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The Effect of Temperature on the Respiration of Yeast Planning Aim For my coursework I am going to investigate the effect of temperature

on the rate of respiration of yeast. My aim is to find out if there is any correlation between the rate of respiration of glucose by yeast and the mixture's temperature. I shall do this by conducting a simple safe experiment, which will involve the timing of the yeasts respiration, when mixed in a glucose solution. This shall be done at various temperatures. To obtain the best range of values to use in my final experiment, I shall conduct a preliminary experiment on the concentration of glucose solution. In this way I will also be able to increase the accuracy of the final experiment by seeing if the effect of glucose solution on yeasts respiration will have any effect on the main experiment. Variables In my main experiment, I am going to use temperature as the changing variable. Here is a list of variables that I am keeping constant. Amount of yeast Concentration of glucose solution Amount of glucose solution Hypothesis, Theory and Scientific Knowledge Temperature There are many ideas to suggest that the change in temperature will cause an increase of respiration in yeast. Yeast is a single celled fungus made up mostly of protein, which is used in fermentation. Fermentation is the breakdown of sugars by bacteria and yeast using a method of respiration without oxygen (anaerobic respiration). It involves a culture of yeast and a solution of sugar, producing ethanol and carbon dioxide with the aid of the enzymes. The alcohol produced has been used in making wines and beers and the carbon dioxide produced has been used in baking as it gets trapped in the dough and causes it to rise. Enzymes are catalysts that speed up reactions; they are made from protein and are specific as to which substrate they work on. For example a zymase-complex enzyme will only bind with a glucose molecule to produce the ferments carbon dioxide and alcohol.

Yeast has to make energy, stored as ATP to carry out all cellular functions. To do this they respire. They can respire both aerobically (when there is plenty of oxygen and the cells reproduce rapidly), or, where oxygen is short, they can respire anaerobically; in this process they are called partial anaerobes. This is because less energy is released as the glucose sugar is only partially broken down, but still keeps the yeast alive. In my experiment the yeast is respiring by anaerobic respiration. Here is the equation for anaerobic respiration: Enzymes in cytoplasm (Zymase complex) Glucose -----------------> ethanol + carbon dioxide +energy C6H12O6 2C2H5OH CO2 The Kinetic theory states that, with an increase in temperature, the rate of reactions will increase. This is due to the increase of speed of the particles, brought about by the extra energy given to them by heat. The faster particles will bring about more particle collisions and so the reaction will take place faster. Enzymes are sensitive to temperature changes up until a certain temperature and will increase in their activity up to this point. The reactions that take place in the enzymes will be quicker and so will create more of their products. In general, it has been said that there is a doubling of the rate of reaction for every 10C rise this is called the 'Q10=2' theory. This should be evident when the concentrations of the yeast and glucose solution are kept the same also. But once you reach a certain temperature the rate of respiration slows down and drops. This happens because; all the enzymes are made up of protein chains of amino acids. They exist in the form of a helix structure with hydrogen bonds holding them together. When heat is applied to the enzyme, energy is given off. The active enzyme cell deforms and the hydrogen bonds break, denaturing the yeast enzyme. It would not be able to function as usual, and this process is irreversible. This process is therefore called denaturising. The optimum temperature in which yeast enzyme work best is around 37C, below this the rate of reaction is slow and above 45C the yeast enzyme would denature. The analogy of denaturing is to compare a key to a keyhole. If the keyhole has changed, the same key will not fit in any more, and the lock will not be unlocked. The same thing happens when the yeast enzyme is denatured, therefore fermentation cannot continue after this has occurred. Firstly though, I am going to carry out the preliminary experiment to

see whether the concentration of glucose solution will affect my main experiment. Preliminarily Work

Prediction ========== As the concentration of the glucose solution goes up the more bubbles of gas will be produced. Therefore there will be an increase in yeast enzyme respiration. This will be because the sugar molecules will be more abundant as the concentration increases, so the probability that the enzymes come into contact with the molecules is higher and therefore more energy will be produced in the form of anaerobic respiration more often. Diagram Results Experiment 1 Concentration of Glucose in Water Total no. of bubbles Time 1 2 3 1st Minute 13 14 9 36 2nd minute 10 7

4 21 3rd Minute 4 7 4 15 3rd Minute 3 8 3 14 Average 7.5 9 5 Experiment 2 Concentration of Glucose in Water Total no. Of bubbles Time 1 2 3 1st Minute 18 25

