Investigation the effect of the temperature on the activity of amylase
Aim:
The aim of the experiment is to detect how different ranges of temperatures as a significant factor on the rate of the reaction has impact on the prosperities of enzyme amylase and carbohydrate as starch under independent variable which is time required for reaction and dependant variable which is rate of the reaction.
Introduction:
Enzymes are known as globular proteins which increase the rate of the specific biochemical reaction under the specific circumstances the reaction can occur. All enzymes have active site where the reaction is catalysed. Active site of the enzyme is a place where enzyme has the right shape and functional group to bind in a specific way the particular and precise substrate molecule. It is determined by the enzyme’s structure, which is complex of foiled and coiled in 3-dimentional globular (spherical) protein with functional group like carboxyl group (COOH), amino group (NH2 ) and R group to specify uniquely about the prosperities like hydrophobicity, hydrophilicity, weak acid or weak base so the reaction can take place. Almost all enzymes have just one kind of substrate molecule on which they can act. Amylase is an enzyme found in salivary glands named ptyalin and the pancreas also secretes the enzyme. Amylase act as a catalyst in breaking down the starch’s long chain glucose unit joined together by glycosidic bonds (covalent bond that joins a carbohydrates molecule to
There are many types of enzymes and each has a specific job. Enzymes are particular types of proteins that help to speed up some reactions, such as reactants going to products. One of them is the amylase enzyme. Amylases are found in saliva, and pancreatic secretions of the small intestine. The function of amylase is to break down big molecules of starch into small molecules like glucose; this process is called hydrolysis. Enzymes are very specific; for example, amylase is the only enzyme that will break down starch. It is similar to the theory of the lock
Enzymes are biological catalysts that speed up chemical reactions, without being used up or changed. Catalase is a globular protein molecule that is found in all living cells. A globular protein is a protein with its molecules curled up into a 'ball' shape. All enzymes have an active site. This is where another molecule(s) can bind with the enzyme. This molecule is known as the substrate. When the substrate binds with the enzyme, a product is produced. Enzymes are specific to their substrate, because the shape of their active site will only fit the shape of their substrate. It is said that the substrate is complimentary to their substrate.
As the temperature increases, so will the rate of enzyme reaction. However, as the temperature exceeds the optimum the rate of reaction will decrease.
The purpose of this experiment was to determine (1) the reaction rate of an amylase enzyme in starch and (2) the environmental factors that can affect the enzymatic activity. The hypothesis, in relation to the enzymatic activity by variables such as the substrate concentrations, temperature, PH and chemical interactions on the rate of reaction, stated
amylase enzyme and the optimal temperature for fungal and bacterial amylase. In order to make
During these experimental procedures, the implication of multiple different temperatures on fungal and bacterial amylase was studied. In order to conduct this experiment, there were four different temperatures used. The four temperatures used were the following: 0 degrees Celsius, 25 degrees Celsius, 55 degrees Celsius, and 80 degrees Celsius - Each temperature for one fungal and one bacterial amylase. Drops of iodine were then placed in order to measure the effectiveness of the enzyme. This method is produced as the starch test. The enzyme was tested over the course of ten minutes to determine if starch hydrolysis stemmed. An effective enzyme would indicate a color variation between blue/black to a more yellowish color towards the end of the time intervals, whereas a not so effective enzyme would produce little to no change in color variation. According to the experiment, both the fungal amylase and bacterial amylase exhibited a optimal temperature. This was discovered by observing during which temperature and time period produced a yellow-like color the quickest. Amylase shared a similar optimal temperature of 55 degrees Celsius. Most of the amylases underwent changes at different points, but some enzymes displayed no effectiveness at all. Both amylases displayed this inactivity at 0 degrees Celsius. At 80 Celsius both the enzymes became denatured due to the high temperatures. In culmination, both fungal and bacterial amylase presented a array of change during it’s
“Enzymes are proteins that have catalytic functions” [1], “that speed up or slow down reactions”[2], “indispensable to maintenance and activity of life”[1]. They are each very specific, and will only work when a particular substrate fits in their active site. An active site is “a region on the surface of an enzyme where the substrate binds, and where the reaction occurs”[2].
Effect of varying Temperatures on Enzymatic Activity of Bacterial and Fungal Amylase and hydrolysis of Starch
Enzymes are catalysts that affect the rate of a reaction without itself being affected or altered. In this experiment, the enzyme being experimented with is amylase. This enzyme can be found in human saliva and helps to hydrolyze starch into maltose. There are three different parts to this experiment; concentration, pH, and temperature. Each is to test the presence of amylase and the different affects it has on certain substances. Concentration is to determine what effect the variation of will have on the rate of the reaction, pH to determine the optimum pH for the activity of amylase, temperature is manipulated to see how amylase would react in different degrees of temperature and how active it is.
