temperature on the activity of liver

temperature on the occupation of colorfulIntroductionCatalase is a common enzyme found in upkeep and it acts as a protective mechanism for delicate biochemical machinery of cells. The enzyme catalyzes the exothermic hogwash of heat content bl to each one to piss and atomic upshot 8. 2 H2O2 2 H2O + O2 atomic number 1 henry henry peroxide is a by product produced by many living organisms during the process of metabolism. henry Peroxide is a very toxic center of attention (a power oxidizing agent) to cells and mustiness be broken down in influence to protect the cells from subsequent damage.AimThe aim of the taste is to investigate the proceeds of varying temperature has on the rate of enzyme catalyzed reaction. The foc utilize reaction is the decomposition of hydrogen peroxide with the enzyme catalyze. The presence of catalase muckle be demonstrated by dropping a dispirited piece of fresh coloured meander into quash hydrogen peroxide solution. In this auditio n, pieces of colored thread get out be ramble into different temperatures of peeing supply for 5 proceeding. After that the colorful wavers entrust be dictated into separate solutions of hydrogen peroxide and the measurement of oxygen botch up produced in a minute result be metrical using a assail syringe. HypothesisTemperature is a measurement of the ground take of hotness or coldness of a body or environment. more specifically, it is a measure of kinetic life force in a exemplar of matter. On a molecular level, temperature is the result of motion of particles which make up a substance. As the temperature increases, the motion akinly increases. The motion may be due to external energy applied to the particle of internal energy from the vibration of the particle. As temperature is increased, molecules deal increased bustling energy and reactions surrounded by them and the probability that the particles will collide with each other will also be greater, this change magnitude the rate of reaction. In chemical reactions, for every 10C rise in temperature, the rate of reaction most doubles. This office is known as the temperature coefficient of a chemical reaction. However in a enzyme catalyzed reaction the effect of temperature is more complex, for proteins change mannequin by heat. on that point are many factors that cease affect the structure of a protein such(prenominal) as temperature and ph. When a protein is exposed to heat, it serves the atoms to vibrate violently, breaking and disturbing bonds indoors the protein, therefore changing the chemical characteristics of the protein. I hypothesize that as the temperature of the pissing privy that the colorful interweave is exposed increases the nitty-gritty of oxygen flatulency liberated will also increase up. I believe that there will be an optimum temperature for the enzyme and going pass the optimum level will cause a drastic lessening in enzyme activity (less oxygen fuck up will be produced). Since catalase is found in virtually all living things, including humans, I predict that the optimum temperature for catalase will be VariablesIndependent VariableTemperature of peeing bath liver tissue is place In (C) Dependant VariableVolume of oxygen produced in a minute (ml/min)Controlled Variable C at one timentration of the Hydrogen Peroxide Volume of Hydrogen Peroxide Mass of liver tissueThe concentration of hydrogen peroxide must be kept constant because according to the Collision scheme proposed by Max Trautz and William Lewis in 1916 and 1918, increasing the concentration, increases the chances of particles hitting each other. The book of hydrogen peroxide should also be kept constant. Increasing the stack of hydrogen peroxide increase the substratum concentration and thus increasing the rate of reaction. Finally the mass of liver tissue should also be kept constant to try control the get of enzyme molecules present. Increasing the number of enzymes means there are more fighting(a) sites present and substrate molecules do non have to queue up for access to an active site. Ultimately increasing enzyme concentration mickle also result in an increase in rate of reaction therefore the mass of the liver tissue should also be controlled.EquipmentEquipmentQuantityNotesDigital Stop postdate1Thermometer1 0.5C Digital Balance to two decimal places1 0.01gConical Flask7250mlBeaker1500ml (for water bath) throttle valve Delivery Tube1Gas Syringe10.5ml give stand1Clamp1Boss 1 judicatory Mat2Safety Goggles1Deionized Water bottleful1Packet of Ice1Used for temperature beneath 30C Matches1Used to light Bunsen BurnerChemicals-Dilute Hydrogen Peroxide H2O21Concentration (2M)Volume (800ml)Safety NoteEye egis should be worn at all timesIf liquid gets into eye, oversupply the eye with a amiable running tap for 10 minutes and seek medical attentionIf hydrogen peroxide is spilt in the lab, describe it with mineral absorbent. Dilut e with water and wash liquid.Hydrogen peroxide should be stored in a dark brown bottle and care must be taken when removing the cap as it is possible that pressure may have built up inside it. MethodDraw up a sui dodge