Bio Chem lab Report 04 Enzyme Biochemistry Group Member: Chan Man Jeun Duncan (16002621) Law Sze Man (16000478) Introduction Enzyme is a protein base structure substance in our body. It works at a biocatalyst that will catalyzing the chemical reaction, which helps to speed up the chemical reaction. Enzyme could only function in specific shape, and the shape of enzyme is depending on the environment, therefore it is hard for an enzyme to function well in an extreme environment. The aim of this experiment is to see can the enzyme functions normally in different environment(pH, temperature and salt concentration) via using starch solution, amylase from saliva, 0.5M HCl solution, 0.5M NaOH solution and NaCl solution, and using iodine solution
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.
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.
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.
Enzymes are biological catalysts, which are essential for carrying metabolic reactions in the human body including the breakdown of food for digestion, absorption and energy production. All biological reactions within human cells depend on enzymes (Wolfenden 1). It is essential for humans to have well-functioning enzymes to break down large molecules into smaller units. As a matter of fact, in the absence of normal functioning enzymes, the human body would cease to exist because chemical reactions that are required to maintain the body function would not occur fast enough. I have a lot of interest in health and human nutrition. Therefore I wanted to examine the breakdown properties of a digestive enzyme while under the influence of a strong inhibitor. For my experiment I chose amylase as an enzyme and starch as a substrate (which is broken down into glucose by Amylase). I selected Copper Sulphate as enzyme inhibitor against the concentration of 2% of Amylase solution. Light absorbance was the method used to
The percentage of glucose was recorded for each sample. Next, the test tubes were carefully cleaned with soap and water. Then five millilitres of sample “A” was placed in the test tube labeled “A”. This was then repeated with the next three samples. 20 drops of Biuret reagent were then added to each test tube.
Starch solution is then placed into the test tube at a quantity of 5 mL. 5 drops of Lugol’s Iodine solution is added to the test tube. If the color changes, then it is known that starches are present in the solution. Proteins are next tested. In order to do this, 5 mL of gelatin solution is added to the test tube. 10 drops of Biuret’s reagent are added to test for protein.
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.
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. The 3 concentrations of enzymes were 0.5 ml, 1.0 ml, and 2.0 ml of turnip extract, while the substrate consisted of 0.1ml, 0.2 ml, and 0.4 ml of hydrogen peroxide. In a separate tube, the control was made up of turnip extract and guaiacol, known as the color reagent. This was recorded the absorbance every 20 seconds for 3 minutes.
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. They all lack the enzyme to help determine the absorption of just the enzyme.
LABORATORY REPORT Activity: Enzyme Activity Name: Natalie Banc Instructor: Elizabeth Kraske Date: 09.22.2016 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
The purpose of this lab is to determine the relationship that exists between the number of amylase gene copies and ancestral diet. As the human civilization moved forward toward agriculture the diets of humans also changed. Depending on where the humans originated would give insight to how much of their diet was starch based. My family’s geographic origins are from China. Thus knowing that the country has a high starch based diet, we would suggest that I would have a high amylase production.