Hydrolysis of Macromolecules Essay

4. There are other types of reagents used to determine what type of biomolecule a substance is. For example, copper ions present in Benedict’s reagent reacts with the free end of any reducing sugars, such as glucose, when heated. Originally blue in color, these copper ions are reduced by the sugar, and produce an orange-red colored precipitate. Alternatively, iodine-potassium iodide (IKI) may also be used when working with starch. IKI contains special tri-iodine ions which interact with the coiled structure of a starch

The use of multiple test tubes and Parafilm was used for each experiment. Catechol, potato juice, pH 7 phosphate buffer, and stock potato extract 1:1 will be used to conduct the following experiments: temperature effect on enzyme activity, the effect of pH on enzyme action, the effect of enzyme concentration, and the effect of substrate concentration on enzyme activity. For the temperature effect on enzyme activity, three test tube were filled with three ml of pH 7 phosphate buffer and each test tube was labels 1.5 degrees Celsius, 20 °C, and 60 °C. The first test tube was placed in an ice-water bath, the second test tube was left at room temperature, and the third test tube was placed in approximately 60°C of warm water. After filling the test tubes with three ml of the

0.0375 mg/ml Porcine Pancreatic Amylase Solution (amylase powder in 0.9% NaCl ), Iodine Solution; each solution were pipetted into each of the 5 test tubes with 5 ml of 1% starch. Each tube contained a 1% starch solution with a different pH. All tubes were at room temperature. Room temperature was 22C. 0.2 ml of porcine pancreatic amylase solution was then pipetted into each tube. A timer was started and every 3minutes the starch / amylase mixture were pipetted from each tube and pipetted into the spot plate for every sample tube, then the iodine solution were added to a spot plate cell for each sample. Iodine reacts with starch to change from yellow to deep blue /black in the presence of starch. A lightening of the blue/ black to a brown color will occur as less starch is present. Results were reported as (+) for presence of starch in the sample or (–) for the absence of starch. After every three minute increment had passed, these same

A cell, the building block of all living organisms, is composed of four fundamental biomolecules: proteins, carbohydrates, sugars and lipids. Proteins provide a vast amount of functions cells such as they serve as enzymes, provide structural support to cells, and act as antibodies. Reagents are used to spark a chemical reaction. The reagent used to detect protein traces in a substance is Biuret’s. Biuret’s will turn purple if proteins are present and blue if they are none. Biuret’s copper particles, have a charge of +2, are diminished to a charge of +1 when peptide bonds, which are in proteins, are present, creating the color change. Polysaccharides, which are carbohydrates, are most notably known to provide energy to the body, but they also help in breaking down fatty acids. Iodine is the reagent used to determine whether a substance has starch in it. The iodine/starch complex has energy levels that are only for retaining unmistakable light, giving the complex its extraordinarily dark black-blue shade. If there is no starch found, iodine will remain its natural yellowish-brownish color, but if starch is present, iodine will turn blue-black. Monosaccharides, which are sugars, like polysaccharides, provide the body with energy. To detect monosaccharides, the reagent, Benedict’s, is used. Benedict’s reagent is added to a test tube, then it is placed in

The purpose of this experiment is to exemplify how differences in molecular weight allow separation of polymers from their monomers. Methods of dialysis and gel filtration chromatography will be used to separate a glucose monomer from a starch polymer. Colorimetric glucose oxidase assay will be used to monitor the presence of glucose and a colorimetric iodine assay will be used to monitor the presence of starch in prepared solutions after separation

Introduction: I know prior to doing this experiment that iodine mixed with starch creates a dark color and that most objects, organic and inorganic, naturally experience isotonic reactions.

