...BIOL 3380 Name:_____________________________________ Circle Session: T-PM W-AM W-PM R-AM R-PM F-AM F-PM Experiment 9 – Pre-lab Homework Enzyme Kinetics of LDH This pre-lab homework assignment is due at the beginning of your lab session. You are provided with the following portion of a protocol: • Determine concentration of enzyme stock solution, if unknown, by taking an A280 nm reading of a 1:100 dilution (in water). Use a total volume of 1 ml in the cuvette. • Dilute some of the enzyme stock with buffer A to make a 4 mg/ml solution. • Serially dilute the 4 mg/ml solution with buffer A to make working solutions of 400 µg/ml and 40 µg/ml. • Prepare 30 µl of each working solution for every sample The PI of the lab gives you a tube of enzyme and tells you the following before disappearing into the office to write more grant proposals: ➢ There is 50 µl of enzyme stock solution. The enzyme is expensive to purify, so follow the protocol exactly, using as little of the stock solution as possible. ➢ The concentration of the stock solution is currently not known, but a 1 mg/ml concentration of the pure enzyme has an A280 nm of 2.0. ➢ You’ll be performing the assay on 12 samples. ➢ Make enough of each working solution so that you have at least 400 ul to work with when you do the assay (to cover any waste and/or inefficiencies in pippetting). Using the spectrophotometer to read the absorbance at 280 nm, you get...
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...Enzymes: Virtual Lab Introduction: This is a virtual lab simulation of enzymes and substrates. It teaches about enzymatic activity and how it is affected by substrate concentration and pH. Students are to visualize the virtual lab as an actual lab and conduct the experiment as such. Purpose This investigation will determine the effects of substrate concentration and pH on the initial rate of an enzyme-catalyzed reaction. Materials Computer Pencil/pen Enzymes at various pH Substrates at various concentrations Procedures 1. Go to this link ( http://www.mhhe.com/biosci/genbio/virtual_labs/BL_11/BL_11.html 2. Click the TV/VCR. Then click the play button. Watch an animation about enzymes. 3. Click Information and read more about enzymes and substrates. 4. Complete the table found at the bottom of the virtual lab by: a. Adjusting the pH level of the test tubes (already filled with an enzyme solution) by clicking the up and down arrows. Pay attention to the proper pH in each designated test tube according to the data table! b. Adding substrate to each of the test tubes that already contain an enzyme solution by clicking and dragging a piece of weighing paper with the powdered substrate. Pay attention to the proper substrate concentration in each designated test tube according to the data table! c. Record the number of molecules of product formed per minute into your data table. d. Click the computer monitor to see...
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...Enzyme Catalysis: Lab Report Introduction: In the lab there were 4 parts (A, B, C, D). Each part having a different reaction among it, an example of this reaction could be lab A when the catalysis was tested for activity. The reaction was to support the affect catalysis had on the hydrogen peroxide. This was a fizzing and bubbling state supporting a reaction was occurring. So one can infer that if enzyme catalysis brakes down hydrogen peroxide, then an abandons of catalysis would break down more hydrogen peroxide upon a quicker rate. Materials and methods: Part A: hydrogen peroxide, catalysis. After taking 10ml of H2O2 and mixing it with 1ml of catalysis for 30 seconds. After the reaction the data was recorded. Part B: H2H2, H4SO4, KMnO4 H2O. In the reaction an observation of 10ml of H2O2 in a 60ml container, next 1ml of distilled water is transferred, then 10ml H2SO4 is moved to a cup labeled baseline, finally 5ml of the solution is moved from baseline to titration and is titrated. Lastly the data is recorded. Part C: H2O2 (overnight), H2SO4, KMnO4. Basically the same reaction that occurred in part B was duplicated, but with an overnight solution of H2O2. It was in a baseline and the transferred to titration and then data was recorded. Part D: H2O2, H2SO4, KMnO4. With 60ml cups labeled 10sec, 30sec, 60sec,120sec, and 180sec catalysis was put to a decomposes test. 1ml of catalysis was transferred to a cup and then was swirled. Finally H2SO4 was added to stop the enzyme...
