Enzymes

Cristian Sanchez

Dr. Fazio

Hirum Ramos 10/15/09

Abstract: Lab number six is based on how enzymes may impact a reaction taking into account the several surroundings changing. Such as: temperature, concentration, and ph. An enzyme is a protein that catalyzes reactions without being consumed into it. What that means is that all enzymes are proteins that speed up reactions with the capability of doing it over and over again. The hypothesis we came up with was that: enzymes are proteins, so this means they should react like them by speeding up a reaction when met with appropriate amounts of concentration, ph, and temperature. Now we made this conclusion because of the information given in the lab report. For the concentration exercise it states that every reaction has a limit to how fast it can go, so after a certain point the reaction will not be able to go faster because all of the active sites will be occupied. The same goes for temperature, when adjusting the temperature to a reaction there will be only one prime degree to which the reaction will perform the fastest. When the temperature is either too high or too low the enzyme molecules begin to denature, which means to destabilize the structure of the protein. Ultimately the same goes for ph, there is only that one sweet spot level of ph that seems to get the reaction rate the highest. If the ph is too low or too high the hydrogen bonding and the structure of the enzyme are altered concluding with a slower reaction. Throughout the whole experiment catalase is the enzyme that is tested with all the various situations. Catalase is an enzyme that speeds up the breakdown of the toxic chemical hydrogen peroxide to water and hydrogen. I believe that all the results were significant, I feel this way because I imagined different results for tube one in exercise two and both tubes three in exercise's three and four. I though that the bubble height in tube one- exercise two would've been lower than it was because of the low temperature. One could believe that the reaction rate would be a lot slower than that of tube two (37D/C), but there was not that much of a difference. As for tube three- exercise three I felt that the bubble height would be lower than that of tube two, but I guess this tells us a higher concentration level is allowed into this reaction to catalyze it. And last with tube three exercise four, I expected tube three(70mm) to be close to tube one(5mm), but it was not, so this leaves us with the conclusion that the lower end of the ph is less effective then the higher end of the ph scale(bases). Introduction: Enzymes and their capabilities of catalyzing are actions that take place in human bodies everyday. Without enzymes digestion would be very different and it most likely would take a lot longer. For example, people that are lactose in tolerant can't break down lactose because their body is unable to produce lactase. Those that do produce lactase are able to ingest and and digest lactose products properly and don't have to worry about anything. Enzymes are what's being experimented in these exercise's. The main focus of this lab is to prove that enzymes react exactly like proteins, because that's what they are. So with that being stated, enzymes should speed up the reaction rate to the fastest possible speed when their concentration, temperature, and ph are all in there optimum levels. In addition, the reaction rate should be slowed down when the enzymes are denatured or in low reaction environments. Materials: Catalase hydrogen peroxide ruler wax pencil Three test tubes for each individual exercise H20 Sucrose Refrigerator Incubator Hot plate with a beaker and water in it

ph3, ph7, and ph 11 solutions Rack for holding test tubes stopwatch Method: *All height measurements are done in Millimeters Exercise One: Catalase Activity One may begin with labeling three test tubes at the 1cm and 5cm level with a wax pencil. Then you fill the first test tube with catalase up to the 1cm, after that you fill the rest with H2O2 up to the 5cm mark. You mix and stir, wait about 20 seconds for bubbles to appear, then you measure and record how high the bubbles went from the liquid. Second, you fill the second test tube with water up to the 1cm mark, and then you fill rest with H2O2 up to the 5cm mark. Next you mix and stir, wait 20 seconds, and then measure how high the bubbles got. With the last test tube you fill catalase up to the 1cm mark, after that you fill the remaining space with sucrose up to the 5cm mark. Concluding with the mixing, stirring, and measuring of the bubble height recording all the data of the test tubes. Exercise Two: Effect of temperature on enzyme activity One may begin once again with labeling three test tubes at the 1cm and 5cm level with a wax pencil. Then you may begin with filling all the test tubes up to the 1cm mark with catalase. Next you place the fist test tube in the refrigerator (4D/C), the second test tube in a incubator (37D/C), and the third test tube is place in boiling water (100D/C). After leaving all those in their new environments for fifteen minutes they are removed and all filled with H2O2 up to the 5cm mark. Last mix and stir, let it sit for about 20 seconds, then record the height measurements of the bubbles. Exercise Three: Effect of concentration on enzyme activity One may begin by gathering three test tubes labeling one at the 1cm and 5cm level, the second test tube at the 2cm and 6cm level, and the third test tube at the 3cm and 7cm level. Next you you start with test tube one and fill the first mark (1cm) up with catalase,the second (5cm) with H2O2, and last you stir, mix, wait 20 seconds, then record bubble height. With the second test tube you: fill the first mark (2cm) up with catalase, the second (6cm) with H202, and last you stir, mix, wait 20 seconds, then record bubble height. And with the last test tube you: fill the first mark (3cm) up with catalase, the second (7cm) with H202, and last you stir, mix, wait 20 seconds, then record bubble height. Exercise Four: Effect of ph on enzyme activity One may begin by gathering three test tubes, labeling them at the 1cm, 3cm, and 7cm levels, and then filling them each up to the 1cm mark with catalase. Next you start with filling test tube one with the ph3 solution up to the second mark. Then you proceed by filling the third mark up with H2O2, wait a minute, swirl, mix, wait 20 seconds, then measure and record bubble height. Second, you get test tube two and fill the second mark up with the ph7 solution. Then you proceed by filling the third mark up with H2O2, wait a minute, swirl, mix, wait 20 seconds, then measure and record bubble height. Last, you obtain test tube three and you fill the second mark up with the ph11soultion. Then you proceed by filling the third mark up with H2O2, wait a minute, swirl, mix, wait 20 seconds, then measure and record bubble height. Results: Exercise One: This first exercise was done to show the people conducting the experiment that catalase did truly break down hydrogen peroxide. Something that may be hypothesized about this exercise before doing it is that there most likely will be no reaction with test tubes two and three. As you may see on

