Fungal And Bacterial Amylase During Starch Breakdown Biology Essay

fungal And Bacterial Amylase During Starch Breakdown biological science EssayThe lab conducted focused on examining the effects of temperature on the ability of fungal and bacterial amylase to segmentation amylum to maltose, and determine the temperature at which these two amylases work best, which is known as optimum temperature.The experimental part of the lab consisted in cathode-ray oscilloscope up the utensils that were going to be used during the actual experiment. During this section test tubes were labeled, and spot cuticles were outrankd in temperature/ m table created. For the second section of the experiment, tincture of iodine was placed in apiece row of the spot plates for each temperatures, and the solutions in the test tubes( bacterial , fungal amylase and amylum mixture) were tot uped to those same spots were iodine was added, depending on the time and the temperature corresponding to each amylase. The best temperature was deducted by observing the color change in the spot plates and comparing them with a color-coding scheme for stiffen hydrolysis.Conclusions for this undertaking were reached by analyzing the info collected by each group, which suggests that a change in temperature disturbs the activity of enzyme amylase. When exposed to low and high temperatures, these enzymes were not able to usage properly, therefore, reducing or eliminating their ability to breakdown certain compounds, especially starch. Enzymes need maintain at a certain temperature to be able to function at its optimal.IntroductionEnzymes be complex proteins produced by all living organisms with the function of enhancing chemical reactions through a process known as catalysis. During this process, the substrates, which are the molecules that result undergo the reaction, curbs to the active site of the enzyme to form different molecules called products. Each active site on the enzyme is unique, yieldting only substrates that match the shape of the acti ve site to bind to the enzyme in a process known as lock and key model, however, active sites are able to adjust their shape to permit the binding with a substrate through the induced fit model, which moves entire protein domains (Raven et al., 2008 Ringe Petsko, 2008 Whitehurst Van Oort, 2009).Catalysts, like enzymes, work by reducing the amount of energy required for a chemical reaction to take place by linking two substrates in the correct orientation or by accentuating chemical bonds of a substrate, which reduces the energy difference betwixt reactants and transition state. Enzymes are not consumed or changed during the reaction and they do not alter the equilibrium of the reactions they catalyze (Garcia et al, 2004 Raven et al., 2008 Whitehurst Van Oort, 2009 Alberte et al., 2012).The activity of enzymes is affected by multiple factors, including 1) pH (ranges from pH 6 to 8), 2) temperature (Rate of reaction increases with temperature, but only up to a point called optimum temperature. A change in temperature, e very below or above the optimum, causes the active site to denature, decreasing or preventing substrate binding. When exposed to low temperatures enzymes are not flexible enough to permit induce fit, and in high temperatures enzymes are too weak to maintain their shape.), 3) substrate concentration (If amount of enzyme is preserved constant and substrate concentration is little by little increased, the reaction velocity will increase until it reaches a maximum. After this point increasing substrate concentration will not increase the rate of reaction), 4) allosteric inhibitors and activators (Inhibitors are substances that bind to an enzyme and decreases its activity, and they can occur in two ways competitive inhibitors and noncompetitive inhibitors. Effectors that enhance enzyme activity are referred to as allosteric activators, which bid to allosteric sites to keep an enzyme in its active configuration), and 4) cofactors (Many enzymes req uired the presence of other compounds, called cofactors, which during the catalytic activity, A cofactor can be a coenzyme, a prosthetic group or a alloy ion activator (Harisha, 2006 Raven et al., 2008 Whitehurst Van Oort, 2009).Enzymes have a wide spectrum of functions in the bodies of living organisms they are present from signal transduction to generation of muscle contraction. The also break starch molecules, forming smaller fragments of maltose, which can be easily absorbed by mammals. And it is the ability of enzymes to breakdown starch and the effect of temperature during this process that will be analyzed in the lab (Whitehurst Van Oort, 2009 Alberte