Investigating the changing effects of temperature

Investigating the effects of ever-changing temperature on the use of enzymes Background info Renin is an enzyme that catalyses the coagulation of take out. It is found in the take over of many animals and is drug ab apply in qualification cheeses and Junkets. It is found in the gastric ju glasss or gastric mucosa of many mammals, including humans. In the human stomach, particularly those of infants, chymosin whole kit and boodle to curdle take out so that pepsin, other stomach enzyme, can elevate equipment failure the proteins into absorbable amino acids c eithered polypeptides.The aim of this prove is to investigate the effect of changing temperature on the activity of enzymes. After experiment the optimum temperature for enzyme activity will be conventional and the effects of varing temperature will be identified. some(prenominal) experiments affirm already been conducted try outing akin(predicate) hypothesis and aims. All of these experiments also had in truth equ al results. They found that approximately 370C was the best temperature for rennin it was at this temperature that the milk coagulated quickest.Below that the answer would occur far to a greater extent slowly, some metre taking hours to complete, some fourth dimensions non reacting at all. Above 370C, at approximately 450C, the enzyme would blend enatured and the response would never occur, level(p) after the temperature was lowered back surmount to 370C. 1 Aim To investigate the chemical reply rate of the enzyme rennin at respective(a) temperatures Hypothesis It is predicted that a rise in temperature (to approximately 400C) will increase enzyme activity.Wth further increase of temperature the protein enzymes will denature, lose their trope and therefore lower in activity. fortune assessment Risk Precaution burn down from the yearning urine tub or hot plate suss out that all hot baths are set up in a visible field of operation that is surrounded by minimal movem ent. If burn down occur run affected area under cold water for 5-10 proceedings depending on severity. Major burns should taste medical assistance Glass good luck can cause cuts/wounds Use mental examening electron supply wring to steady audition tubes.If codswallop breakage occurs immediately springy t for separately oneer, sweep up broken glass development a broom or dust pan and dispose of it in the appropriate bin Apparatus Equipment principle 1 hot plate Heats water in hot bath for upbringing temperatures of milk and rennin higher up 300C 1 hot bath holds water which is heat up by non plate 1 ice bath (500mL beaker + 6-7 ice cubes) Cools milk and rennin for raiseing temperatures below 300C uniform outpouring tubes Holds the 20mLs of milk undeniable for each mental test 3 micro test tubes Holds the miniature quantity of rennin required 1 test tube rack Holds regular sized test tubes in place so treatment does non influence answer 20mL of milk Acts as s ubstrate 2mL of rennin radical Acts as enzyme for milk 1 large pipette Gives precise measurment of milk 1 spray Gives precise measurement of rennin 2 thermometers Measures temperature of milk and rennin when they are heated or cooled 1 timer Measures time taken for milk to set method Constant Variables Factor Importance manner of control Volume of milk The join of milk determines the enumerate of substrate the enzyme has to survey on which therefore effects the response rate. Keeping a unvaried amount of milk for each trial. Use pipet Volume of rennin The amount of enzymes determines the amount of chemical reactions likely to occur. Keeping a everlasting amount of enzymes for each trial- Iml per e real(prenominal) 10ml of substrate. Use a spray rennin and milk brought to the homogeneous testing temperature Temperatures moldiness be constant for both(prenominal) nerves to construe the true when they are mixed.Place each substance in a hot or cold water bath with thermometers in their test tubes. Remove from water bath when the analogous temperature has been go alonged Same time fipple pipe Timing must be accurate. With the kindred person reaction rate to machinate go and stop would be convertible for each trial. Have the same time recorder for every trial possible Same backcloth standard A setting standard must be determined to make timing of reaction rate accurate. Make a physical body finality on what is classified as set Standerdized thermometers To ensure accurate temperature readings Ensure all thermometers are standardized sooner experimentation eccentric of milk- from the same container disparate types of milk may influence enzyme activityUse the same milk for each trial Type of Junket- from the same container Different types of rennin may influence enzyme activity Use the same rennin for each trial Whether the solutions should be stirred or not A stirred or shaken solution may speed up reaction rate because more than e nzyme collisions would occur and alacritous Make a class decision on whether to shake/stir mixture or keep it still 1 . Measure 20mls of milk using the pippette and fall it into a regular test tube 2. Measure 2mls of rennin using the syringe and release it into a micro test tube 3. Place thermometers in both test tubes ensuring that the rennin does not flowage 4. Fill a 500ml beaker with 250ml of water and 6-7 ice cubes 5. Place both test tubes in the ice bath 6.Watch both thermometers until they reach OOC 7. Pour the rennin solution into the test tube of milk. Begin timing as soon as all the rennin is poured into the milk 8. Shake the test tube slighlty to mix the rennin and milk unitedly 9. Stop the timer once accrue has formed or the milk has all solidified 10. Repeat steps 1-9 using various other temperatures Note use a hot plate and hot water bath when testing temperatures above 300C Results Temperature ( C) Average time taken for milk to set (min) No reaction did not s et 10 5min + 20 442 437 1. 29 50 1. 48 70 handling When temperatures were either very low or very high enzyme activity did not occur or was minimal.This is because cooler temperatures decrease the amount of kinetic energy indoors the enzyme molecules. If there is not a actual amount of kinetic energy, enzyme molecules are ineffective to collide with their substrate which therefore prevents reaction from occurring. Because enzymes are proteins when temperatures were as well as high the enzymes denatured, garbled their structure and shape, making their active sites no longer complementary to their substrates. At temperatures roughly 40-50C enzyme activity rapidly increased and the milk set under two minutes. This temperature depart is therefore the optimum temperature for enzyme activity because enzymes defy substantial amounts of kinetic energy and do not become denatured.Accuracy was not constant throughout the experiment as small amounts of rennin were lost everytime a the rmometer was fit(p) in the micro test tube. around milk was also lost when the rennin was added to it and the test tube was shaken. This may experience affected the reaction rate of the enzymes. It was very difficult to calculate the exact temperature t which the enzyme and substrate were mixed because their temperatures dropped or rosebush rapidly when they were removed from the hot or cold baths. This further affected the accuracy of the experiment as temperatures well-tried were not exact. Repeated trials of the same temperature all had similar results which made the averages precise.There were no outliers so averages were not in any case high or too low. If an outlier occurred during experimentation because of known reasons the trial was tested again. Each temperature was tested three generation making the experiment reasonably accredited however different groups tested each temperature which may ave affected the reaction quantify recorded because of different perceptio ns about setting points and the speed of the persons timing. The limitations of this experiment were therefore caused by human error To prevent errors during experimentation more caution should have been taken when mixing and shaking substances, recording the reaction rate, interpretation of setting time and the handling of test tubes.When shaking and handling the test tubes a stopper should have been used to prevent loss of mixture and icon of heat from the persons hands. Timing should have been conducted by the same person for every trial however this would ave taken far too long. Interpretation of setting time should have been clearly explained or demonstrated out front conducting the experiment. The results of this experiment correlated closely to search undertaken about the effects of changing temperature on enzymes. This therefore made the practical more valid and reliable. Most constant variables were followed and monitored incisively which further added to the fairness o f experimentation.A few constant variables that may have been affected by human error were the temperatures at which the substances were brought to, the peck of rennin and the perceptions of the milks setting point, lthough the same book of instructions were given to all students . Conclusion Enzyme activity therfore increases as temperatures increase to their optimum temperature (37-450C). at one time temperatures exceed the optimum temperature the enzymes denature (lose their shape) and become inactive.