LABORATORY REPORT
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Activity: Enzyme Activity
Predictions
1. Sucrase will have the greatest activity at pH 6
2. Sucrase will have the greatest activity at
60 °C (140 °F)
3. Sucrase activity decreases with increasing sucrose concentration.
Materials and Methods
Effect of pH on Enzyme Activity.
1. Dependent Variable. amount of product (glucose and fructose) produced
2. Independent Variable. pH 3. Controlled Variables. temperature; amount of substrate (sucrose) present; sucrase + sucrose incubation time
Effect of Temperature on Enzyme Activity.
1. Dependent Variable. amount of product (glucose and fructose) produced
2. Independent Variable.
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The human body would not be able to sustain a temperature of 104 degrees F for any length of time.
3. Specifically state where in the intestine sucrase is likely to be most active (pH along GI tract).
Small intestine (duodenum) is where the sucrase is likely to be most active.
4. Explain how sucrase activity in the intestine is affected by the sucrose content of food. Why is this important?
Sucrase activity in the intestine is affected by sucrose content in food as most foods contain some sucrose. Adding sucrose will increase sucrase activity.
5. Heat treatment was used to denature the sucrase that was added to the control test tube. In the experimental test tube, alkaline
DNS was added to denature sucrase to stop the enzymatic reaction. Discuss what temperature was probably used to denature sucrase and what the probable final pH was of each solution after DNS was added.
Heat treatment was most likely used as temperatures too high would cause the enzymatic reaction to decrease. The probable temperature that was used would most likely be 70 degrees C.
6. Restate your predictions that were correct and give the data from your experiment that supports them. Restate your predictions that were not correct and correct them, giving the data from your experiment that supports the corrections.
I predicted that sucrase will have the greatest activity at pH 6 based on Table 6 of data
In this lab or experiment, the aim was to determine the following factors of enzymes: (1) the effects of enzymes concentration the catalytic rate or the rate of the reaction, (2) the effects of pH on a particular enzyme, an enzyme known and referred throughout this experiment as ALP (alkaline phosphate enzyme) and lastly (3) the effects of various temperatures on the reaction or catalytic rate. Throughout the experiment 8 separate cuvettes and tubes are mixed with various solutions (labeled as tables 1,3 & 4 in the apparatus/materials sections of the lab) and tested for the effects of the factors mentioned above (concentration, pH and temperature). The tubes labeled 1-4 are tested for pH with pH paper and by spectrophotometer, cuvettes 1a-4a was tested for concentration and cuvettes labeled 1b-4b was tested for temperature in four different atmospheric conditions (4ºC, 23ºC, 32ºC and 60ºC) to see how the enzyme solution was affected by the various conditions. After carrying out the procedures the results showed that the experiment followed the theory for the most part, which is that all the factors work best at its optimum level. So, the optimum pH that the enzymes reacted at was a pH of 7 (neutral), the optimum temperature that the reactions occurs with the enzymes is a temperature of 4ºC or
Sulfuric acid - was used to stop the reaction with catalase and hydrogen peroxide. It denatured the enzyme (catalase) and halted the reaction so the amount of hydrogen peroxide decomposed could be measured.
• Fourthly, we kept the temperature at a constant 25°C using a water bath. At low temperatures, an increase in temperature causes an exponential increase in enzyme activity. This is because an increase in temperature provides more kinetic energy for the collisions of enzymes and substrates, so
Given the background, we hypothesized that for the first experiment, the lactase will break down lactose in milk and have a similar effect to sucrose. We also predicted that the Milk + Lactase reactant would have more glucose, the Milk + Water reactant would have a little bit of glucose broken down, the Sucrose + Lactase reactant would have less glucose than the Milk + Water reactant, and the Sucrose + Water reactant would have little to no glucose at all. As for the first procedure of the second experiment, we had hypothesized the more basic the solution would become, then the more glucose there would be. Our prediction for the first procedure of the second experiment was that there would be no glucose found in the solutions containing pH 4 and pH 7. For the second procedure of the second experiment, our hypothesis was that glucose would be present in the reactants at 4ºC and 25ºC while the reactant that had been in the hot water bath at 100ºC would have little to no glucose because it would have evaporated. We predicted that for this temperature experiment, the glucose would evaporate at 100ºC and would remain at 4ºC and 25ºC. For the first experiment we had found that a reactant of Milk + Lactase have high levels of glucose, while the other three reactants do not. As for the second experiment, for the first procedure, amounts of glucose were found in
Enzymes react differently under different conditions and concentrations, being the most productive at the enzymes specific optimum condition and concentration. The enzyme sucrase, extracted from yeast, breaks down the complex sugar sucrose into the simple sugar glucose. Testing for sucrase’s optimum environment, multiple reactions were ran using varying amounts and concentrations of sucrose and sucrase at different pHs and temperatures. The product was then treated with Benedicts solution to visually observe what amount of glucose was present after the reaction was ran; negative results being little to no glucose present and positive results being
1. Sucrase will have the greatest activity at pH 6 2. Sucrase will have the greatest activity at 40 °C (104 °F) 3. Sucrase activity increases with increasing sucrose concentration until a plateau is reached.