et al., 2012), expecting that the results collected confirm that temperature does have an effect in bacterial and fungal amylase activity.MethodsThe experiment should be performed once per group, using fungal (Apergillus oryzae) and bacterial amylase. Starch catalysis will be monitored by using iodine test, which turns from white-livered to rich- mordant in the presence of starch.Experimental SetupPlace a paper under the spot plates and label the top side with temperature values 0,40,60,95 C, and the side with the times 0,2,4,6,8,10 min. Obtain 4 test tubes and label each with a different temperature, enzyme source, either bacterial or fungal and group number. Repeat previous step, but this time include the letter S, which stands for Starch solution. Finally add 5ml of 1.5% starch solution into each of the test tubes labeled S.Effect of temperature in amylase activityAdd 1ml of amylase into each of the test tubes that do not contain starch, and place the 8 test tubes (4 containing starch and 4 containing amylase) into their respective temperatures, allowing all test tubes to equilibrate for 5 minutes. Add 2-3 drops of iodine to the first row of the spot plate corresponding to o minutes. After 5 minutes has passed and test tubes are equilibrated, head a few drops of starch solution from each temperat ure to the row where you added the iodine. teem the starch solution into the tube containing amylase without taking it put of bath, and set the timer for two minutes.Add 2-3 drops of iodine to the second row, and after 2 minutes has passes, transfer a few drops of the starch-amylase mixture from each tube to the 2 minutes row using the pipette correspondent to each temperature. After each special 2 min, add 2-3 drops of iodine and a few drops from starch amylase mixture. At the end of 10 min, note the temperature and the time at which 100% hydrolysis occurred. Repeat the act using the other amylase type, and using the color-coding scheme convert results into numerical values.Results temp (C)0406095Time (min)0555524.3333333.1666673544.1666673.0833332.83333356432.7558432.6833335103.83333332.755 put over 1 Class bonnie for Bacterial Amylase activityAfter all groups performed the experiment, a class data for bacterial amylase was collected. The average of the data was calculate and presented in give in 1, viewing color changes for each temperature.Temp (C)0406095Time (min)0555523.3333332.6666673.166667543.3333332.6666673.083333563.3333332.6666672.833333583.3333332.4166672.8333335103.3333332.4166672.8333335Table 2 Class average out data for fungous Amylase activityAfter all groups performed the experiment, a class data for fungal amylase was collected. The average of the data was calculated and presented in Table 2, showing color changes for each temperature. represent 1 Class Average for Bacterial Amylase activity Graphical RepresentationResults from Table 1 exposed in a graph, showing that all groups optimal temperature for Bacterial amylase is 60CGraph 2 Class Average Data for Fungal Amylase activity Graphical RepresentationResults from Table 2 were exposed in a graph, showing that all groups optimal temperature for Bacterial is 40C enroll 1 twine coding-scheme for starch breakdownStarch hydrolysis color coding scheme is used to determine the optimal te mperature for each amylase during starch breakdown signifier 2 Bacterial amylase activity spot plateGroup number 1 spot plate during bacterial amylase experiment showing the amylase reaction during each temperatureFigure 3 Fungal amylase activity spot plateGroup number 1 spot plate for fungal amylase experiment showing starch breakdown during each temperatureGraph 3 Bacterial Amylase Activity graphical representationBacterial amylase activity data taken from table 1 showing that optimal temperature for this kind of amylase according to group 1 is 60CTemp (C)0406095ColorColorColorColorTime (min)0blue/ disgraceful5blue/black5blue/black5blue/black2blue/black4med dark-brown3.5 arc brown3blue/black4blue/black4light brown3light brown3blue/black6med brown3.5light brown3dark yellow2.5blue/black8med brown3.5light brown3med yellow2blue/black10med brown3.5dark yellow2.5med yellow2blue/blackTable 3 Bacterial Amylase activityGroup 1 recorded color changes for each temperature during breakdown o f starch by bacterial amylase, and it was represented in numerical values by using color coding scheme presented in Figure 1Graph 4 Fungal Amylase Activity graphical representationFungal amylase activity data taken from Table 4 showing that optimal temperature for this kind of amylase according to group 1 is 40 CTemp (C)0406095ColorColorColorColorTime (min)0blue/black5blue/black5blue/black5blue/black2light