table one and graph one, the hypothesis is correct. Test tube one contained the only reaction and as for test tubes two and three they remained unchanged. These results were obtained by having to control experiments (the water/hydrogen peroxide and the catalase/sucrose mixtures) and the dependent variable hydrogen peroxide mixed with the constant variable (catalase). Exercise Two: This exercise was done to prove the point on how temperature may increase the reaction rate but also may denature the enzyme. One may hypothesize that the reaction rate can only be at it's highest point at only one certain degree. This is proven true by table two graph two. One thing that stood out the most to me was that the bubble height was higher at the 4D/C mark rather than at the 50D/C mark. The first test tube was 80mm at 4D/C, the second test tube was 90mm at 37D/C, and the third test tube was 0mm at 50D/C. These results were acquired by putting three test tubes in three different temperatures with the same amount and kinds of substances with in them. Exercise Three: This exercise was done to prove the point that with the right amount of concentration the reaction can reach optimum speed, but with no enough, or too much concentration the reaction may react slower. So one may hypothesize that after a certain point of adding too much concentration the reaction rate will begin to slow down. Table three and graph three show that as the concentration increases the bubble height increases. So this draws the conclusion that there must be more room for additional concentration, because the bubble height only increased in this set of experiment. These results were obtained by having different amounts of catalase mixed with different amounts of H202. Exercise Four: This exercise was performed to show that too little ph will not get the fastest rate and too much wont either, but just enough will get the quickest reaction rate we are looking for. So from reading the information in the lab report one may hypothesize that the neutral ph7 may be the fastest reactant. And sure enough the results were obtained by mixing different ph levels with water and H202 to give us the conclusion that the ph7 was the fastest reactor. This is proved true through table four and graph four. Figure Legends: Table one: This table shows us the reaction between catalase and hydrogen peroxide. Table Two Graph One: This table and graph show an increase towards the beginning and then a decrease towards the end. Table Three: This showed a complete positive correlation, leading me to the conclusion that more concentration was available to make this reaction faster before slowing it's rate down. Table Four & Graph Two: This table and graph are just like graph one, it starts off with a positive correlation then ends with a decrease. Discussion: As proved by the results of the tables and the graphs you can see that not all the hypotheses were correct. For example, in exercise two one possibly would not have seen test tube one having such a high bubble height and test tube three having such a low bubble height. As for exercise three you could draw a hypothesis that the 2cm test tube would obtain the highest bubble height and the other two would be below it. But sure enough that is not how it turned out. The 1cm test tube was the lowest, the 2cm test tube had the middle height, and the 3cm tube had the highest height. And last as for exercise four, the ph11 contained a lot more bubble height than the lowest ph. One may have made a hypothesis that the lower and higher ph's would have been close in height, but they weren't, test tubes two and three were more closely related. I believe that my data may differ from others but I do not believe that it is way off, I

feel that it is fairly accurate. In conclusion, all enzymes are proteins that catalyze reactions by lowering their activation energies. So I believe it's safe to say one should include a decent amount of protein intake throughout the course of a day to improve metabolism.