Enzymes Lab Report

15) Obtain the boiled chloroplast suspension, mix, and transfer 3 drops to cuvette 4. Immediately cover and mix cuvette 4. Insert it into the spectrophotometer's sample holder, read the percentage transmittance, and record it in Table 4.4. Replace cuvette 4 into the incubation test tube rack. Take and record additional readings at 5, 10, and 15 minutes. Mix the cuvette's contents just prior to each readings. Remember to use cuvtte 1 occasionally to check and adjust the spectrophotometer to 100% transmittance.

According to Figure 2, Tube 3 and Tube 4 were very similar. They display a significant increase from the initial and final recordings. These tubes show that lower salt concentration is optimal for enzyme activity. Due to the lower concentration of NaCl in the solution, the enzyme is able to react more with the substrate. The more the ONPG reacts with the Lactaid, the more product is created, which in turn makes the solution a more concentrated yellow rather than a pale yellow.

In the experiment, it was proven that Tube 13 had the highest enzyme activity condition. The question being asked in this experiment is how does substrate concentration and pH affect enzyme function. It was hypothesized that there will be an optimal condition outside which function is lower. The group predicted that optimal conditions are pH is 7 and substrate concentration is 6 percent. In conclusion, this whole experiment justifies that enzymes are affected by the changes in pH. Where enzymes are most active is where you will find the most favorable pH value. According to the data, the substrate concentration with the highest enzyme activity is the 0 percent H2O2, which has a value of 0.0093. When compared to literature the 6 percent H2O2

The tubes sat at room temperature for 10 minutes, and they were inverted and shaken every 2 minutes in order to mix the enzymes and chelating agents. They sat at room temperature for 10 minutes while being mixed every 2

For the first experiment the objective was to measure the disappearance of substrate and the amount of product being produced. The product in the experiment is benzoquinone which is a reddish brown color that is easy to see how much is being made. In order for the reaction to happen, it must have a substrate and enzyme. The substrate is catechol and the enzyme is catecholase in the experiment. The first step was to get three clean test tubes. We then had to zero an instrument called a spectrometer used to measure light absorbency. The instrument was zeroed by using a blank. For test tube B we measured 1 mL of potato extract and 5mL of distilled water to put inside. After we tightly covered test tube B with Parafilm in order to mix the contents together. On the Spec 20 we set the wavelength to 540 nm and also using the knob to the left we set the absorbency to stay at infinity. After we carefully wiped the tune and placed it in the sample holder of the Spec 20. Then on the knob to the right we set the absorbance to zero. Now that the spec 20 is zeroed, the tubes are ready to be measured because of the catecholase reaction taken place. We then used a pipet to measure 3 mL of water in tube 1,

Competitive inhibitors bind to the active site preventing the binding of substrates, noncompetitive inhibitors join another part of the enzyme and distort the active site so that the substrate molecule is no longer complementary. Cofactors are required as they loosely bind to a protein and are required for the proteins reaction to occur (Proprofs,

The same experiment was repeated for part C of the lab, however, three different data sets were recorded. The first data set contained the addition of DMSO, a second data set had the addition of the inhibitor quercetin, while the third data set contained fruit extract. The concentration of tyrosinase, quercetin, DMSO, and fruit extract were constant, while concentration of L-DOPA and phosphate buffer differed from each cuvette as recorded on pages 22 and 23 of the Lab Handout. One modification to part C compared to part A and B is that the volumes of each reagent was cut down by a third due to supplies running low, however, this did not affect the experiment in any ways. As seen on pages 22- 25 on the lab manual, after linear plots and Michaelis-Menten graphs

Benedict’s reagent is a bright blue solution that changes to green to orange to reddish-brown with increasing amounts of maltose. It is important to understand that enzyme assays only indicate the presence or absence of substances. It is up to you to analyze the results of the experiments to decide if enzymatic hydrolysis has occurred. Choose Chemical and Physical Processes of Digestion from the main menu. The opening screen will appear in a few seconds (Figure 8.1). The solutions shelf in the upper right part of the screen contains the substances to be used in the experiment. The incubation unit beneath the solutions shelf contains a rack of test tube holders and the apparatus needed to run the experiments. Test tubes from the test tube washer on the left part of the screen are loaded into the rack in the incubation unit by clicking and holding the mouse button on the first tube, and then releasing (dragging-and-dropping) it into any position in the rack. The substances in the dropper bottles on the solutions shelf are dispensed by dragging-and-dropping the

