3. Plasmid pBR607 is a 2.6 kb plasmid containing Ampicillin and Tetracycline resistance markers, an origin of replication, and unique restriction sites for the restriction enzymes EcoRI, BamHI, and PstI. Given the restriction map for pBR607 for the enzymes EcoRI, BamHI, and PstI, show on the agarose gel picture below where the approximate positions of the restriction fragments generated from the given restriction digests would be located after carrying out electrophoresis. 4. As part of an undergraduate project, a student was attempting to construct a restriction map for the plasmid pUC23 using the restriction enzymes EcoRI and BamHI.
AS substrates are initially added, the activity is higher and then it reaches a plateau with increase. NO. Sucrase activity reached a plateau. Correct Predictions: Optimal temperature: 40 °C (104 °F), Sucrase activity increases with increasing sucrose concentration until a plateau is reached. Incorrect predictions: Sucrase will have highest activity at ph 6 not ph 7 Warm up exercise increases body temperature making it ideal for enzyme activity Increasing myosin activity makes muscles tougher and stiff and limits free movement for performance.
Since the speed of an enzyme reaction relies on whether there is any substrate to react with, float speed should increase as substrate concentration increases. Therefore, if H2O2 concentration increases, float speed will increase. If H2O2 concentration decreases, float speed will decrease. This hypothesis is based off http://www.worthington-biochem.com/introBiochem/substrateConc.html where substrate concentration is seen to increase rate of reaction. It can be noted however that rate of reaction will level off after a certain concentration level, which can be called Vmax, or maximum velocity of reaction.
Another control consisted of 5cm³-distilled water in fifth test tube and 5cm³ casein in a final test tube. All of these were also placed into the water bath. Black card was placed behind the test tubes to help spot the clearing of the solution. The enzyme and substrate were mixed, a stopwatch started immediately, and the time for the suspension to clear noted. This was repeated for the controls, and the whole experiment repeated for different temperatures, ranging from 25°C - 65°C.
Phospholipid bilayers and protein molecules form a semi-permeable boundary around the cell. This is called the cell membrane. Molecules that are small enough, such as water and CO2, can pass in and out of the membranes following the concentration path. In the experiment, dialysis bags will be used as cell membranes, sugar as the solute, and water as the solvent. This experiment is designed to allow us to measure the effect of concentration of sucrose and temperature on osmosis over time.
How does this compare to a real enzyme? The enzyme’s rate did change over time. This compares to a real enzyme because an enzyme’s job is to speed up the reactions and as time allotted. That did happen since the enzyme in our lab was able to make more chainobeads as time progressed. 4.
Investigating the effect of pH on amylase activity This practical allows you to: • discover how pH affects the rate of an enzyme controlled reaction • evaluate the experimental procedure Procedure SAFETY: Follow your teacher’s instructions for handling the solutions. Wear eye protection when handling the iodine solution. Investigation a Place single drops of iodine solution in rows on the tile. b Label a test tube with the pH to be tested. c Use the syringe to place 2 cm3 of amylase into the test tube.
(http://www.ariseandshine.com/enzymes-1.aspx). The purpose of this lab was to design an experiment to demonstrate the action of
Purification and characterization of Cellulobiase By John Bullock Abstract In this experiment we determined the enzyme activity for cellulobiase. Two buffers were prepared in order to quantitate the proteins using ion exchange chromatography. Protein determination was then carried out using linear regression data. Once the protein determination was done we then selected the correct proteins to run our enzyme assay. When measuring the absorbance in this case, we were measuring the product that the enzyme produced.
Enzymes are catalytic proteins that are essential in human metabolism, as they help facilitate many of the biochemical reactions occurring in the body by lowering the activation energy that is necessary for each reaction to occur. Various factors, such as temperature and pH, contribute to the enzyme activity and can be directly regulated to affect the overall rate of reaction. The purpose of this experiment is to determine the optimal physical conditions of the glyocolytic enzyme muscle lactate dehydrogenase (mLDH), which aids in the reduction and conversion of pyruvate to lactate and the oxidation of NADH in skeletal muscle, and to discover the effects of altering temperature and pH on the activity of the enzyme mLDH. Methods In order to complete this experiment and determine the optimum pH and temperature of the enzyme mLDH, 16.2 mM pyruvate in dH2O was placed in 0.2 mM NADH in 54 mM phosphate buffer solutions of varying temperatures and pH levels, in accordance with the following reaction: pyruvate + NADH + H+ lactate + NAD+ In this experiment, 900 µL of NADH solution with a pH 5.5 (at about 25°C) and 25 µL of 1:2000 dilution of muscle homogenate (containing mLDH) were mixed inside a cuvette. After waiting a minute, 50 µL of pyruvate was then added to the cuvette and then mixed.