LABORATORY REPORT Activity: Enzyme Activity Name: Natalie Banc Instructor: Elizabeth Kraske Date: 09.26.2016 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. Independent Variable pH 3. Controlled Variables temperature, amount of substrate (sucrose) present, sucrase + sucrose incubation time Effect of Temperature on Enzyme Activity 1. Dependent Variable amount of product (glucose and fructose) produced 2. Independent Variable temperature 3. Controlled Variables pH, amount of substrate (sucrose) present, sucrase + sucrose incubation time Effect of Substrate Concentration on Enzyme Activity 1. Dependent Variable amount of product (glucose and fructose) produced 2. Independent Variable amount of substrate (sucrose) present 3. Controlled Variables temperature, pH, sucrase + sucrose incubation time 4. Describe what is measured as an indicator of sucrase activity and why this is an indicator of sucrase activity. I believe glucose and fructose was used as an indicator because they are what produces sucrose and sucrose creates more sucrase activity. 5. Explain why denatured sucrase was used as a control. Denatured sucrase was used as a control because it was inactive compared to the active sucrase. Using it slowed
The purpose of this experiment was to analyze the effects of the variables: temperature, pH, and enzyme concentration, on the enzymatic reaction rate of catalase and the level at which its products are released, measuring the rate of absorption using the indicator solution guaiacol and a spectrophotometer to develop a hypothesis of the ideal conditions for these reactions. My hypothesis is that the extremes in concentration, temperature and pH will negatively affect the Au rate. This experiment used 11 solutions contained in cuvettes. Each cuvette, once mixed, is placed in spectrophotometer and then a reading taken every 20 seconds. Cuvettes 1, 8, and 10 are used as blanks to zero out the spectrophotometer.
A control extract is prepared (5ml of DAE) to a test tube, which is then placed in boiling waterbath for 10minutes, after 10minutes remove the control extract and leave it to cool at room temperature. In order to determine the amylase activity, one drop of iodine is dropped into 21 labelled wells on the ceramic test plates. A reaction mixture is prepared, 5ml of buffer and 1ml of 0.5% starch solution to a test tube. Extract one drop from the reaction mixture to the well labelled T.
Throughout this work, a correlation between the enzyme activity and temperature existed. The data for both showed that at the temperature extremities the lower the enzyme activity was; except for the bacterial enzyme at 0°C. At high temperatures, an enzyme denatures or changes shape, making it difficult or impossible for a substrate to bind, and at low temperatures, the frequency and rate of reaction decreases causing for a halt in product formation (Pitzer et.al., 2012). Thus, showing that enzymes need to be in their optimal environments to work properly. Bacillus lincheniformis and Aspergillus oryzae are both organisms that live that in medium temperature environments.
Another factor that can affect the chemical reaction is temperature. It can affect the enzymes by changing their speed and at a certain temperature denaturing them. When enzymes and substrates are moving quickly, they are more likely to collide and have a chemical reaction. The warmer the temperature the faster the chemical reaction is to a certain degree. The chemical reaction was the most effective at 40 because the enzymes and substrates were moving fast, but the temperature was not high enough to denature the enzyme.
A simple change in temperature, a molecule out of place, and a sudden change in the pH level are just some of the things that can harm an enzyme 's reaction rate (the speed at which a chemical reaction proceeds) (5). To test the reaction rate of an enzyme, a lab was done to simulate what would happen to an enzyme under extreme conditions. The enzyme (represented by a hand) had to catalyze as many substrates as possible (represented by toothpicks) within 60 seconds. The experiment dealt with environmental factors such as extreme cold, presence of other molecules, etc. The lab that was simulated directly correlated to many of the topics discussed in class, like explaining the importance of enzymes and measuring the enzymes’ ability to function under different conditions.
The effect of pH on the speed of enzyme interaction with substrate chemicals Hypothesis: About pH: If the pH level is less than 5, then the speed of the enzyme reaction will be slower. About temperature: If the temperature stays the same, then the speed of the enzyme reaction will not be completely affected. Background information: The function of enzymes is to speed up the biochemical reaction by lowering the activation energy, they do this by colliding with the substrate.
In Part A, students would observe the effects of enzyme concentration on the rate of reaction. They were required to make six solutions with different concentration of catalase, the speed of
3. Look at your graph for Part B, how does temperature affect enzyme activity? The colder the temperature the greater the reaction. 4. Look at your graph for Part C, how does pH affect the enzyme activity?
Independent Variable pH 3. Controlled Variables temperature, amount of substrate (sucrose) present, sucrase + sucrose incubation time Effect of Temperature on Enzyme Activity 1. Dependent Variable amount of product (glucose and fructose) produced 2. Independent Variable temperature 3. Controlled Variables pH, amount of
Enzymes speed up chemical reactions enabling more products to be formed within a shorter span of time. Enzymes are fragile and easily disrupted by heat or other mild treatment. Studying the effect of temperature and substrate concentration on enzyme concentration allows better understanding of optimum conditions which enzymes can function. An example of an enzyme catalyzed reaction is enzymatic hydrolysis of an artificial substrate, o-Nitrophenylgalactoside (ONPG) used in place of lactose. Upon hydrolysis by B-galactosidase, a yellow colored compound o-Nitrophenol (ONP) is formed.
Macromolecules are usually used to refer to large biological polymer which are made up of small monomers linked together. All living things contain organic macromolecules, which is divided into four main groups: Lipids, proteins, carbohydrates and nucleic acids. (D 'Onofrio, 2009-2015) Characteristic for these organic molecules is that they are made up of only a small number of elements: carbon, hydrogen, oxygen, and to smaller amounts nitrogen, phosphorus and sulfur. Carbohydrates are better known as sugars and starches.
If the conditions are not desirable for the enzyme, it will perform at a slower rate, or denature. For both enzyme and substrate concentration, an increase in concentration will result in increased activity until the optimal point. Enzymes tend to perform best at higher temperatures, until reaching their optimal point. After this point their behavior rapidly worsens until they denature. Enzymes also have an optimal pH range.
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).
Amylase hydrolyses (breaks down) starch and glycogen into more simple and readily digestible forms of sugar (glucose). Commercially available Amylase solutions can be easily used to breakdown complex carbohydrates (e.g. starch) into simpler forms of sugars (e.g. disaccharides and monosaccharaides). Copper Sulphate can block the activity of Amylase, which is a known non-competitive irreversible enzyme inhibitor. The light absorbent method can be used to study this phenomenon of breakdown and blockade of breakdown of starch in the laboratory. After studying these properties of Amylase and Copper sulphate I designed my experiment to study the inhibitory effect of Copper Sulphate on the enzymatic activity of 1% and 2% Amylase solution.
ABSTRACT: The purpose of the experiments for week 5 and week 6 support each other in the further understanding of enzyme reactions. During week 5, the effects of a substrate and enzyme concentration on enzyme reaction rate was observed. Week 6, the effects of temperature and inhibitor on a reaction rate were monitored. For testing the effects of concentrations, we needed to use the table that was used in week 3, Cells.