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Investigation in to the effect of temperature on the activity on the enzyme Trypsin.

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Introduction

ANDREW MESSIOS 10P MR.ARMON - SET: TRIPLE 2 TRYPSIN AIM To investigate the effect of temperature on the activity on the enzyme Trypsin. INTRODUCTION: An enzyme is a biological catalyst which means it speeds up the reaction without itself being used up. Enzymes are vital in the digestive systems as they break down substrates (e.g. protein) into substrates (e.g. peptides and Amino Acids) to be absorbed into the blood. However even though enzymes are never used up, the body must still constantly produce them because they break down with the food and are absorbed. This is because enzymes are made of protein structures. All enzymes have active sites. Active sites are dents in the surface of the enzyme where the substrates fit into and are broken down in. Each molecule can only break down one substrate at a time (lock and key theory, see below). Trypsin is one of the many different types of enzymes, each with its own specific function. It is often referred to as a protease. Trypsin's function is to break down proteins into their component peptides and amino acids so it can be absorbed into the blood and used by the body for growth and repair. Trypsin is produced in the pancreas and continues the process of digestion (begun in the stomach) in the small intestine where a slightly alkaline environment (about pH 8) promotes its optimum enzymatic activity. The film which will be tested is composed of 3 different layers. The top layer is transparent acetate. The middle layer is a thin coating of gelatine to hold the acetate to the third layer, the Silver Nitrate. The gelatine is the key ingredient to this experiment as it is made of protein and will be broken down by the Trypsin to show the film as clear. The film appears black before tested because the gelatine and acetate are transparent and the Silver Nitrate is black. ...read more.

Middle

This average should ensure more accurate results by confirming that a single measurement wasn't an off chance or an anomaly. The 3 test tubes for the same temperature will be measured at the same time to economise the time we have and to make sure that that particular temperature had the same variables and invariables for each test tube. Another minor change made is the way we will check if the film has gone clear. Instead of taking the film out of the test tube, we will now take the test tube out of the water bath. This is done because each time the film is taken out the force which the film is under when it leaves the water could remove some of the gelatine, slightly corrupting the results. Our new method will not significantly change the temperature of the contents of the test tube as the tube will only leave the water bath for a very short period of time. Apart from the above changes to the experiment, the rest of the procedure will exactly follow what is stated in the preliminary method. RESULTS: Temperature (�C) Individual Times (s) Average Time (s) Rate (s) 0 360.00 (CUT OFF) 360.00 (CUT OFF) 360.00 (CUT OFF) 360.00 (CUT OFF) 2.78 (CUT OFF) 20 270.00 340.00 335.00 315.00 3.17 40 70.00 70.00 120.00 86.67 11.54 60 90.00 150.00 130.00 78.33 12.76 80 360.00 (CUT OFF) 360.00 (CUT OFF) 360.00 (CUT OFF) 360.00 (CUT OFF) 2.78 (CUT OFF) ANALYSIS OF GRAPHS: The following analysis will be on my rate of reaction graph in the final experiment: The first result in the graph is a cut off time at 0�C (rate of reaction time is 2.78 because in the calculations for the rate we used 360s as the time). The result was cut off because of the cold temperatures the Trypsin enzymes were put in to break down the gelatine in the film. ...read more.

Conclusion

When the colourimeter has sensed the film has gone completely clear, the time will stop. The results will be far more accurate, depending on the accuracy of the watch (usually to a 100th of a second). There were another number of possible reliability and accuracy issues in the execution of the experiment itself. For instance the 20�C temperature was not measured in a water bath, but in the room itself. This was because 20�C is roughly room temperature. The fact however is that room temperature is usually 23�C and with the amount of heat produced from various water baths in the room and external hear sources such as the sun through the windows the room temperature would be almost certainly higher. So to improve reliability with this result a water bath could have been utilised as with the other temperatures. The water baths themselves however may not have been the exact temperature they should have been. Cold test tubes entering the water may have slightly decreased the temperature inside the water bath which would affect all results. The consequences of this are so miniscule however that I do not believe they would have had a huge impact on the results. And even so there isn't really a way round it because if you pre warm the test tubes it may increase the temperature of the water baths which is equally as bad. A last problem I found with the experiment was the measuring of 0�C which took place in a bucket of ice. Over time the ice used would start to melt because it would become warmer which means that you may not be measuring at 0�C but at 0.8�C or 1�C which would slightly alter results. Again however, the consequences of this would be very minimal and not worth noticing or correcting. Because of the scale of this experiment and the basic equipment provided I believe we maximised how accurate and reliable our results could be. This experiment was very accurate for the apparatus we used which is fairly simple and provided us with the overall correct results. ...read more.

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