The hypothesis, in relation to the enzymatic activity by rabbles such as the substrate concentrations, temperature, PH and chemical interactions on the rate Of reaction, stated the following scenarios: (1) If the substrate concentration is increased, then the enzymatic rate will increase (2) If the temperature is increased, then the enzymatic rate will also increase (3) If the PH level is increased, then the enzymatic rate will decrease.
To test the effect of a substrate concentration on enzyme activity, the amylase enzymes were combined with a different substrate concentration (starch) and the rate of the reaction was determined with the aid of kill. If starch was detected, the solution turned to dark blue; if the starch was already broken down, then reaction stayed colorless. To test the optimal PH. The starch and a buffer were combined at a specific PH level and the rate of reaction was tested.
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To determine the optimal temperature of amylase enzyme, the solution and amylase enzyme were held at various temperatures and the rate of reaction was determined. The rate of reaction for amylase enzyme with less of the substrate concentration (1 % starch) with concentration of O. 125% happened at a faster rate of only 20 seconds. The rate of reaction for amylase enzyme tit the effect of varied PH occurred at a quicker rate with PH range of 5-6 close to neutral PH of 7. 0 in less than 10 second.
Finally, the rate of reaction for amylase enzyme at different temperature happened faster at ICC (close to body temperature) as compared to other temperatures. The results from collected data showed that the smaller the concentration of starch in the amylase enzyme, the faster the rate of reaction and the quicker the breakdown of starch; note: the PH 7. 0 was the optimal PH for amylase activity, and the temperature of 37 co was the optimal temperature for amylase enzyme activity. Introduction: Enzymes are of primary importance in many chemical reactions which take place in living organisms.
When digestion occurs, enzymes released into the mouth, stomach, and intestines catalyst or accelerate reaction which results in the breakdown of large food molecules into small building block molecules. The molecules upon which an enzyme acts are called substrates. Each enzyme binds to the substrate; this binding process is called the “Lock and Key Theory” (I. E. The active site in the enzyme is the keyhole and has a very specific shape. This specific keyhole can only bind with a specific key which will match the keyhole. The key is the substrate molecule.
Once the key binds with the keyhole, the reaction takes place. ) This process significantly reduces the activation energy necessary for a bound substrate to undergo a particular reaction. Every enzyme has an optimum PH and temperature range at which the enzyme is structurally most stable and most active, and for which the us abstract most easily undergoes the binding process. Any environmental conditions which destroy protein molecules will also eliminate enzymatic activity. The rate of the reaction or enzyme activity can be changed.
The environmental factors influencing enzyme activity are the concentration of substrate (or enzymes themselves), the effect of PH, and the variance in temperature. The rate of an enzymatic reaction rises with increase Eng temperature because substrates collide with active sites more constantly when the molecules move quickly. However, the speed of the enzyme reaction falls severely above or below that temperature. The optimal temperature permits the utmost number of molecular collisions and the states alteration of the reactants to product molecules.
Most human enzymes have an optimal temperature of about 35-ICC (close to body temperature 37 ‘C). In addition to an optimal temperature, every enzyme also has optimal PH values at which it is the most active. The optimal PH value for most enzymes fall in the range of PH 6-8 (close to neutral PH 7); however some digestive enzymes in the human stomach work best at very acidic PH of 2. In this lab experiment the action of the enzyme Amylase was observed on starch (the substrate). Amylase changed the starch into a simpler form, the agar maltose, which is soluble in water.
Maltose then breaks down the glucose chains of starch in the pancreas and intestines. Amylase is present in human saliva, and begins to act on the starch in the food while still in the mouth. Exposure to heat or extreme PH (acid or base) will denature proteins. Enzymes, including amylase, are proteins; if denatured enzymes can no longer act as a catalyst for the reaction. In the presence of potassium iodide, starch turns a dark purple color; however maltose does not react with 121Enzymes have an ideal range of values for any of the variables, or optimal conditions, in this experiment. When these optimal conditions are exceeded or not met, the enzyme will be much less effective. Therefore, in this experiment the optimal condition of amylase was tested base on the three independent variables which affected amylase’s efficiency. The hypothesis concerning enzyme activity and reaction rate, as influenced by variables such as Substrate concentrations, temperature, and PR, include the following statements: ; If the substrate concentration is increased, then the enzymatic rate will increase. If the temperature is increased, then the enzymatic rate will also increase. ; Fifth PH level is increased, then the enzymatic rate will decrease. Materials and Methods: The general method that has been used to observe the enzymatic reaction Was the detection of starch in the substance. If there Was no starch remaining, it’s assumed that it has been completely hydrolysis and the amylase was successful. The reaction rate it took for that to occur was the efficiency of the enzyme, and it varied as the independent variable varied.