26 69 2nd minute 3 9 10 22 3rd Minute 2 5 9 16 3rd Minute 3 4 12 19 Average 6.5 10.75 14.25 Conclusion What I could notice from my graphs is that the rate increases proportionally as concentration increases in the second result graph proving that my prediction was correct but not in my first graph. This I am putting down to an anomalous result and human error when the results where being collected and therefore I am not including this as my main conclusion .The rate of reaction depended on the surrounding temperature and the concentration of the solution, therefore the more concentrated the solution is the faster the rate. This therefore shows

that during my main experiment, in which I am going to be investigating temperature, I must make sure that my set glucose solution is accurate and does not change during the experiment as this could affect the out coming results leading them to be inaccurate and therefore invalid. Main Experiment Prediction With reference to my theory, I predict that the rate and speed of respiration of glucose solution by yeast will increase with temperature rise up until a certain point where the enzyme used and secreted by the yeast will become denatured and cease to function (denature), reducing the rate significantly. This is explained through Kinetic theory, yeast respiration and the nature of enzymes. Main Experiment Method Apparatus: Syringe, Fresh yeast, Glucose solution, Boss head, Glass Rod, Spatula, 10cm measuring cylinder, Stop clock, Weighing scales, Thermometer, Water Bath. Accuracy TEMPERATURE Temperature of the experiment will have a great affect on the results as explained by kinetic theory. Temperature will affect the rate of yeast respiration. I shall keep the temperature of the water bath under control by using a thermometer and checking it constantly. I shall also keep swirling the thermometer to keep the heat distributed.

AMOUNT OF YEAST The amount of yeast is crucial, more yeast means more glucose will be respired and more products created. An imbalance will upset the results. The amount of yeast will be weighed out on an accurate balance each time. I have decided to use 0.5 grams of yeast for my experiment as accuracy is very important in obtaining the correct and most reliable results. Also it is important to keep the constant variables constant. AMOUNT OF GLUCOSE SOLUTION The amount of glucose solution will affect the results, as my preliminary work has proven. More glucose solution means that there are potentially more products, which would make the results more inaccurate and potentially make an unfair experiment. The volume of glucose solution I have chosen to use is 3% as the results of my preliminary work show that the rate of respiration of the yeast was at its highest throughout the experiment at this concentration. It will be kept constant by using a measuring cylinder at each preparation. Procedure Using the spatula take a fresh piece of yeast and weigh it to 0.5 grams. Put 5cmof the 5% glucose solution into the measuring cylinder. Mix the glucose solution and the yeast together gradually until it is all mixed together. Draw up the 5cm yeast and glucose solution mixture into the syringe and pull out the plunger to the graduation mark. Rotate the yeast /glucose solution mixture away from the nozzle (in order to prevent spillage). Using the water bath take the temperature using the thermometer making sure that it is at the right accurate temperature. Place the syringe into the water bath and place the boss head on top of the syringe to make sure that it is fully immersed and stays down. Count the number of bubbles produced by the yeast/glucose solution mixture in one minute using the stop clock. Remove and wash out the apparatus. Repeat this same method but change the temperature according to the temperature range. (20to 60) going up by 5each time in repeating the Repeat the overall experiment twice to get a wide range of results. This will improve the accuracy and will also enable me to take the average of the results and produce a 1/time Graph.

Safety It is important that I keep into consideration the safety of the experiment: Choose Reasonable temperatures to experiment with; e.g. If the temperature is too high (say, 100) it would be too dangerous to work with. Make sure that all electrical equipment is away from any water e.g. the water bath. Wearing safety goggles when necessary. Don't carry water across the room because if you spill it, it could be hazardous and could cause an accident. Method Diagram I would collect my results in a table like this: Temperature C Amount of bubbles produced in 1 minute Result 1 Amount of bubbles produced in 1 minute Result 2 Average amount of bubbles produced in 1 minute 20 25 30 35 40 45 50 55 60 65 70 Due to difficulties I was unable to do my experiment and our class

conducted a class method: CLASS METHOD STUDENT NUMBER: MATERIALS: syringe 18 x 20 cm3 fresh yeast approx 60g (18 X 0.5g X 3 + extra for extra readings and mistakes) glucose solutions 5% (18 x 5 x 3 = 270 cm3) water baths 1 plastic tray and 5 water baths (all with "blue" thermometers) 100 cm3 beakers; x2 measuring cylinders PROCEDURE: 1 Three students will be placed at each "temperature station". There will be six stations around the lab. 2 Weigh out 0.5g of fresh yeast in a small beaker. Put 5cm3 of 5% glucose solution into the measuring cylinder. Add some of the 5% glucose solution to the beaker and mix the yeast and glucose together. Keep on adding the glucose until all the 5 cm3 is used up. 3 Draw up the 5 cm3 yeast/glucose mixture into the syringe. Pull out the plunger to the final graduated mark. Rotate the syringe so that the yeast/glucose mixture is away from the nozzle. 4 Place the syringe under water, in the water bath to which you have been assigned (room temperature should be approx. 20oC -measure this if you are assigned this "station". Check that each water bath is at the correct temperature, using a thermometer, at all of the other stations). Place the boss head on top of the syringe to hold it down. Collect the amount of gas produced in a small measuring cylinderremember to start at a graduation mark on the inverted cylinder. 5 Remove and wash out the apparatus. 6 Repeat steps 2 to 5 counting and recording 3 times. Collect the results from the rest of the class, enter them in the table provided and work out the average volume of gas produced. Water temp

