Investigating The Amount Of Enzyme That Would Produce A Reaction Rate

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

Used to see if the temperature of the water is at 37oc – 40oc and if

Lab six requires students to observe the effects of pH and enzyme concentration on catecholase activity. Enzymes are organic catalysts that can affect the rate of a chemical reaction depending on the pH level and the concentration of the enzyme. As pH comes closer to a neutral pH the enzyme is at its greatest effectiveness. Also at the absorbance of a slope of 0.0122 the enzyme is affected greatly. The pH effect on enzymes can be tested by trying each pH level with a pH buffer of the same pH as labeled as the test tube and 1mL of potato juice, water, and catechol. This is all mixed together and put in the spectrophotometer to test how much is being absorbed at 420nm. As the effect on enzyme concentration can be tested almost the same way. This part of the exercise uses different amounts of pH 7-phosphate buffer and potato juice, and 1mL of catechol mixed together in a test tube. Each substance is put in the spectrophotometer at a wavelength set tot 420nm. The results are put down for every minute up to six minutes to see how enzyme concentration affects reaction rate. The results show that the pH 8 (0.494) affects the enzyme more than a pH of 4 (0.249), 6 (0.371), 7 (0.456), and 10 (0.126). Also the absorbance is greatest at a slope of 0.0122 with test tube C that has more effect on the reaction rate, than test tube A, B, and D.

PURPOSE: Measure the effects of changes in catalase concentration, substrate concentration, and salinity on the reaction rates of an enzyme.

The purpose of this lab is to test for enzyme activity by examining factors that may influence enzymes.

The experiments were performed in the science lab 1.226 at the University of Texas Rio Grande Valley, Edinburg on October 2, 2017. The experiments were performed in a two-day process due to lack of time. Instructions were given by our TA on where to find the substances (guaiacol under the fume hood, turnip extract, peroxide, and distilled water were placed on our lab tables in dropper bottles, along with the spectrophotometer) and were told to get started. In activity 1 we will be testing 3 concentrations of an enzyme (0.5 ml, 1.0 ml, and 2.0 ml of turnip extract). To quantify the rate of reaction in turnips, guaiacol will be used as the color reagent. Guaiacol is oxidized when it encounters peroxide, allowing light at 470 nm to be absorbed and allowing us to measure the absorbance. In the first activity from experiment day 1, three test tubes were obtained and two clean cuvettes from our lab TA, and placed in a test tube rack on our lab tables. We used one of the test tubes to make the control, another to make the substrate and the last one to make the enzyme. We did this process 3 times to test the effects of the low enzyme concentration, medium enzyme concentration, and high enzyme concentration on the enzyme reaction rate. For the low enzyme concentration, on the control test tube we added 1.0 ml of guaiacol, 0.5 ml turnip extract, 0 ml of peroxide and 8.5 ml of distilled water, getting a total volume of 10 ml in the test tube. For the low enzyme concentration, on the

The two experiments investigated measure the activity of the enzyme, catechol oxidase. Specifically, they investigate the effect of decreasing amounts of enzyme on rate of reaction and effect of decreasing amounts of substrate on rate of reaction. Enzymes are proteins with specific structures determined by the sequence of amino acid used to accelerate and regulate biochemical processes; enzyme activity can be measured using the rate at which the reaction catalyzes and can be expressed in concentration of substrate or product. Catechol oxidase, the enzyme used in the experiments, can be found in potatoes and catalyzes the oxidation of catechol to ortho-quinone. This substance turns fruits brown when cut open. The ortho-quinone produced in the experiments was used to determine the reaction rate. The experiments involved enzyme dilutions mixed with water and catechol in experiment II and .026M catechol with water and potassium phosphate in experiment III. Both experiments were measured using a spectrophotometer. My hypothesis for experiment II stated that as the enzyme concentration increased the rate of reaction would also increase at a constant rate, and my hypothesis for experiment III stated that the substrate concentration would increase as the rate of reaction increased at a constant rate. The results concluded that in experiment II the enzyme concentration increased at a constant rate, while the reaction rate increased. The experiment III concluded that the

