Part C: Change the amount of the substrate First, the blank was prepared according to table 2 without the enzyme addition. The enzyme was added later after the blank was measured by the spectrophotometer. Table 2: The amount of Sodium Phosphate Buffer pH 7.0, L-Dopa, and enzyme needed in each cuvette. Cuvette 1 Cuvette 2 Cuvette 3 Cuvette 4 Cuvette 5 Sodium Phosphate Buffer PH 7.0 (mL) 2.40 2.20 1.80 1.60 1.10 L- Dopa (mL) 0.20 0.40 0.80 1.00 1.50 Enzyme (mL) 0.40 0.40 0.40 0.40 0.40 For example, to prepare the cuvette 1, 2.40 mL of buffer pH 7.0 was measured by the micropipette P-1000, and was added into cuvette labeled #1 for the second set of cuvette. Next, 0.20 mL of L-dopa was measured by the micropipette P-1000 and was added into …show more content…
Then, the cuvette that labeled #1 was wiped off with the KimWipe and placed in the single cuvette holder in position 1 in the sample compartment. The position 1 was making sure aligned with the light source. The sample compartment door was closed, and was pressed “auto zero” button on the keypad. Then, after 30 seconds, the absorbance that displayed on the screen was read again and it was 0. The absorbance was read at 0 seconds, at 30 seconds, at 60 seconds, at 90 seconds and at 120 seconds. All the absorbances were remained 0 for the blank. After 120 seconds, the blank was then removed, and the appropriate amount of enzyme Tyrosinase (0.40 mL) was measured and added into the blank (cuvette #1) using the micropipette P-1000 according to the table 2. The final volume in the cuvette was 3mL. The cuvette contained the enzyme sample was wiped off with a KimWipe and was placed into the sample compartment of the machine. The sample compartment door was closed. The absorbance that displayed on the screen was read and documented in table 5. Then, after 30 seconds, the absorbance was read again and documented in table 5. The absorbances were read at 0s, at 30s, at 60s, at 90s, and at 120 seconds. Cuvette 2, 3, 4, 5 were followed the same method as cuvette 1 but different amount of buffer, L-dopa according to table 2. All the absorbances were read and documented in table
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The cuvette was placed in the spectrophotometer with the arrows, on both the cuvette and the SpectroVis, facing the same side. After the recording, the cuvette was removed from the SpectroVis and the content was poured back into the original volumetric flask. The absorbance as well as the maximum wavelength of each solution was recorded in Table 3 and
LABORATORY REPORT Activity: Enzyme Activity Name: Natalie Banc Instructor: Elizabeth Kraske Date: 09.22.2016 Predictions 1. Sucrase will have the greatest activity at pH 6 2. Sucrase will have the greatest activity at 50 °C (122 °F) 3. Sucrase activity increases with increasing sucrose concentration Materials and Methods Effect of pH on Enzyme Activity 1. Dependent Variable amount of product (glucose and fructose) produced 2.
Starch solution is then placed into the test tube at a quantity of 5 mL. 5 drops of Lugol’s Iodine solution is added to the test tube. If the color changes, then it is known that starches are present in the solution. Proteins are next tested. In order to do this, 5 mL of gelatin solution is added to the test tube. 10 drops of Biuret’s reagent are added to test for protein.
1 “substrate” and another “ enzyme.” Instead of using the distilled water, this time you are going to use different pH buffer in the enzyme test tube. In the substrate tube, add 7 mL of distilled water, 0.3 mL of hydrogen peroxide, and 0.2 mL of guaiacol for a total volume of 7.5 mL. For the enzyme tube, instead of distilled water add the pH solution (3) and 1.5 mL of peroxidase which equals a total volume of 7.5 mL. Use the dH2O syringe for our pH solution. To clean the syringe, flush it by drawing 6 mL of distilled water.
Record the amount of absorbance by converting transmittance every 5 minutes for a total of 20 minutes. Repeat all of these steps for the cantaloupe, banana, replacing the blank each time to recalibrate the spectrophotometer. After recording all the percent transmittance value, the data was then converted into absorbance value by using the absorbance conversion table. The information was placed on a plotted graph
5 water bath were set up each to10 °C. (5 were used do the experiment faster) 5 cm3 of starch solution were added into the 5 test tubes that were labeled test tubes. Then 5 cm3 of amylase enzyme was added into the other 5 test tubes that were labeled. Put one of the starch solution test tube (preferably the one labeled 1) and one of the test tube containing amylase into the water bath (10 °C).
Along with being found in plants, they are also present in liver cells, kidney cells, leukocytes and erythrocytes. For the concentration of enzyme experiment, the hypothesis was if the concentration of an enzyme increases, then the enzyme activity will increase as well. The hypothesis was proven to be true, because there are more enzymes to react with substrates. For the enzyme—factors affecting, the hypothesis concluded was if the temperature increases, than the enzyme activity will increase. This however was proven wrong, because enzymes become unstable at higher temperatures.
By observing figure 3, the more enzyme that is available, the faster the reaction rate is. The optimal enzyme concentration was chosen based on the R2 values from figure 2. The highest observable rate also had the best R2 number, which was closest to one. This enzyme concentration was used in part 2.
However, any doubts regarding the results may be traced to a few elements of the experiment that lend themselves to possible error. The following factors may have contributed to potential errors in the experiment; the need to zero the machine between each of the readings in obtaining the absorption spectrum and the resulting peak wavelength, the precision with which a person can accurately adjust the needle on the spectrophotometer to zero is limited, not putting in the inaccurate amount of cobalt chloride or water into the substance, and getting oil from our fingers onto the
Introduction Buffer is a solution that resists a change in pH when bases or acid are added. Solutions that are acidic contain high concentrations of hydrogen ions (H+) and have pH values less than seven. Buffer usually consist of a weak acid, and its conjugate base or a weak base and its conjugate acid. The function of buffer is to resist the changes in hydrogen ion concentration as a result of internal and environmental factor. This buffer experiment is important so that we relies the important of buffer in our life.
Catalase Activity on Substrate Based On Gas Pressure Production Rate Name of the Class Author’s Name Date Enzymes are organic compounds which act as catalysts and speed up biological reactions in biological organisms. They are not destroyed or changed during the reaction but rather they are used over and over again to catalyze many more reactions. Their activity may be affected and altered by factors such as temperature, substrate concentration, enzyme concentration and Ph.
Use these results to determine the product concentration, using Beer-Lambert’s Law: A= ɛCl (where A is the absorbance, ɛ is the molar absorptivity, C is the product concentration and l is the length of solution that the light passes through). Calculate the product concentrations at every minute for 10 minutes for all 7 of the test tubes using Beer-Lambert’s Law. Plot a graph of product concentration vs. time and then use the gradients of the 7 test tubes to determine the velocities of the reaction. After calculating the velocities, plot a Michaelis-Menten graph of velocity vs. substrate concentration.
Bio Chem lab Report 04 Enzyme Biochemistry Group Member: Chan Man Jeun Duncan (16002621) Law Sze Man (16000478) Introduction Enzyme is a protein base structure substance in our body. It works at a biocatalyst that will catalyzing the chemical reaction, which helps to speed up the chemical reaction. Enzyme could only function in specific shape, and the shape of enzyme is depending on the environment, therefore it is hard for an enzyme to function well in an extreme environment. The aim of this experiment is to see can the enzyme functions normally in different environment(pH, temperature and salt concentration) via using starch solution, amylase from saliva, 0.5M HCl solution, 0.5M NaOH solution and NaCl solution, and using iodine solution