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...What Influences Enzyme Activity Biology Lab 2010-09-25 Summary In this lab we learned about what influences enzyme activity. We learned many terms and concepts in this lab. Enzymes decreases the amount of energy needed in a reaction. Catalyst speeds up reaction. A substrate is what the material with which catalyst reacts. A product is the modification of the substrate. This was a very informative and good lab. Materials 1. 1 Reaction spot plate 2. 3 Small Cups 3. 3 Plastic pipettes 4. 1 Bottle starch indicator solution. 5. Prepared starch solution 6. Prepared diastase solution 7. Distilled water 8. Clock with second hand 9. Bottle of dilute hydrochloric acid (HCL) solution- 0.1 % 10. Bottle of dilute sodium hydroxide (NaOH)- 0.1% 11. Test tubes 12. 2 Glucose test strips 13. Glucose Test Strip Color Chart 14. Clock or Stopwatch Procedure: Activity 1- Effect of Enzyme Concentration on Activity 1. Obtain approximately 10 mL each of the prepared starch solution, the diastase solution, and distilled water and place each of them in one of the sample cups. Label each of your solutions properly. 2. Using two different plastic pipets, place one drop of enzyme in each of 12 successive wells on the spot plate, followed by four drops of distilled water. Quickly put one drop of starch solution in each of the wells using a third pipet. Be...
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...BIOL160 (title of lab report) Lab 3 26 February 2013 Lab partners: Hypothesis If the time of starch disappearance is directly related to the temperature, then the higher the temperature the more active the amylase is until it denatures and becomes non-functional. Results Tube | Temperature (c) | Time of starch disappearance (mins) | 1 & 1A | 80 | | 2 & 2A | 70 | t= 3 | 3 & 3A | room temperature | t= 2 | 4 & 4A | ice slurry | | 5 & 5A (human saliva) | room temperature | t= 3 | 6 & 6A (human saliva) | 37 | t= 1 | Discussion Based on the analysis shown, the hypothesis appears to be proven correct. The higher the temperature the more active the amylase is until it denatures and becomes non-functional. This is due to the exceeding time on the reaction of enzyme. The enzyme either has been denatured or its reaction time is extremely slow. It is encouraged that this experiment should be repeated a number of times to get a confirmation on the results as a proof that the data and the hypothesis set is not a coincidence. In this experiment, we did not encounter any denaturation of the enzymes. However, it would also be interesting to find the point of denaturation; this would be achieved by performing the experiment again with environmental conditions between 22°C and 37°C, as it appeared as though the denaturation...
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...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 test for starch, paper to test for lipids. In the first lab, we tested for the presence of proteins in samples by using blue solution called Biuret reagent, which changes to purple when a protein is present and pinkish-purple for peptides. First test tubes were marked at 1cm and then filled to the mark with water, albumin, pepsin, and starch. Next, five drops of Biuret reagent was added to the sample, covered with Parafilm, and swirled to mix. The water remained clear, indicating the sample lacked the presence of proteins, and thus was our negative control. The albumin sample observed changed to an orange-purple color, indicating the presence of protein. The peptin sample changed to a pink-purple hue, testing positive for presence of peptides...
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...Introduction Enzymes are protein molecules that catalyze chemical reactions in all living organisms. Enzymes allow living organisms to carry out complex chemical activities at low temperatures, but can’t cause a reaction that hasn’t occurred in their absence. Also, enzymes are thought to speed up reactions by bringing reacting molecules together to increase the chances that a reaction will occur (Worthington Biomedical Corporation, 2015). Each enzyme has a specific active site where the substrates attach. Many factors can affect enzyme activity such as temperature, pH, and the presence of inhibitors (John W. Kimball, 2014). The purpose of this lab was to examine factors affecting the enzyme function of peroxidase. In the 19th century French chemist Louis Jacques discovered catalysts. Catalysts are substances that enable a chemical reaction without participating in it, which led to specifically peroxidases. The structure of peroxidase is a very large enzymatic protein, and has complex molecules with complicated shapes involving multiple folding’s. The activity of peroxidase is dependent on pH. It exhibits maximum activity at a pH between 6.5 and 7.0. The activity of the enzyme is reduced when pH levels are increased. Peroxidase promotes the oxidation of various compounds naturally of peroxides, where hydrogen peroxide is reduced to form water (Wikimedia Foundation, 2015). Also peroxidases break compounds down into harmless substances by adding donor molecules. During this lab, the donor...
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...DNA Fingerprinting Using Agarose Gel S. Aaron Sowards Bio 122 Lab 04 Brianna Adanitsch Jakob Lester Minhenga Ngijoi 2/21/18 Dr. Chad R. Sethman Abstract DNA fingerprinting is the process of analyzing an individual’s DNA base-pair patterns. The DNA fingerprinting lab involved identifying the suspect using Agarose Gel and Polymerase Chain Reaction. It was found that suspect two s DNA matched the crime scene DNA. This is known because suspect twos DNA traveled the same distance as the crime scene DNA. DNA Fingerprinting Using Agarose Gel Introduction In 1984 Dr. Alex Jeffreys came up with deoxyribonucleic acid (DNA) fingerprinting, which is also known as DNA profiling or DNA typing. DNA fingerprinting is the analyzing...