If temperature of the water(enzyme environment) is increased to 35°C, then the enzyme activity will
There is another digestive enzyme (other than salivary amylase) that is secreted by the salivary glands. Research to determine what this enzyme is called. What substrate does it act on? Where in the body does it become activated, and why?
amylase enzyme and the optimal temperature for fungal and bacterial amylase. In order to make
In part II of the lab six small glass tubes were obtained in a test tube rack. Ten drops of distilled water were then added to test tube 1, five drops to tubes 2-4, and no drops in tubes 5 and 6. Five drops of 0.1M HCl were added to test tube 5 and five drops of 0.1M NaOH to test tube 6. Five drops of enzyme were then added to all tubes except tube 1. Tube 3 was then placed in the ice bucket and tube 4 was placed in the hot bucket at 80-900C for five minutes, the remaining tubes were left in the test tube rack. After the five minutes five drops of 1% starch was added to every tube and left to sit for ten minutes. After ten minutes five drops of DNSA were then added to all the tubes. All the tubes were then taken and placed in the
During these experimental procedures, the implication of multiple different temperatures on fungal and bacterial amylase was studied. In order to conduct this experiment, there were four different temperatures used. The four temperatures used were the following: 0 degrees Celsius, 25 degrees Celsius, 55 degrees Celsius, and 80 degrees Celsius - Each temperature for one fungal and one bacterial amylase. Drops of iodine were then placed in order to measure the effectiveness of the enzyme. This method is produced as the starch test. The enzyme was tested over the course of ten minutes to determine if starch hydrolysis stemmed. An effective enzyme would indicate a color variation between blue/black to a more yellowish color towards the end of the time intervals, whereas a not so effective enzyme would produce little to no change in color variation. According to the experiment, both the fungal amylase and bacterial amylase exhibited a optimal temperature. This was discovered by observing during which temperature and time period produced a yellow-like color the quickest. Amylase shared a similar optimal temperature of 55 degrees Celsius. Most of the amylases underwent changes at different points, but some enzymes displayed no effectiveness at all. Both amylases displayed this inactivity at 0 degrees Celsius. At 80 Celsius both the enzymes became denatured due to the high temperatures. In culmination, both fungal and bacterial amylase presented a array of change during it’s
We did this to test which test tube would contain the highest concentration of glucose. What we wanted to know was if the lactase would affect the function of the enzyme. Our hypothesis is the lactase functions within a narrow pH and that will change in pH would affect the function of the enzyme. We predict that if we change the environmental factors it will have an effect the function of the enzyme if the pH is outside the range in its optimum activity. Our hypothesis was then proven because the reaction only occurred in a neutral and acidic state of pH, not basic. Which means the enzymes prime ability to function is a neutral, and acidic pH range.
Temperature has a negative and positive effect on enzymes. As the temperature increases from 0 to 40 degrees (See Fig 2) the movement of the enzyme and substrate quicken and will bind more often.But, as the temperature increase from 40 degrees the enzyme and substrate slow and cannot bind as quick and therefore at 63 degrees production stops.
At the temperature of thirty-seven degrees Celsius we found that the reaction of the solution would produce carbon dioxide. When the temperature decreased the activity of the enzymes dramatically slowed down, in resulting in less carbon dioxide
The major portion of an oral dose of sucralose (85%) is unabsorbed and is excreted unchanged in the feces of rats, mice, rabbits, dogs and man.. Sucralose is absorbed from the upper part of the gastrointestinal tract by passive diffusion (Grice & Goldsmith ,2000) . Of the small amount of sucralose that is absorbed following consumption (approximately 15% of an oral intake), most is also excreted unchanged. About 2–3% of an oral intake undergoes common phase II metabolism, specifically, glucuronidation . Both unchanged sucralose and its glucuronide conjugates are excreted in urine, and readily eliminated with no bioaccumulation(Grotz & Munro