Moreover, CtXynGH30 also displayed activity against the polysaccharides having xylan main chain decorated with arabinose side chains such as arabinoxylans. Therefore a range of substrates showing the enzyme activities were treated with CtXynGH30 and the hydrolysed products were analyzed by TLC. The results showed that the enzyme is active against different polysaccharides and produces a series of oligosaccharides. The enzyme is active on xylan main chain polysaccharide substrates like beechwood-, birchwood- and 4-O-methyl glucurono-xylan and capable of releasing oligosaccharides such as xylose, xylobiose and other higher neutral and acidic oligosaccharides (Lane 1-3, Fig. 5). CtXynGH30 also acted over substrates having xylan main chain decorated with various degrees of arabinose side chains like oat spelt xylan, wheat arabinoxylan and rye arabinoxylan and producing xylobiose, xylotriose and other higher oligosaccharides (Lane 4-6, Fig. 5). Furthermore, the TLC profile of CtXynGH30 showed hydrolysis of arabinogalactan and more likely the release of arabino- oligosaccharides (Lane 7, Fig. 5), whereas, arabinan (sugar beet) and xyloglucan did not release any hydrolysed product (Lane 8-9, Fig. 5). The ability of CtXynGH30 to hydrolyse arabinoxylans apart from glucuronoxylans

In this lab experiment the action of the enzyme Amylase was observed on starch (the substrate). Amylase changed the starch into a simpler form, the sugar maltose, which is soluble in water. Maltose then breaks down the glucose chains of starch in the pancreas and intestines. Amylase is present in human saliva, and begins to act on the starch in the food while still in the mouth. Exposure to heat or extreme PH (acid or base) will denature proteins. Enzymes, including amylase, are proteins; if denatured enzymes can no longer act as a catalyst for the reaction. In the presence of potassium iodide, starch turns a dark purple color; however maltose does not react with I2KI. The rate of fading of starch allows a quantitative measurement of reaction rate.

A polysaccharide is when a numerous amounts of monosaccharides are joined together forming a molecule. Both glycogen and starch are polysaccharides. Glycogen is a energy storage carbohydrate found in animals. Glycogen in comparison to starch is much more highly branched. Starch is a polysaccharide that functions as a energy carbohydrate store in plants. This can be found mostly in potatoes and cereals. It is formed by the bonding together of many of glucose subunits into long chains. The actions of hydrolysis is the chemical breakdown of a compound due to the reaction of water. In order for hydrolysis to occur water must be added to the compound

enzymes that will be used during this lab to test the ability of amylase to break down starch ,a

In part II of the lab six small glass tubes were obtained in a test tube rack. Ten drops of distilled water were then added to test tube 1, five drops to tubes 2-4, and no drops in tubes 5 and 6. Five drops of 0.1M HCl were added to test tube 5 and five drops of 0.1M NaOH to test tube 6. Five drops of enzyme were then added to all tubes except tube 1. Tube 3 was then placed in the ice bucket and tube 4 was placed in the hot bucket at 80-900C for five minutes, the remaining tubes were left in the test tube rack. After the five minutes five drops of 1% starch was added to every tube and left to sit for ten minutes. After ten minutes five drops of DNSA were then added to all the tubes. All the tubes were then taken and placed in the

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

This experiment consisted of setting up a control group of starch in various temperature and then placing both fungal amylases and bacterial amylases in a mixture of starch and placing the solution of amylase and starch in various temperatures of water. After a certain amount of time- different amount of time needs to be used in order to have reliable results- iodine is added in a well on spot plates, then two drops of the mixture of amylase-starch is added from each temperature used, by adding iodine into the plates the mixture will show how much starch was hydrolyzed, this is used to calculate the amount of

For this experiment, we have to prepare our phosphorylase which extracted from a potato. We prepared by weighed about 250 grams of peeled potato and cut it into cubes. The extracts then blended with 100mL of 0.1M NaF. After filtered the contents into a clean 250mL centrifuge bottle, we centrifuged it for 3 minutes. Then, separated the supernatant into a centrifuge bottle, which is our phosphorylase preparation. The enzyme assay used in this experiment today is the iodine test. As the iodine reacts with starch, it will form a brown, blue or black precipitate due to the iodine ions forcing into a linear arrangement. The endpoint of the enzyme reaction indicates the presence of starch by using the iodine test to determine. The faster the endpoint is reached, the less active the phosphorylase is.

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).