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A Comparison into how temperature effects the rate of reaction of Bacterial amylase and Fungal amylase.

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Introduction

A Comparison into how temperature effects the rate of reaction of Bacterial amylase and Fungal amylase. 'Amylases hydrolyse glycosidic bonds in polysaccharides such as starch...to maltose.'p39 molecules and cells-Nelson advanced science. They are found in two forms - ? and ? amylase. Both work by hydrolysis - adding one molecule of water across the glycosidic link. Hypothesis My hypothesis is that bacterial amylase will be more resistant to denaturation at temperature exposure (above 40 c) than fungal amylase. Justification Enzymes, being proteins, are sensitive to environmental change. Temperature (and pH) can affect the activity of a molecule, as they can change the shape of the enzyme molecule. As temperature of molecules in solution increases, the more kinetic energy both enzyme and substrate molecules contain, therefore colliding more energetically and frequently, thus increasing rate of reaction. However, after the initial rise in the rate of the reaction, as temperature continues to increase, due to enzymes being made up of protein, they are adversely affected by high temperatures, and often above 45 c many enzymes are denatured. However, temperature increase, also affects how stable the enzyme molecule is, as precise shape of the active site is essential for catalytic activity. Therefore, a slight change in the shape of the active site, would result in it not being able to combine with its substrate, thus affecting rate of reaction, concluding in the enzyme being de-natured - when it loses its catalytic properties, and is unable to combine with its substrate, due to irreversible damage of the tertiary structure. ...read more.

Middle

1.Measure 2cm bacterial amylase solution into a clean test tube, 2.Measure 2cm of 1 % starch solution into another clean test tube. 3.Place test tube containing enzyme solution into a water bath, of temperature exposure 50 c, and test tube containing starch solution in a water bath into a water bath of control 40 c, and leave for 10minutes - place a thermometer in the tubes, to ensure correct temperature is maintained. 4.Using a pipette, place a drop of iodine into each dimple in a dimple tray 5.At exactly 10minutes, remove the enzyme from temperature exposure, and leave in control water bath ( 40 c) for approx 1min - or until the enzyme has cooled to control temperature. 6. When enzyme has cooled to control, mix it with the substrate - starch solution. 7. After1 minute, using a pipette, spot one drop of the solution, into a pit containing one drop of iodine solution on the spotting tile, and record colour change. 8 .Repeat step 7 every minute until denaturisation occurs. 9 .Repeat steps 1-8, but substitute the temperatures of the water baths for; 60 c, 70 c,and 80 c respectively. 10. Repeat steps 1-9, but substitute bacterial amylase for fungal amylase. 11. Repeat steps 1-10 at least two times, an order to eliminate any anomalous results. Variables As a means to ensure that the test is fair, I shall only vary two factors (at different stages, not simultaneously.) ...read more.

Conclusion

80 c, as minor burns can be incurred. If burns incurred, skin should be put immediately under cold running water, and medical advice sought. I must also ensure that any shards of broken glass are cleared away thoroughly, so as to eliminate unessessary injury, and that they are placed in a separate disposal unit to ordinary waste product. I must also wear protective clothing at all times so that I do not either contaminate my solutions, or wreck my outer garments if spillages do occur, and I must wear safety goggles, in order to protect my eyes. All solutions must be disposed of carefully - i.e down the sink (as the solutions that I will be working with are not highly toxic) and then the sink must be rinsed thoroughly. All apparatus must also be washed and sterilised, as the majority will be used in future experiments - so as to avoid any possibility of future cross-contamination. In the event of an outbreak of fire, all substances must be left inside the building, and caution must be taken to place them so that they are not at risk of being spilt, then sharp exit must be ordered through the nearest fire exit. Organisation must also be taken into account, and ensured that all test tubes and containers are clearly labelled, and when finished stored correctly, so again to eliminate any spillages etc.. It must also be ensured that long hair is securely tied back, and if ties or loose clothing is worn, that they too are securely fixed. ...read more.

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