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

Enzymes are the most important types of proteins, they act as catalysis (speed up chemical reactions). If enzymes didn’t exist, biochemical reactions would act to slowly and they couldn’t keep up with the metabolic functions. Enzymes have a three-dimensional structure that is really complex. This structure consists of one or more polypeptide chains, they form an active site, which is an area in which the substrate eventually will fit. The four factors that affect the activity and reaction rate of an enzyme are temperature, pH, enzyme concentration and

As the chemical reaction progresses, the presence and production of oxygen will determine the rate of the enzyme’s activity through the process. The rate of reaction of the enzyme set with every different temperature will be acquired by measuring the amount of oxygen produced and present within the test tubes in terms of the height its foam in millimeters within each trial. If the amount of foam or the oxygen gas formed appears to be large, it will indicate that the rate of the enzyme’s activity is relatively rapid. Likewise, if the the amount of oxygen appears to be relatively fall under a lower range of foam height, it will indicate that the rate of the enzyme’s activity is slower. Nevertheless, because this enzyme is a form of protein, it is appropriate to indicate that it is very fragile.

This experiment was carried out with many precautions to minimise error margins. These measures included putting the measuring cylinders in the water bath/ ice box/ and bench top at the same time, using the same capsicum extract, having the same person record the results and add the capsicum extract. However there were still some results that exhibited error such as collecting the measuring cylinders at different times which prolonged the exposure, as well as have a delay between starting the timer and adding the capsicum extract. Other issues consist of having an uneven layer of foam (O2) which was a random error that made it difficult to record mL, and forgetting to record the temperature of the H2O2.

In the comparison of the two reactions occurring with the 80% in part A and B it makes sense that the rate of reaction in part B is slower because of the non-competitive inhibitor that was acting upon the catalase. But, the rate of reaction seems to be too low for the result too be realistic indicating a human error and causing it to be slightly unreliable. Failing to change the hydrogen peroxide from part A to a new solution in part B may be an example of human error. An experimental error that may have affected the results obtained during the experiment was the labs failure to mention to mix the two substances, hydrogen peroxide and copper II sulphate in part B. Failure to mention this simple instruction may have led to the outliers in time retrieved in part B for the experiment. In the first trial when the filter timed out at 240 seconds at the bottom of the beaker, it may have been caused by the un-dissolved copper II sulphate still at the bottom of the beaker. Another experimental error was that during the experiment it was stated that if one was able to hurry and do the first two trials quickly then there may be time for a third

The reaction rate of enzymes decreases with salt due to ions present in the water. These ions are so strong that they block the interactions between amino acids and denature the protein or the ions in saline are present in such a minute quantity that enzyme attract each other and becomes inactive which inhibit the enzyme activity by changing the shape of an

Tyrosinase and DOPA are key elements in making melanin. In order to model this reaction, we obtained the enzyme from a 15 g peeled potato and placed the chilled enzyme into five pairs of tests tubes with different pHs to measure the amount of melanin produced. The blank test tubes consisted of 4.9 ml of pH 6.0 phosphate buffer in the blanks, 0.1 ml tyrosinase extract, and a pH of 3,5,7,9 or 11. Each reacting test tube consisted of 3.9 ml of pH 6.0 phosphate buffer, 0.1 ml tyrosinase extract, pH of 3,5,7,9 or 11 and 1.0 ml of DOPA. Each test tube was measured with a spectrophotometer at a wavelength at 450 nm at time 0 and every two minutes for fourteen minutes. A blank was put in before measuring each test tube with the DOPA. After

The purpose of this lab is to give researchers and students the opportunity to design an experiment that determines the effects of various factors on the effectiveness of an enzyme. For this lab, the factor that was chosen to be investigated was pH on an enzyme called peroxidase. The enzyme used in this experiment was (Approximately: 2g) of blended calf liver and could quickly show the conversion of peroxide to water and oxygen. Different bases and acids are important so that people can see how much water and oxygen can be produced. The problem of this lab would be knowledge on things like how to design and understand the steps of a created experiment and get the results hoped for. For this lab the researcher would hope to expect 2H2O2---PeroxidaseO2 + 2H2O.