An Investigation Into the Effects of Temperature on Enzyme Action

An investigation into the effects of temperature on enzyme action An enzyme is a biologic catalyst that speeds up the rate of reaction in certain biological give-up the ghosts. They play a vital role in many aspects of humankind physiology and are necessary for the functioning of a recite of systems, for typeface in the digestive system to help to break follow fall out food. either enzymes conduct a unique active site that can fit on to a particular molecular arrangement on a object lens substrate a substance e. g. carbohydrate, protein, or fat, that the enzyme is designed to breakdown.There are a number of different enzymes in the human body from each one attri providede produced specifically to perform a certain role. Enzymes are non themselves destruct in the reaction to break down a substrate but their effectiveness is reduced under certain adverse environmental conditions. The devil most important ones are temperature and PH level also immersion of enzyme is also a determining factor. Enzyme action is biochemical in spirit and, in line with many chemical reactions it speeds up with an increase in temperature. This would continue until a certain critical temperature is reached where its running(a) faculty drops suddenly.This is due to a deforming or de-naturing of the active site leaving the enzyme unable(p) to bond with its substrate. In humans this can shit life hard consequences if the body temperature rises above 40 degrees Celsius. As enzymes normally function in their own particular part of the anatomy, they operate most efficiently in a medium with a specific acid/alkali (PH) rest period. If this balance is either too high or too low its functioning is reduced and in extreme circumstances this can last to de-naturing also. An investigation was carried out in order to study the functioning of a particular enzyme Trypsin.This is produced in the pancreas and is found in the pancreatic juice it is utilize to break down proteins. As it is believed that all enzymes function most efficiently at some optimum (ideal) temperature this investigation looked at the ability of trypsin to function effectively over a range of discreet temperature settings. These included touchstones from 20 degrees c, to 60 degrees c, in ten degree intervals and included a measurement at 0 degrees c, for comparison. The substrate use was protein from skimmed milk. Method A one per-cent solution of trypsin in sodium hydrogen carbonate was used to eliver a uniform dose in a favourable P. H. environment. A skimmed milk powder preparation of 5% in 95% purified water was used to mitigate undue discrepancy in the moderates. any equipment, that came into contact with the study mediums, was sterile e. g. trial tubes, syringes etc. A gull was marked with a marker pen, on the side of one test tube at the bottom and because 2ml of trypsin solution was then primed(p) in a second. The two test tubes were then placed in one of a number of wa ter baths, set at vary temperatures, for cardinal minutes, in order to raise them (or subvert) to a target temperature.The period was measured with a digital stop clock. After five minutes both samples were removed and the trypsin was poured into the tube containing the milk. The stop clock was re-started and ran until the trypsin had broken down enough of the milk protein to make the marker pen cross understandably visible across the width of the test tube. This process was repeated, by five experiment groups, until each group had a measurement at each required temperature. The results were then recorded on a data table. All due health & safety precautions were followed including the wearing of lab coats and goggles.Care was interpreted when discourse the samples at the upper temperature ranges e. g. 60 Celsius. As trypsin itself is an irritant care was taken with this and hands were washed thoroughly. Nobody drank the milk. One or two checks and tests were localise in plac e in order to validate the data trends. A colorimeter was used as a more scientifically reproducible measure of milk clarity (one sample was taken from each temperature setting) and crocked readings were calculated from the table results to hopefully reduce the effect of moderate outliers.Conclusion As expected the results did indeed show an increase in trypsin action as the temperature was raised above zero towards forty degrees Celsius. The only(prenominal) surprising result was that the data trends showed that this increase was sustained until a maximum was reached at 50 degrees c, when it was expected that the maximum would be at 40 degrees c, which is closest to its working temperature of around 37 degrees c. There are a couple of achievable explanations for this apparent anomaly.Firstly Among the readings for the 40 degrees c, and 50 degrees c, samples there are a number of irregularities including two low readings at 50 degrees c. It must be spoted out at this time that a shorter time recording indicates an increase in trypsin activity, so the table and graph are ones of inverse correlation as the graph or data go down, the value (enzyme activity) goes up. Secondly The samples were only placed in the water bath for five minutes and as soon as any samples were removed the temperature would have begun to gravitate towards board temperature.It stands to reason that the rate of change in samples that were furthest from room temperature, to start, would have been the greatest. Consequently, the sample that should have been 50 degrees c, to start, might have been considerably lower at the time of reaction. It is hard to believe that all of the trypsin would have been de-natured in one instant, so any that was not would have reacted rapidly make a fast reaction time for the 50 degrees c, sample.Returning to the first point As the trypsin reacted during the experiment, the marker pen cross would have appeared gradually as the milk sample cleared. This w ould have left a large tolerance for error because the point at which to stop the timer would have been, rattling much, a matter of opinion. This has probably lead to a number of inconsistencies in the results. The effects on the data range seem to have been varied, for example At 0 degrees Celsius, the difference between the highest and lowest recordings was 6%, at 60 degrees it was 241%.One check, previously mentioned, was the measurement of a data set use a colorimeter. This device measures the amount of light passing through a sample. The results from the back-up experiment confirmed the expected trends namely as the temperature increased the trypsin activity increased up to an optimum of 40 degrees c, after which the enzyme activity tail off rapidly. How the experiment could have been improved Overall, the experiment was well conducted and approximate practice was followed.However, having one person with experience in analyzing samples deciding precisely when to stop the timers, would have reduced the occurrence of any outliers in the data. Also, it is not difficult to imagine setting up equipment that would work along the lines of the colorimeter, step light levels through a sample, only set to stop a timer the moment a particular uniform reading was reached. The samples could then be placed in this equipment the moment the trypsin was added, hopefully resulting in an surgical and uniformly consistent set of data being extracted.