brown3dark yellow2.5light brown3blue/black4light brown3dark yellow2.5light brown3blue/black6light brown3dark yellow2.5light brown3blue/black8light brown3med yellow2light brown3blue/black10light brown3med yellow2light brown3blue/blackTable 4 Fungal Amylase ActivityGroup 1 recorded color changes for each temperature during breakdown of starch by fungal amylase, and it was represented in numerical values by using color coding scheme presented in Figure 1Temp (C)0406095Time (min)0000020.4082480.2581990040.2581990.2581990.258199060.3162280.3162280.418330080.3162280.3162280.3763860100.5 163980.3162280.4183300Table 5 Class Average Standard Deviation for Bacterial Amylase activityFrom the results from Table 1, the standard deviation was taken, showing that the results collected by each group for Bacterial amylase are penny-pinching to average results.Graph 5 Class Average Standard Deviation for Bacterial Amylase activity Graphical RepresentationData from Table 5 was exposed in a graph, showing that the difference mingled with the mean and the samples collected by each group is minimal.Temp (C)0406095Time (min)0000020.4082480.5163980.68313040.4082480.4082480.66458060.4082480.4082480.68313080.4082480.4915960.683130100.4082480.4498680.683130Table 6 Class Average Standard deviation for Fungal Amylase ActivityFrom the results from Table 2, the standard deviation was taken, showing that the results collected by each group for Bacterial amylase are close to average results.Graph 6 Class Average Standard Deviation graphical RepresentationData from Table 6 was exposed in a graph, showing that the difference between the mean and the samples collected by each group is minimalDiscussionAfter evaluating the results of the experiment, present in Table 1 and 2 it can be concluded that the data provides enough evidence to support the predictions or hypothesis presented in the introduction section that when temperature is not optimal for an enzyme, it will denature or reduce its functions. The results showed that low or high temperatures have an effect in the ability of enzymes to break down starch (Graph 1 and 2). By comparing the results with color coding scheme provided (Figure 1), the optimal temperatures for both amylases were able to be determined. The optimal temperature for the enzyme had a bright yellow color, which meant that the amylase was able to breakdown the starch present in the solution when the solution remained blue-black the enzyme is said to be denature, meaning that it was not capable of breaking down the starch( Figure 2 and 3).The most important parameters taken into account to get the previous results were temperature and time. Looking at the color for the reaction between starch and amylase, by using the Iodine test, it can be concluded that for bacterial amylase, the optimal temperature is 40 C, and this occurs around the 6 minute time. Fungal amylase optimum temperature was reached at 6 minutes time and it was 60 C. All the previous result can be observed in Figure 2 and 3, as well as in Graph 1 to 5Table 5 and 6 show that the results of the experiment are consistent for all lab groups, because the difference between the sample data collected by each individual group and the average of that data is minimal, showing that, the results collected by each group are close very close to be accurate.What parameters of the experimental design were important in the expected (or unexpected) results?The expectations for the experiment concurred with the results, because a previous understanding of enzymes was given in th e lab manual, however, the optimal temperatures were not exactly known because each enzyme works best depending on its environment. For future research, the range in temperature should be more variable, not only including official values, but negative ones. Also, if enzymes sources had more variation, it will provide a better understanding of the optimal conditions and temperature of enzymes.Literature Cited/ References Alberte J., Pitzer T., Calero K. (2012).General Biology Lab Manual / Second Edition. Florida world(prenominal) University The McGraw Hill Companies. Garcia-Viloca M., Gao J., Karplus M. Truhlar D. G.(2004). How Enzymes Work Analysis by Modern Rate Theory and Computer stimulations. Science 303pp. 186-195. Harisha S. (2006). Introduction to Practical Biotechnology. India Laxmi Publications. Raven P., Johnson G. B., Mason K. A., Losos J. B., Singer S. S. (2008). Biology 8th edition. New York The McGraw Hill Companies. Ringe D., Petsko G. A. (2008). How Enzymes Work. S cience 320 pp. 1428. Whitehurst R. J., Van Oort M. (2009). Enzymes in Food Technology Wiley-Blackwell 2nd edition.