The dependent variable in all of the reactions was time, or the rate of hydrolysis. Therefore, it was important to method check, with the aid of a chemical compound (potassium iodide), each of the independent variables substrate concentration, temperature, and PH effect, for detection of starch. Procedure 1: (Effect of Substrate concentration) 1. Prepare the starch dilution set: Four standard test tubes labeled IA through AAA were obtained and 5-ml distilled water was added to each test tube. Then, in test tube IA 5-ml of starch was mixed in and approximately 5-ml of this solution was added to tube AAA.
In tube AAA, 5-ml of tube AAA was mixed and about S-ml of this elution was added to tube AAA. 2. Prepare the experimental test tubes set: Four standard test tubes labeled 1 B through B were obtained and about 2- ml of tube AAA was transferred into tube B. Same procedure was repeated for remaining tubes B through B from test tube AAA through IA; after these transfers, tubes IA-AAA were discarded. 3. Approximately 40 drops of the buffer solution (PH 7. 0) was mixed into each tubes 1 8-B and the test tubes were placed aside. 4.
Prepare the test plate: A test plate was obtained and 2 drops of kill were added to each compartment of the test plate with the plastic pipette. Beginning with test tube B, approximately I-ml Of amylase solution was mixed in the tube; immediately, the initial time zero was recorded and one drop of this mixture was immediately transferred via a disposable pipette into the compartment on the test plate. The reaction mixture was then sampled at 30 sec intervals until the potassium Iodide solution remained yellow-amber (indicating the digestion of all the starch) or until 10 minutes has elapsed.
This same procedure was repeated for remaining test tubes 38-B and all of the observations were recorded in the Data tables for analysis of the effect of abstract concentration. Procedure 2: (Effect of PH) 1. Prepare the test tubes Five standard test tubes were obtained; the tubes were numbered 3, 5, 6, 7, and 9. Approximately 5-ml of the appropriate buffer (3, 5, 6, 7, and 9) was added to each tube; then about 1. 5 ml of amylase solution was added to each tube. 2. Prepare the test plate: A test plate was obtained and 2 drops of 121Using the tube labeled 3: Approximately 2. 5 ml Of the 1 % starch solution was transferred into the elution tube and the initial time zero was immediately recorded. Then, one drop of this mixture with a disposable pipette was transferred into the compartment on the test plate. The reaction mixture was sampled at 30 sec intervals until the potassium Iodide solution remained yellow-amber (indicating the digestion of all the starch) or until 10 minutes has elapsed, and same procedure was repeated for remaining test tubes 5, 6, 7, and 9; all of the observations were recorded in the Data tables to indicate the effect of different PH levels.
Procedure 3: (Effect of temperature) 1. Prepare the sample test tube: Four standard test tubes were labeled IA through AAA, and approximately 2-ml of the 1% starch solution was transferred into each tube. This was then followed by 4-ml of denizen water and I-ml of the buffer solution (PH 7. 0) being added to each tube. After preparation of the solution in each tube, tube IA was held in room temperature (ICC), tube AAA was placed on ice beaker (ICC), tube AAA was placed in ICC water bath, and tube AAA was placed in ICC water bath for 10 minutes. 2.
Prepare the second set of test tubes: Four standard test tubes were labeled B through B and about I-ml of amylase was transferred into each tube. Tube B was then held at room temperature (ICC), tube B was placed on an ice beaker (ICC), tube B was placed in a 37 co water bath, and tube B was placed in a ICC water bath for 10 minutes. 3. Prepare the test plate: A test plate was obtained and 2 drops Of 121One drop of this mixture via a disposable pipette was then transferred into the compartment on the test plate. The reaction mixture was sampled at 30 sec intervals until the potassium Iodide solution remained yellow-amber (indicating the digestion of all the starch) or until 10 minutes has elapsed, and the same procedure was repeated for remaining test tubes B with AAA, B with AAA, and B with B; all of the observations were recorded in the Data tables to indicate the effect of different temperatures. Rest Its: 1.