Experiment-Effect of Temperature on the Action of Amylase Enzyme
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ï»¿________________ ________________ Mohamad Zulhilmi bin Azmi Biology HL EXPERIMENT : The Effect of Temperature on the Action of Amylase Enzyme INTRODUCTION Enzymes are biological catalyses, most of them are protein. A catalyst speeds up the rate of a chemical reaction without itself being changed at the end of the reaction. A few properties of protein are â enzymes are extremely efficient, extremely specific, can be denatured by heat (temperature) and are affected by pH. Amylase is an enzyme that is involved in the human digestive process. Found in both the human pancreas and the human saliva, amylase breaks down starch into sugar so that large molecules can be easily digested. Like all enzymes, amylase must be kept in a certain condition in order to function properly. In this experiment, the effect of temperature on the rate of reaction of amylase will be investigated with the use of starch and iodine. When starch is mixed with iodine, the coils of beta amylase molecules found in starch trap iodine, causing the mixture to turn into a shade of blue-black. When starch is broken down into glucose, however, the monosaccharide does not react with iodine. Therefore, glucose does not change color even when itâs mixed with iodine. Correspondingly, when drops of amylase are inputted into a blue-black mixture of starch and iodine, the starch molecules will be broken down into glucose molecules, causing the mixture to turn colorless.
5. Stop the reaction after exactly 10 min by adding 5 mL of HCl. 6. Add 0.5 mL of the above mixture to 5 mL iodine solution to develop color. Shake and mix. The solution should turn deep blue if there is any residual, unconverted starch present in the solution. The solution is brown-red for partially degraded starch, while it is clear for totally degraded starch. 7. Measure the absorbance with a spectrophotometer at 620 nm. RESULTS Quantitative data: Temperature/°C (±0.05°C) Rate of absorbance(%T) Rate of enzyme activity(Abs) 15°C 4 0.682 25°C 5 0.978 35°C 10 1.468 45°C 6 1.264 55°C 5 1.117 65°C 5 1.065 75°C 5 0.901 85°C 3 0.679 Qualitative data: Temperature/°C (±0.05°C) Color of the mixture after mixed with iodine Inference 15°C Deep blue Presence of unconverted starch 25°C Brown-red Starch was partially broken down 35°C Brown-red Starch was partially degraded 45°C Brown-red Only part of the starch was broken down 55°C Deep blue Presence of unconverted starch 65°C Deep blue Presence of unconverted starch 75°C Deep blue Presence of unconverted starch 85°C Deep blue All starch was not broken down Processed data: CONCLUSION The purpose of this experiment was to determine the effect of different temperatures on the reaction of amylase. The rate of absorbance and enzyme activity which was calculated using the spectrophotometer are what determine the effect of the different temperatures.
For example, because the solutions were very sensitive during reactions, a harder stir at the solution may have speeded up the reaction. SUGGESTED IMPROVEMENTS Numerous ways can be implemented in order to improve the results of the experiment. Firstly, the time intervals for mixing the starch and amylase solution as well as the addition of hydrochloric acid into the mixture should be standardized and short for an instant 10 seconds. The period of 10 seconds in between each interval is suitable and short enough in order to find out more accurate results about the rate of amylase enzyme activity. Before entering the cuvette into the spectrophotometer, the mixture of starch, amylase and iodine solution should be diluted first with distilled water. The justification will be easier for the penetration of wavelength so that more accurate results with more diversity will be produced. Hence, we can see clearer distinctions of the rate of absorbance and amylase enzyme activity in every temperature. Moreover, the experiment must undergo repetition at least 3 times. With more trials, mean of the data can be calculated with higher accuracy and consistency. Therefore, the results will be more reliable because we use the average value of the data instead of depending on only a single trial. Lastly, we can set a constant number for inserting and stirring the solution using the glass rod. This would add more uniformity on the reading of the rate of absorbance and amylase enzyme activity in every temperature.
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