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We are going to find out how different enzymes (in this case, trypsin) concentration in a solution would affect the rate of which solute (Casein) is broken down.

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Introduction

Enzyme concentration and enzyme activity For this investigation, we are going to find out how different enzymes (in this case, trypsin) concentration in a solution would affect the rate of which solute (Casein) is broken down. I think enzyme concentration would definitely have an effect on the rate of reaction. My hypothesis is that increasing the concentration of trypsin would also increase the rate of catalysation of casein. Enzyme (a globular protein) is used to catalyse specific biological reactions by lowering the activation energy and they are left unchanged by the end of the reaction, for example trypsin is responsible for catalysing the break down of casein (a protein typically found in milk - it is what makes it ?opaque?) into smaller molecules which the body can absorb. This means the reaction can occur in a faster pace. This is important since without enzymes the molecule cannot break down fast enough on its own to provide us with constant nutrients. The reaction happens with an enzyme-substrate complex forming between trypsin and casein.The formation of this complex is explained by the induced- fit hypothesis,where the enzyme has a distinct and flexible active site, as oppose to a rigid one suggested in the lock-and-key model. Once the casein enters and attaches to the active site of trypsin, the shape of its site will modify around the substrate to form the active complex, once the reaction is complete, the now broken down substrate will leave the active site and the enzyme reverts to its previous, unmodified form. ...read more.

Middle

Our team has decided to take the absorbancy reading every minute for 10 minutes since it seemed like an appropriate time interval, 30 seconds seems to rushed and we may not be able to see a clear pattern of absorbancy if we take the reading more than 1 minute apart. Furthermore, we?ve decided to set the observing period as 10 minutes because rate of reaction seems to start to even off by that time and so we would not need to observe any longer. We then discarded the content in the cuvette and rinsed it with distilled water repeated all the steps from adding 2cm3 of casein for each of the remain concentration of trypsin instead of using distilled water. We had to rinse the cuvette to make sure the solution left over would not affect the result of our next concentration. Apart from all the precaution we?ve set up in our investigation, there are still errors made as shown in our original result (very inaccurate, the values increased rapidly instead of decreasing), we could have taken more care to improve our method. These improvements will be mentioned and explained later on. (The results on the next page are ones we obtained from our class since our own results are too inaccurate to use.) The class result: The results shows there is definitely a decrease in absorbancy as we decrease the trypsin concentration at a certain time interval. ...read more.

Conclusion

We used precautions to try and reduce the amounts of errors made. However, in the end the calorimeter we used (to produce our set of result) seemed to be faulty, therefore there wasn?t really anything we could?ve done to avoid it apart form early detection (of faulty apparatus) by performing a preliminary trial. Overall I would say this is not a very successful investigation within our team since looking at our own result, it did not prove my hypothesis. It was only when looking at the class results was I able to make correct deduction and analysis. Risk assessment Hazard Risk Precautions Test tubes * Breaking it and hurting yourself / others * Use test tube rack * Remove broken glass immediately * Avoid cutting yourself when removing shards. Electrical appliances (water baths) * Electric shock * Keep water away from plugs * Do not use wet hands to plug in/ unplug Chemicals / solutions used * Have an allergic reaction to trypsin or casein * Be aware that they can be an allergen * Avoid contact as much as possible * Rinse immediate if it dropped on your skin Many people crowded at one place (near water bath) * People tripping/ falling and hurting themselves * Be careful when taking out chemicals * Avoid collision with other people. * Do not run or walk carelessly Reference/ image sources pH graph http://alevelnotes.com/Factors-affecting-Enzyme-Activity/146 Enzyme - Substrate complex diagram http://www.hsc.csu.edu.au/biology/core/balance/9_2_1/921net.html Enzyme concentration graph http://alevelnotes.com/Factors-affecting-Enzyme-Activity/146 ...read more.

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