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... Enzyme lab report Determining the properties of an Enzyme Introduction: Enzymes are proteins that acts as catalysts for reactions. This means that enzymes lower the activation energy essential for a reaction to take place, allowing a specific reaction to occur much quicker and easier. Certain enzymes only lower the activation energy for certain reactions, and enzymes are shape precise. The distinctive folds of the amino acid chains that make up an enzyme result in the formation of a precisely shape active site. When the reactants of a reaction, substrates, fit seamlessly into the active site of an enzyme, the enzyme is then able to catalyze the reaction. The activity of enzymes is affected by the concentration of enzymes current and the concentration of substrate current. As the amount of enzyme present increases, the rate of reaction increases. Most enzymes need specific environmental conditions to be met in order for them to function properly and efficiently. These conditions include the pH level, temperature, and the inhibitor. If the ideal conditions for an enzyme are altered, the enzyme may denature, or change its shape, resulting in deactivation. As a result, the enzyme activity would be that it would no longer be able to catalyze the reaction, and the reaction rate would significantly decrease. Methods and Materials: * Temperature...
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...Lab Report #2 Name: Lab: #9 Enzymes – Experiment #4 Due date: Purpose The purpose of the experiment is to compare and examine the effect of substrate concentration on catalase activity. Introduction All chemical reactions require a catalyst. A type of catalyst that exists is an enzyme, which acts to bring out a specific biochemical reaction. At all times, all work inside a cell is being performed by enzymes (Brian, 2000). The purpose of an enzyme is to help the cell carry out reactions very quickly. An interaction must be made for a reaction to become catalyzed. The active site is where this interaction between the enzyme and the reactant and/or reactants takes place. In order for the enzyme to work efficiently and properly, the reactant (or substrate) must position itself perfectly within the active site. Most enzymes usually only can catalyze a single chemical reaction, which is called specificity (Introduction To Enzymes, n.d.). Enzymes can also operate to an optimal extent where chemical reactions can occur rapidly and with the upmost efficiency, under certain conditions known as the enzyme’s optimum activity (Boli, 2012). The many different conditions include environmental, such as pH and temperature, or concentrations of the substrates and enzymes. In this experiment, we examined a substance called catalase. Catalase is the isolated cells from potatoes and beef liver. As the substrate for the experiment, hydrogen peroxide was used at various different amounts...
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... |Liow Yi Sheng | | |Foo Yong Hao | |Practical Group |P13 | |Date of lab class |13/7/2015 | |Program |Foundation in Science | |Unit code |FHSB1214 | |Unit description |Biology I | |Year and trimester of study |2015, Trimester 1 | |Title of lab report |Investigation of the effects of different catalytic conditions on hydrogen peroxide | | |decomposition | |Lecturer’s name |Ms.Ting Jen Ching...
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...What is the function of enzymes in a living system? Enzymes speed up metabolic reactions necessary for life. Without them certain vital processes would not take place and the body would be unable to function. Difference enzymes work better under different conditions. Where in a human body might it be beneficial to have enzymes work in very acidic environments? In areas, like the stomach, that have a pH of two would benefit by having enzymes that function well in acidic environments. An example of such an enzyme is pepsin. There is a large amount of catalase found in a human liver. Does the liver break down more hydrogen peroxide in the summer or winter? Explain your answer. More hydrogen peroxide will be broken down in the summer compared to the winter because higher body temperatures equals more enzyme activity. Many enzymes end with “ase”. Come up with your own enzyme, then name and explain what this enzyme does. Draw the enzyme and the substrate in the space provided below along with the enzyme-substrate complex. My enzyme would be olestrase. It would break down the lipid olestra and make it usable for the human body. Recent advances have allowed humans to mass-produce certain enzymes. Research one such enzyme and explain how this enzyme has been used to benefit society. Coenzyme Q10 (ubiquinone) is a naturally occurring substance which has properties potentially beneficial for preventing cellular damage during myocardial ischemia and reperfusion. It plays...
