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Investigating the effect of temperature on Trypsin enzymes.

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Introduction

Investigating the Effect of Temperature on Trypsin Enzymes Plan Introduction I am going to be conducting an experiment to find out how temperature affects the rate at which trypsin hydrolyses protein found in milk. It is a biological catalyst, speeding up or slowing down the rate at which protein is broken down. I will test how quickly the trypsin works at a series of different temperatures ranging from 15� to 80� centigrade. Enzymes Enzymes are specialized proteins. They act as biological catalysts, which means they speed up reactions, but remain unchanged. Enzymes can be involved in reactions that build up material, or "synthesise" products, these are called anabolic reactions. They can also be involved in reactions that break down products, these are called catabolic reactions. They can, in theory, be used over and over again, but they do not last forever, due to wear and tear, and digestive enzymes go down with food; however, they can be reused. Enzymes work by the "lock and key hypothesis": Source: http://www.schoolscience.co.uk/content/5/chemistry/proteins/Protch6pg2.html Enzymes are affected by different conditions. * Temperature - enzymes work faster when they are warmer, due to kinetic theory. In fact, a 10�C rise in temperature doubles the rate of reaction. Though, as enzymes are made of protein and are delicate, they cannot withstand high temperatures, and the "active site" which is where the enzyme performs its job changes shape, if the enzyme is subjected to temperatures it can't handle. ...read more.

Middle

This means that if there is an anomalous result it is obvious, and I can still get an average from the other 2. I also will control the other two factors affecting enzyme rate of reaction: concentration and pH level, so I can focus on temperature. Using the same batch of powdered milk throughout the experiment will control PH. Concentration of both substrate and enzyme will also be controlled because I will use the same volume and batch of each. To achieve this as accurately as possible, I will use syringes to measure the quantities of each, keeping a separate syringe for the milk and one for the trypsin, this is to avoid potential contamination. Another problem experienced in the trial experiment was deciding what counted as "clear", as this is ambiguous. This was because although the solution was supposed to go clear, it never quite became absolutely colourless, thus I will have to control this by stopping the experiment once it gets to a certain shade. Hypothesis Enzymes work faster when it is warmer due to kinetic theory, that is, the particles have more energy, therefore they move faster so it is more likely a substrate will come into contact with an enzyme hence there will be more reactions per second. The accepted relationship between temperature and rate of reaction is that for every 100C temperature rise; the rate of reaction will double. ...read more.

Conclusion

However, there were some areas where more results would have been beneficial. Between 65o and 80o I should have taken at least one more results to determine where the enzyme finally denatures. Also, lower temperatures would have been useful to establish at how low temperatures trypsin works. Finally, more results around 55o would have been helpful to ascertain the peak temperature more accurately, as currently I can only assume that trypsin works best at 55o. From this experiment we can see that enzymes don't all act in the same way, and that trypsin doesn't act in the same way as some human enzymes. However, from the shape of the graph, that the change in rates of reaction is similar between enzymes, and that the "Q10 = 2" theory applies. Also, although I don't have enough results to confidently state the peak temperature, but I can say that it is approximately 55o and also that trypsin is denatured by 80o. To further the investigation into trypsin, I could investigate another factor that affects it, such as pH or concentration. I could use the same method, but use less or more of the substrate or the enzyme to see how concentration affects the rate of reaction. I could also keep concentration and temperature the same and affect the pH of the casein, which would determine the peak pH for the enzyme trypsin. By Daniel Lim ...read more.

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