Effects Of Temperature Ph And Gelatain On

Effects Of Temperature, Ph, And Gelatain On Bromelain Essay, Research Paper

Effects of Temperature, pH, and Gelatin on BromelainIntroduction

Bromelain is an organic catalyst that speeds up the rate of chemical reactions. Bromelain is used in this experiment to catalyze the break down of proteins and is called a proteolytic enzyme. Enzymes enable reactions to occur at a lower temperature and energy levels. This allows many reactions to occur inside living organisms that would otherwise have been impossible to take place. This enzyme is found in fruits, leaves, and stems of many species of pineapple. Pineapple is a tropical fruit native to northern South America. It is referred to as a multiple fruit because it consists of enlarged ovaries of many flowers more or less grown together into one mass. Ananas comosus is the type of pineapple used in this experiment. The bromelain is found in the pineapple attached itself to the substrate and broke it apart. The time required to complete the disassociation of the substrate was very important to the results of the lab, therefore time keeping must be precise in the experimentation. The effects of bromelain was compared under different temperatures and pH levels, the effects of bromelain were also observed in gelatin, there were three experiments performed and three hypothesis were formed. Temperature experiment: the higher the temperature the faster the reaction will occur to a point. The temperature will become to hot around 60 degrees Celsius and the enzyme will under go denaturation. Proteins are structurally complex and are very diverse. They have a high molecular weight and consist of amino acids liked by peptide bonds. Proteins are divided into three levels of structure: primary, secondary, and tertiary. Although some proteins have a fourth level which is the quaternary. Gelatin Experiment: The fresh pineapple juice will not solidify in gelatin. The hot juice and the control will solidify in the gelatin mixture.pH Experiment: The hypothesis for this experiment is that only a few numbers of pH will cause a reaction within a narrow range. pH levels at the extremes will cause the enzyme to under go denaturation. (Unfolding of the enzyme, loosing its ability to ac as an enzyme)Materials and Methods In the first experiment, effect of temperature on rate of enzyme activity, two test tubes were acquired for each temperature difference: 20 degrees C, 30 degrees C, 40 degrees C, 50 degrees C, and 70 degrees C. Each test tube was then cleaned with soap and water and rinsed out. The first five test tubes were then filled with five ml of fresh pineapple juice from the dispenser and labeled. The second five test tubes were then filled with five ml of a pH 3.5 buffer. The two test tubes labeled 70 degrees C and 70 degree C buffer and then placed in a test tube rack in an electronic water bath. Next, 20, 30, 40, and 50 degrees C water baths were made and the other four sets of test tubes were put into them. The bath temperatures were maintained throughout the experiment. All the test tubes sat in their respective water bath for five minutes to equilibrate. Next, filmstrips were placed in each of the test tubes and time began to be recorded. Every ten minutes the film was vigorously agitated for five seconds in the tubes in the 70 degrees C bath. In the 20 degrees C bath every five minutes the film was vigorously agitated for five seconds and checked for clearing. In the 30, 40, and 50 degrees C baths filmstrips were vigorously agitated every one minute and checked for clearing. When each of the test tubes cleared off their respective filmstrip then the time was recorded on the table as to how long it took. The fastest reaction happened in the 50 degrees C tube with a time of three minutes. Then the 40 degrees C tube was next to react at five minutes. The 30 degrees C tube was third to react at a time of 11 minutes. After twenty the test tube in the 20 degrees C reacted. The tube in the 70 degrees C never reacted. After the experiment was complete, the contents of the test tubes and strips of film were dumped into a bucket. The test tubes were then washed and dried out.

In the second experiment, effect of bromelain on gelation of gelatin, four clean test tubes were attained. The first test tube was labeled fresh juice and filled with one ml of fresh pineapple juice using a Pasteur pipette. The second test tube was labeled hot juice and then filled with one ml of the pineapple juice heated to 70 degrees C using a Pasteur pipette. The third test tube was labeled canned juice and then filled with one ml of canned pineapple juice using a Pasteur pipette. A fourth test tube was labeled control and filled with the pipette to one ml with water and dispensed into the test tube. Next, one ml of JELL-O was added to all four test tubes using a Pasteur pipette and the contents of the all four test tubes were mixed by placing a piece of wax paper over each tube and shaking it. Then all four test tubes were placed in a beaker of ice for five minutes. After five minutes in the ice, all four test tubes were removed and tilted slightly. The consistency of the gelatin in each test tube was observed and recorded in Table 2. The unused ice was then returned into the cooler. The test tubes were then emptied and washed out. The results of this experiment were that three test tubes turned to gel and one was still a liquid. The only one that didn t gel was t