table or tables to record the results.Carefully ablation 7 pieces of cow liver tissue using a knife and a cutting mat.Weigh each piece of liver tissue carefully on the electric symmetricalness. Make sure each liver tissue weighs roughly around 0.5 grams.Place each liver tissue into a separate boiling tube and add 40ml of deionized water to each boiling tube once the liver tissue is situated at the bottom of the boiling tube.Place the heating mat on the table with the tripod on top of the heating mat. Gently place the gauze on the tripod. Once this is done, place the beaker on the tripod and slowly heat up the water with a Bunsen burner. Place a boiling tube with a liver tissue sample into the water and put a thermometer in the tube. Heat the beaker until liver samp le solution reaches 70C. Measure temperature of water with a thermometer.After that, carefully measure out 100ml of hydrogen peroxide with a measuring cylinder and transfer the solution to a 250ml conelike flask.Connect one end of the gas delivery tube to the gas syringe and the other to the conical flaskRemove the liver tissue from the boiling tube with a pair of tweezers and place it into the conical flask with the hydrogen peroxide.Quickly cork the conical flask once the liver tissue is dropped into the solution of hydrogen peroxide. Beginning timing the time once the liver tissue touches the hydrogen peroxide solution. Stop the stop arrest after 1 minute and record the aggregate of gas produced. register off the gas syringe.When the reading is taken, remove the cork and dispose of the hydrogen peroxide in the chemical waste container. Repeat the above step until data points from 10C to 70C are recorded.. For readings below 30C, cool the liver tissue sample with an ice bath .platResultsTable of ResultsVolume of Gas Produced in a Minute (ml)Temperature (C) effort 1 0.5mlTrial 2 0.5mlTrial 3 0.5mlAverage1 ml2032.033.035.0333040.036.041.0394045.047.050.0475054.052.054.0536063.060.065.0637043.037.040.040804.02.04.03Table 1.0 Raw DataTable 1.1 Qualitative ObservationsTemperature (C)Observations20Effervescene, gentle aglitter(p) in solution30Effervescene, gentle bubbling in solution40Greater effervescence, more bubbling in solution50Vigorous effervescence and bubbling60 fantastic effervescene, violent liberation of gas, bubbling in solution70Effervescene, gentle bubbling in solution80Bubbling in solutionGraph 1.0 Temperature and the Amount of Oxygen Liberated from Liver Tissue standard Graph 1.0 The graph above shows the relationship in the midst of the temperature of the water bath the liver tissue sample was put it and the amount of oxygen gas liberated from the sample after dropping it in dilute hydrogen peroxide in 1 minute. The graph clearing sho ws that as the temperature increases, the amount of gas also increases up to 60C. From 60C onwards, the amount of oxygen gas produced decreases drastically and there is a downward dispose of the curve.DiscussionFrom the data obtained, there is an increase of oxygen produced as the temperature of the water bath increases. This trend however only applies to the data points from 20-60C. At 70C however, there is a pregnant drop in the amount of oxygen gas produced and at 80C, the amount of oxygen gas produced is less than 5ml. From the graph, the relationship is clearly represented. Up to about 60C the amount of oxygen gas produced increases and ten-degree rise in temperature is accompanied by 6-7ml increases in oxygen gas produced. The amount of oxygen gas produced decrease at high temperatures as shown from 70-80C. So as the temperature rises, the amount of enzyme progressively decreases and the amount of gas produced is less. As a result of these two personal effects of heat on en zyme, there is an apparent temperature for an enzyme. Using the graph, the optimum temperature of catalase is approximately at 60C.The properties of a protein greatly depends on its three dimensional shape of the molecule. Exposure to heat causes the atoms to vibrate violently and this can cause bonds deep down the protein between different amino group pane of glass to break, resulting in a loss of the proteins biological properties. This is known as denaturation of a protein. Heating causes a proteins biological properties to change such as optical rotation, shape of active site and stick. The active site of the enzyme is what defines the enzyme. If the active site changes, the substrate molecules will no longer fit the active site of the enzyme. This is shown in the plot below. A proteins stability depends primary on the hydrophobic effects and to a lesser extent, by the interactions between polar residues and by other types of bonding. thither are four levels of protein struc ture each playing a significant in the stability of the protein. The primary structure of a protein is the sequence of amino acid residues attached by peptide linkages. Proteins differ in the variety, number and the ensnare if their constituent amino acids. Changes is just one specific amino acid in the sequence of a protein can alter a proteins property. The lower-ranking structure of a protein develops