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Find out how temperature affects the enzymic action of urease on urea to produce Carbon dioxide and ammonia.

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Introduction

My investigation I have carried out a research on the type of enzyme that I have chosen to use, urease. Urease is an enzyme that catalyses the hydrolysis of urea into ammonia and carbon dioxide. The reaction occurs as follows: It is a globular protein found in bacteria and several higher plants as found by James Sumner in 1926. Its molecular weight is 480kDa with optimum pH of 7.4 and its enzymatic specificity is urea and hydroxyurea. Urea is the chief solid component of mammalian urine; synthesized from ammonia and carbon dioxide and used as fertilizer, in animal feed and in plastics. Preliminary Work I have carried some preliminary experiment to help me decide on how to measure measure the effect of temperature on the activity of urease. In the first experiment, I used the universal indicator to find out if temperature has on effect on the pH of the ammonium produced by the reaction. In the second experiment I used the litmus paper but it was still the same effect. I found out that it does have an effect but not a lot and it was hard to measure as the colour was always green although in different shades. In the third one I used the electronic pH indicator but the pH numbers hardly changed. I decided on using the electronic pH indicator but the pH numbers hardly changed. ...read more.

Middle

>To collect the gas in. >To filter urease >To thoroughly filter urease >For accuracy when making measurements of the urea and urease. >To time the experiments Apparatus, Chemicals and their Function Chemical Function >Urea >Urease >Distilled water >To react with urease to produce ammonia and carbon dioxide, this will enable me to how fast it is when reacted together >To catalyse the hydrolysis of urea into ammonia and CO2. >To make the urea and urease 5% concentrations Hazards If swallowed: Wash out mouth and give a glass or two of water and seek medical attention. If solid gets into eyes: Flood the eye with gently running water and wash thoroughly with soap and cold water. If spilt in lab: Scoop up as much as possible. Wash area thoroughly with detergent and water. If spilt on skin or clothes: Remove contaminated clothing. Flood area with water and wash thoroughly with soap and cold water. Urea Inhalation: Causes irritation to the respiratory tract. Symptoms may include coughing, shortness of breath. May be absorbed into the bloodstream with symptoms similar to ingestion. Ingestion: Causes irritation to the gastrointestinal tract. Symptoms may include nausea, vomiting and diarrhoea. May also cause headache, confusion and electrolyte depletion. Skin Contact: Causes irritation to skin. Symptoms include redness, itching, and pain. Eye Contact: Causes irritation, redness, and pain. Chronic Exposure: A study of 67 workers in an environment with high airborne concentrations of urea found a high incidence of protein metabolism disturbances, moderate emphysema, and chronic weight loss. ...read more.

Conclusion

*When the experiment is over repeat 3 times. Results Temperature (�C) 20 30 40 50 60 70 80 Volume of experiment 1 (cm�) 2.50 7.90 25.00 33.00 43.50 7.00 1.50 Volume of experiment 2 (cm�) 2.10 11.00 23.60 30.65 45.80 3.80 3.20 Volume of experiment 3 (cm�) 2.00 9.30 27.20 32.50 40.50 5.65 1.90 Average (cm�) 3.30 9.40 25.27 32.05 43.27 5.48 2.20 Analysis Temperature Average volume (cm�) 20 3.30 30 9.40 40 25.27 50 32.05 60 43.27 70 5.48 80 2.20 As from the graph and figures above, it shows that my prediction was right. As the temperature increases the enzyme produces more carbon dioxide gas until it gets to an optimum temperature of which in this case it is 60�C and as shown on the graph the carbon dioxide release rate decreased rapidly as the enzyme urease becomes denatured above this temperature and it will not work as effectively. This must mean that the rate of reaction increases, as the temperature increases until it gets to the optimum temperature and then it will start to decrease. This information can be shown by the graph I am going to produce below. Calculating the rate of reaction: 60secs � 5 = 300secs 3.30cm� � 300secs = 0.01 9.40cm� �300secs = 0.03 25.27cm� �300secs = 0.08 32.05cm� �300secs = 0.11 43.27cm� �300secs = 0.14 5.48cm� �300secs = 0.02 2.2 cm� �300secs = 0.01 Temperature (cm�) Rate of Reaction 20 0.01 30 0.03 40 0.08 50 0.11 60 0.14 70 0.02 80 0.01 BIOLOGY COURSEWORK - Enzymes 1 By Eunice Chidavaenzi *12P ...read more.

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