Background Information: Another word for enzymes is catalysts and most of them are proteins. They bind to the substrates(reactants) temporarily of the very reaction that they catalyze. This speeds up the chemical reaction because by doing this, the enzymes lower the amount of the activation energy required to start the reaction. In the active site the the substrates are held by forces that are noncovalent who are an assortment of hydrophobic reactions, hydrogen bonds, and ionic interactions. Most of these interactions are weak and they are for sure weak if the atoms involved in the reaction are more than one angstrom from each other.

The purpose of the enzyme lab conducted was to observe the chemical composition of cells. In order to do so we tested for the presence of organic molecules. Molecules are what forms when atoms bond together. Organic molecules of cells include proteins, carbohydrates, and lipids, which are composed of smaller molecules known as monomers and polymers. Polymers are joined monomers. A chemical reaction links monomers together occurs and releases a water molecule, this is called dehydration synthesis. Hydrolysis separates polymers into monomers by using water to break bonds. Organic catalysts called enzymes are proteins that increase the speed of a chemical reaction. In the lab we used Biuret reagent to test for proteins, iodine solution to

Enzyme activity also depends on the concentrations of enzyme and substrate. As substrate concentration increases in a solution, enzyme activity will also increase due to the creation of enzyme-substrate complexes. However, once all the enzymes have been saturated with substrate, their activity cannot increase anymore. At this point, commonly known as the Vmax point, enzyme activity tends to remain at a steady value despite any more increase in substrate concentration.

To begin our experiment, we needed five cuvettes, two flasks, two beakers, glucose, sucrose, lactose, phosphate buffer and ortho-nitro-phenyl-galactoside (ONPG), a lactase pill (our enzyme) and a spectrophotometer.

Beta lactamases are enzymes produced by bacteria that give bacteria resistance to beta lactams, the most commonly prescribed class of antibiotics, such as cephalosporins and carbapenems. They act by hydrolyzing the four-member lactam ring of beta lactams, rendering them inactive.

The first lab preformed was a harvest assay. First a plaque was picked from the last titer assay. This plaque was then added to an epindorf tube with 100 microliters of phage buffer in it. A serial dilution was then done to 10 ^-3, 10 microliters of each dilution was added to. 0.3 milliliters of M. Smegmatis. The solution was then incubated at room temperature for fifteen minutes. Then 4.5 milliliters of top agar was added to each tube and plate onto petri dishes. Next the most diluted plate was flooded with 4 milliliters of phage buffer, incubated at room temperature for two hours. The plate was then inverted in order to collect one milliliter of liquid collected on the plate’s top, which was then placed in an epindorf tube. From that epindorf tube 10 microliters of it was then added to 90 microliters of phage buffer and vortexed. A serial dilution was then completed to 10 ^ -4. Each dilution was then added its own tube with 0.3 milliliters or M. Smegmatis. To those tubes 4.5 milliliters of top agar was then added to each and each were plate on to their own petri dish. The 10^-3 and 10^ -4 plates from the previous harvest assay were used in the next harvest assay. Both plates, the 10^-3 and 10^-4, were flooded with 4 milliliters of phage buffer each. These plates were then incubated for two hours. After the two hour incubation period the

First put on your goggles, gloves, and lab coat. Before measuring out all the enzyme and substrate concentrations we have to plug and turn on the spectrophotometer because it takes about 20 to 30 minutes to warm up. While the spectrophotometer is warming up we will start to prep the test tubes for the enzyme concentration on the reaction rate. We will have a low, medium and high concentration and for each level of concentration we will have a control, substrate, and enzyme. Starting with the low enzyme concentration, the configuration of the control should contain: 0.1 mL of guaiacol, 0.5 mL of turnip extract, and 9.4 mL of distilled water which should equal to