minimal number of five outcomes which must be gathered in order to find out the standard deviation. The aspect of standard deviation is significant in this study.  The standard deviation lets the researcher to reach the conclusion of the experiment being reliable or not.

In trial 1, the amount of gas is collected will be entered on the table. The trials will be conducted five times for each of the saturations of H₂O₂. In trial number two the amount of substrate will be increased to a 2% concentration. This trials will be conducted five times  This amount of gas which will be collected in the upside down graduated cylinder which was positioned in the rinsing bowl  will be measured and  entered on the data table above.. In trail number three the concentration of the substrate will be increased to 3%. The amount of gas which will be collected in the upside down graduated cylinder which is in the rinsing bowl will be measured and entered on the data table above. This process will continue through the sixth trial until a substrate concentration of 6% is reached. Consequently, the amount of gas which will be collected in the graduated upside down cylinder which was placed in the rinsing bowl filled with water was assessed at an average of the five trials for each concentration. The results of the experiment will be used to confirm the research questions.

Works Cited

Landersberg, Joseph. Basic laboratory experiments for general, organic and biochemistry. Belmont, CA; Cengage Learning, 2011. Print.

OMSI. “Experiment potato power Unit 3 Biochemistry experiencing chemistry. OMSI, 1997. Web 2 March 2 2014 <http://www.omsi.edu/sites/all/FTP/files/chemistry/emeritus/MM-U3.0337>

Zajoncová, Ludmila. Milan Jílek, Vertonika Beranová, & Pavel Pe?. A biosensor for       the determination amylase activity. Biosensors and Bioelectronics, 20.2(2004):    240- 245. 2 March 2014 http://www.sciencedirect.com/article/pii/S0956566304000223