Most enzymes are made of the macromolecule proteins. The job of an enzyme is to speed up the breakdown of chemicals and reduce the activation energy needed to do so. Enzymes are proteins folded into complex shapes that allow smaller molecules to fit into them. The place where these substrate molecules fit is called the active site. The enzyme then either breaks down the protein or creates a new larger protein. The enzyme amylase is found in saliva and breaks down the substrate starch,or carbohydrates. Amylase is produced in the salivary glands, pancreas and small intestine. Temperature and pH levels affect the reactivity of the enzyme. Changes in an enzyme's environment cause the enzyme to become denatured or misshapen causing the substrate
Enzymes are proteins that act as catalysts and help reactions take place. In short, enzymes reduce the energy needed for a reaction to take place, permitting a reaction to take place more easily. Some enzymes are shape specific and reduce the energy for certain reactions. Enzymes have unique folds of the amino acid chain which result in specifically shaped active sites (Frankova Fry 2013). When substrates fit in the active site of an enzyme, then it is able to catalyze the reaction. Enzyme activity is affected by the concentrations of the enzymes and substrate present (Worthington 2010). As the incidence of enzyme increases, the rate of reaction increases. Additionally, as the incidence of substrate increases so does the rate of reaction.
In this experiment we wanted to determine the optimal temperatures for fungal, Aspergillus oryzae, and bacterial, Bacillus licheniformis. In order to see if any of the starch was broken down, Iodine was mixed with the starch-amylase substance. In four spot plates, the groups labeled the different temperatures, once the iodine came in contact with the starch, the result would be a reaction that turns the fluid into this dark blue/black color. In a span of 10 minutes, with occasional check ups on the solution every 2 minutes, the amylase-starch solution was placed into five types of temperature, all being Celsius. The five temperatures were 0 degrees, 25 degrees, 55 degrees, and 85 degrees Celsius. The solution would change colors, so in order to measure the changes, a scale was used. Such scale was a 1-5 scale, with colors next to each number. One being the lightest color, or yellow, and 5 being the darkest color, or black. Based on the change of color, we could tell how fast it hydrolyzed the starch in a span of 10 minutes. To keep record of the results, the results were put in Data Tables used from the Lab Manual. The average optimal temperature for Bacteria Amylase was 85 degrees Celsius, while the Fungal was 55 degrees Celsius. You can see this by looking for the
In this lab our group observed the role of pancreatic amylase in the digestion of starch and the optimum temperature and pH that affects this enzyme. Enzymes are located inside of cells that increase the rate of a chemical reaction (Cooper, 2000). Most enzymes function in a narrow range of pH between 5 through 9 (Won-Park, Zipp, 2000). The temperature for which enzymes can function is limited as well ranging from 0 degrees Celsius (melting point) to 100 degrees Celsius (boiling point)(Won-Park, Zipp, 2000). When the temperature varies in range it can affect the enzyme either by affecting the constant of the reaction rate or by thermal denturization of the particular enzyme (Won-Park, Zipp, 2000). In this lab in particular the enzyme, which was of concern, was pancreatic amylase. This type of amylase comes from and is secreted from the pancreas to digest starch to break it down into a more simple form called maltose. Maltose is a disaccharide composed of two monosaccharides of glucose. The presence of glucose in our experiment can be identified by Benedicts solution, which shows that the reducing of sugars has taken place. If positive the solution will turn into a murky reddish color, where if it is negative it will stay clear in our reaction. We can also test if no reduction of sugars takes place by an iodine test. If starch is present the test will show a dark black color (Ophardt, 2003).
The Effects of Enzyme Concentration on the Activity of Amylase To investigate the effect of Amylase concentration on its activity. the relative activity of Amylase is found by noting the time taken for the starch substrate to be broken down, that is, when it is no longer gives a blue-black colour when tested with iodine solution. This time is referred to as the achromatic point. Equipment: v Amylase solution 0.1% v Starch Solution 1.0% v Distilled water v Iodine in potassium iodine solution v White tile and polythene pipette v Graduate pipettes or syringes v Test tubes in rack v Beaker (used as water bath) v Stopwatch, Thermometer v Eye Protection
The enzymes inside the human body is a protein molecule which is a biological catalyst containing 3 types of characteristics. The first characteristic of an enzyme is to increase inside the human body depending on the rate of reaction. Secondly most of the enzymes inside the human body will take control and only act specifically upon one reactant, this is called the substrate, and this is the enzyme that produces products. Third most important is the enzymes strongly regulate in a circle, from a low concentration (activity) to a high concentration (activity) then back around vice versa. The enzymes can strongly be affected in the changes of both the temperature and the different ph levels. Every individual enzyme works to a certain extent reacting to a certain ph level and temperature, its activity decreasing at values above and below that point due to denaturation. For enzymes, denaturation can be defined as the loss of enough structure rendering the enzyme inactive. The tertiary structure of the enzymes and their function also includes the non covalent force in creating and determining the shape and the structure of all enzymes. The salivary amylase is found in the salivary glands inside the human body which is produced by the ptyalin. Maltose and Isomaltose are a