when parts of the polypeptide chain take up a particular shape. The most common shapes are either coiling to manakin helixes or into sheets. The tertiary structure of a protein is the distinctive and precise structure, crotchety to that specific protein. This is usually the result of further folding and interactions within the molecule. The shape is held together permanently by four different types of bonds hydrogen bonding between hydroxyl and ketone functional groups, Van der Waals forces between neighboring atoms, disulphide bonds between two cysteine side chains and ionic bonds between oppositely aerated ions. The stability of a protein is dependant of the balance of these three structures. Altering the balance of forces that brinytains the native conformation of the protein will lead to denaturation. ConclusionThe surmise put forth before the experiment was correct. As the temperature of the water bath that the liver tissue is exposed increases the amount of oxygen gas liberated will also increase up to a certain level. That level would be the optimum temperature of the enzyme and the optimum temperature of catalase deduced from the experiment is approximately 60C. Pass the optimum temperature, the amount of oxygen gas produced drastically decreases as the enzyme is starting to denature. At 80C, the enzyme is almost fully denatured and only a small amount of oxygen gas is produced. The literature value for the optimum temperature of catalase in human is about 37C which is the body temperature of a natural human being. The liver sample was taken fr om a cow and most(prenominal) cows and humans are able to regulate their own temperature. From this we can deduce that the optimum temperature of catalase for a cow should be roughly similar to a humans and somewhere around 37 -40C. The optimum temperature obtained from the experiment is 60C, this suggests that there may have been deviations or variations within the data. demarcation to ExperimentThere are several limitations to our experiment to the experiment. One of the main limitations was controlling the temperature of the water baths. The temperature began to drop gradually once the boil tubes were placed into the beaker. This may have caused deviation and variations within the data. Another limitation was that the experiment was only iterate 3 times and there may in time be room for anomalies and errors.EvaluationAs long as the temperature of the water baths were kept constant and the amount of gas produced was measured and read correctly, a good approximation of the opt imum temperature of catalase in a cow can be found. However there are several key improvements that can be made. When the liver tissues were cut a lot was wasted and some of it got stuck to the spatula and to the top of the test tube. To compact the problem in the near future, the liver tissues stuck onto spatula could be washed off using a tiny amount of water or shook gently to try remove some of the liver tissue. The method for collected the oxygen gas could also have been improved. Some gas was lost due during the time it took me to insert the bung into the test tube. To minimize the amount of gas lost, have a cooperator put the bung on for you while you drop the liver tissue into the measuring cylinder. The temperature of the water baths started to drop after a few minutes and this may have lead to some slight deviation within the data. In future make sure that the temperature of the water bath does not change too significantly. A slight modification would be to go forward t he beakers with a lid to prevent air from escaping. Another problem that arose during the experiment was keeping the temperatures of the water baths constant during tests and between trials. The problem was that different temperatures of liver samples were tested and it wouldnt be feasible to use a electronically controlled water bath to perform each test. The result was that a Bunsen burner was used but this creates several problems. It was extremely difficult to get exact temperatures and reserve them throughout the experiment. In future, the use of a heating plate would be more practical so as to prevent drastic changes in temperatures. To improve the experiment it may be fall in to use a solution of catalase rather than a liver as a tooth root of catalase. By using a solution, the concentration and volume could be controlled and would be the whole experiment into a fairer test. Overall the results obtained have helped support the hypothesis put forth before the experiment. Fi nally the experiment could have been repeated a few more times so that a better average could be drawn from the data, resulting in a more good and accurate conclusion.ReferencesClegg, C. J. (2007). Biology for the IB Diploma. London Hodder Murray. Ophardt, C. E. (2003). Denaturation of Proteins. Retrieved January 3, 2009, from Elnhurst College Web site http//www.elmhurst.edu/chm/vchembook/568denaturation.html Roberts, M. B. (1986). Biology a functional approach (4th ed.). Canada Nelson. Voet, D. J., Voet, J. G., Pratt, C. W. (2008). Principles of Biochemistry. NJ Jon Wiley Sons, Inc.