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...Marlena Ferone LeafLab Biology 101 - Professor Landry University of Phoenix Due Date: August 30, 2011 EXPLORATION What is the relationship between the increase in light intensity and the photosynthetic rate in leaves from a corn plant? How does this relationship compare with what you observed for tomato plants? As light intensity increases, the photosynthetic rate of a plant increases as well. However biochemical reaction, in fact, DO have temperature limits because if the temperatures reach a certain degree and overheat, the enzymes will get damaged and/or lost and inevitably die. Therefore, temperatures must be a tolerable limits in order to increase a photosynthetic rate. Photosynthesis at low temperatures actually take CO2 more efficiently. The rate of photosynthesis is decreased by higher oxygen concentrations. Lower temperatures inhibit Rubisco and forces this enzyme to to work competitively. Rubisco works competitively because is actually binds itself to oxygen rather than binding to carbon dioxide because of the mere attraction. Therefore, in doing so, this activates photorespiration rather than photosynthesis. Photosynthesis in corn is different from photosynthesis in a tomato. Corn is part of CO4. CO4 plants are actually able to capture CO2 into precursor acids. In doing so, CO2 is kept at a constant rate. Increases and decreases in temperature do not affect the rate of CO2. Corn is able to continue efficient energy production which surprisingly allows it...
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...LAB REPORT: ENZYMES Part I: Graphs and Data TIME COURSE: ABSORBANCE VS. TIME Provided data: Time(minutes) | Experimental ABS @ 405nm | Control ABS @ 405 nm | Exp. ABS minus Control ABS | Micromoles p-Nitrophenol | 0 | 0.057 | 0.051 | 0.006 | 0.0004 | 10 | 0.207 | 0.053 | 0.154 | 0.0064 | 20 | 0.351 | 0.054 | 0.297 | 0.0120 | 30 | 0.501 | 0.055 | 0.446 | 0.0181 | 60 | 0.955 | 0.064 | 0.891 | 0.0362 | Personal data: Time(minutes) | Experimental ABS @ 405nm | Control ABS @ 405 nm | Exp. ABS minus Control ABS | Micromoles p-Nitrophenol | 0 | 0.092 | 0.064 | 0.028 | 0.0010 | 10 | 0.262 | 0.048 | 0.214 | 0.0085 | 20 | 0.429 | 0.054 | 0.375 | 0.0140 | 30 | 0.599 | 0.051 | 0.548 | 0.0208 | 60 | 0.976 | 0.050 | 0.926 | 0.0350 | STANDARD CURVE: Provided data: Micromoles p-Nitrophenol | Absorbance @ 405 nm | 0.0000 | 0.000 | 0.0025 | 0.058 | 0.0050 | 0.118 | 0.0100 | 0.245 | 0.0200 | 0.496 | 0.0400 | 1.000 | Personal data: Micromoles p-Nitrophenol | Absorbance @ 405 nm | 0.0000 | 0.000 | 0.0025 | 0.071 | 0.0050 | 0.167 | 0.0100 | 0.228 | 0.0200 | 0.519 | 0.0400 | 1.050 | TIME COURSE: PRODUCT VS. TIME Provided data: Time | Micromoles product | 0 | 0.0004 | 10 | 0.0064 | 20 | 0.0120 | 30 | 0.0181 | 60 | 0.0362 | Personal data: Time | Micromoles product | 0 | 0.0010 | 10 | 0.0085 | 20 | 0.0140 | 30 | 0.0208 | 60 | 0.0350 | TEMPERATURE: PRODUCT VS. TEMPERATURE Provided data: ...
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...Torres Group #11 USC Chemistry 322b Formal Lab Report 6th November 2015 I. II. Enzymatic Resolution of 1-Phenylethanol and Diastereomer Analysis Objective/Abstract Enzymatic transesterification reaction was performed to study the resolution of diastereomers using 1H-NMR analysis. The stereo-selectivity of acylase I, an enzyme, for a 50:50 racemic mixture of 1-phenylethanol was determined. In the first of a two-step reaction, 1-phenylethanol was reacted with vinyl acetate with the help of acylase I to form an ester, unreacted 1-phenylethanol, and vinyl alcohol. The unreacted 1phenylethanol was separated from the ester by column chromatography and confirmed by thin-layer chromatography (TLC). In the second reaction, the unreacted 1-phenylethanol was reacted with (R)-(-)-acetoxyphenylacetic acid to form a diastereomer ester. In the latter reaction, four different 1-phenylethanol samples were used in order to compare 1HNMR data of the resulting diastereomer esters and determine which enantiomer of the 50:50 racemic mixture was preferred by acylase I. Those four samples were: (1) racemic 1-phenylethanol, (2) unreacted 1-phenylethanol, (3) (R)-1-phenylethanol, and (4) (S)-1phenylethanol. After 1H-NMR analysis, it was found that the (S)-1-phenylethanol was preferred by acylase I. III. Introduction and Background Information Scheme 1: Reaction of 1-phenylethanol with vinyl acetate in the presence of the enzyme acylase I to produce 1-phenylethyl acetate (ester)...
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