(oC) Amount of gas produced per minute cm3 Average volume of gas produced cm3 1 2 3 4 5 6 7 8 9 room temp 0.13 0.02 0 0.03 0.07 0.07 0.03 0.03 0.02 30 0.43

0.25 0.23 0.35 0.25 0.2 0.35 0.18 0.1 40 0.6 0.4 0.2 1.0 0.4 0.43 0.2 0.08 0.05 50 2.2 0.5 0.53 2.6 1.1 1.0 2.3

0.8 0.6 60 2.0 1.2 0.5 1.7 0.83 0.5 2 0.83 0.5 70 4.7 1.5 1.08 2.5 1.58 1.33 Analysis The rate of reaction table can be seen in the obtaining evidence class method. Due to complications and that there was a limited supply of equipment we conducted a class method of which we shared the results. From looking at my results and graphs you can see that as the temperature increases there is a rise in the amount of bubbles of carbon dioxide produced in respiration and therefore backing up my hypothesis and prediction with the resulting outcome in that the results of the graph are inversely proportional. The results are explained by the theories of enzyme denaturising with the lock and key and kinetics theories. Where these meet is when kinetic theory states that an increase in temperature will equal more particle collisions

between the glucose solution and yeast and so a faster rate of reaction will take place; and in enzyme denaturing where the enzyme is sensitive to heat, and about a certain temperature, the active enzyme will begin denaturing, so slowing and eventually stopping the reaction. This will give an area where the rate of respiration drops off and goes to nothing instead of a precise 'cut-off' point. These did not however both apply to the experiment as the enzymes continued to function after the yeasts maximum temperature for normal enzyme activity. The reason for this I believe is that the temperature I measured was that of the water bath, the surrounding temperature, and not that of the actual glucose solution. Therefore I had no way of knowing if the glucose solution was in fact at the same temperature as the water bath .If the glucose solution had not reached that of its maximum temperature than it would not have denatured and this would therefore make my theory correct. But instead the heat would have acted as a catalyst on the yeast and glucose solution. I did find an anomalous result when looking at the results table. This could be explained by the inaccuracy of results and that the reaction could not be totally accurately controlled with the apparatus used. Conclusion I can conclude form my graph that my prediction and hypothesis was correct that there is definitely a positive correlation between the rate of respiration of yeast in a glucose solution and temperature and that as temperature increases the rate of respiration increases. Evaluation To make sure that the results were as reliable as we could make them, we calculated the mean of 9 results at each temperature interval except on the last temperature when we only managed to obtain 6 results. The fact that we only managed to obtain 6 results at 70c will have affected the results and the graph I have plotted therefore I Know that there will be an error in the graph and is probably the cause of my overall slightly higher result at 70c. Although the results seemed quite accurate I believe that mistakes could easily have been made and that the results are not as accurate as they could have been. This is because each temperature was measured by a different member in the class group and that each member could have made a slight error either in calculations or by human error (e.g. counting the amount of gas produced).If this was the case then results would be untrue and therefore our group results combined together would be formed on the trust and believe that each member had completed the experiment accurately and correctly which would mean the result were extremely unreliably. As a group we took all precautions to make the apparatus used to be

reliable and so I think the slight unreliability was caused by the preparation of the solution and the 'unpredictability' of how the reaction went that came with it. To obtain more reliable results and if I were to conduct the experiment separately I would want complete continuity with preparations, maybe arranging 'sets' of substances to create several solutions of glucose beforehand and adding the yeast separately at a set time but not actually activating the yeast until necessary so as to prevent any solutions getting a 'head start' over the others. This would ensure that all the preparations are the same and would give continuity. I would want to be more strict and thorough with preparing solutions and mixing them up. I would want each one to be thoroughly acclimatised to the surroundings and had the same amount of glucose and the same activating and mixing time. This would help give more reliable results throughout. Another alteration to my project would be changing the way in which I collected the gas. This would increase the accuracy of the results if they had a human error. This is because the measuring cylinder we used may not have been filled at a correct graduation mark and therefore the results would have been untrue as there would therefore have been human error of the results + or- a graduation mark. The anomalous result produced at 50c was at first a slight concern as it was so very much higher than the results at 60c.At first I had assumed that this was because the yeast enzymes had denatured but as the result at 70c was very high I ruled out that possibility. This therefore means that the high reading at 50c must have either have been of a group calculation error or that of a human error. If I were to further investigate this experiment and my results, I would probably want to calculate the point where the enzymes begin to denature for respiration in yeast. I could also examine the change in rate between the intervals to determine validity and continuity. I would also have taken more results at each temperature interval as to increase the depth and accuracy of the results and therefore would help in achieving a better graph with a stronger correlation.