...the process of hydrolysis occurring in different macromolecules by using the biochemical tests. This lab also indicates the hydrolysis of macromolecules when heat or acidity is applied to the molecules or compounds. The molecules that are being observed in the experiment are the polysaccharides and proteins, two main components that play an important role in cell biology. Hydrolysis could be done by applying heat, acid treatments, enzymic reaction, and bacteria involvement. IKI test and Benedict’s test is used in the experiment in order to detect the type of carbohydrate present in the testing tube containing the testing sample. In this experiment Hydrolysis is done through different methods. The first method is the hydrolysis of polysaccharides by adding acid or heat to the experiment. Starch is the polysaccharide being used in the experiment that is diluted with water. Starch solution, water, and glucose are tested for the presence of polysaccharides in it. The results are recorded in “before treatment” section. Same process is repeated for the benedict’s test that includes slight boiling, for the indication of reducing sugars. HCl is added to the test tubes that are placed into the bath tub at boiling temperature. The results are recorded in “after treatment” sections for both IKI and Benedict’s test. Color change is observed in the starch after adding heat. Results indicate that change in heat and acidity affect the hydrolysis. Glucose is a monomer while starch is a polysaccharide...
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...Experiment 8. The hydrolysis of starch with hydrochloric acid (a) Prepare a water bath by half filling a 250 cm3 beaker with warm water and heating it to boiling point on a tripod and gauze, with a Bunsen burner. When the water boils, reduce the flame to keep the water at boiling point. (b) Label four test-tubes 1-4. (c) Copy the table given below into your notebook. (d) In each tube place 5 cm3 3% starch solution. (e) Using a syringe or graduated pipette, add 3 cm3 Benedict's solution to the starch solution in tube 1 and place the tube in the boiling water bath for five minutes. (f) Rinse the syringe or pipette and use it to add 1 cm3 dilute hydrochloric acid to the starch solution in each of tubes 2, 3 and 4. Note the time and place all three tubes in the water bath. (They will be removed at five, ten and fifteen minutes respectively). (g) Remember to remove tube 1 from the water bath after five minutes if you have not already done so. (h) After five minutes, remove tube 2 from the water bath and cool it under the tap. Neutralize the acid by adding solid sodium bicarbonate, a little at a time, until the addition of one portion produces no fizzing. Place tube in the rack and return to tube 3. (i) After ten minutes in the water bath, remove tube 3, cool and neutralize the contents as described in (h). Place the tube in the rack. (j) After fifteen minutes in the water bath, remove tube 4; cool and neutralize as before, and place it in the rack...
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...shape of the molecule is extremely important to its function. Enzymes are composed of unique three-dimensional conformations, due to the complex folding during the secondary, tertiary, and quaternary, stages of protein production. Extreme pH levels, heat, concentration, and other factors can easily denature these exclusive structures. α-amylase is a biological catalyst found in the saliva of various organisms, including humans. It functions as a catalyst for the hydrolysis of starch products located in consumed foods. Chemically, starch is comprised of two different molecules, amylose and amylopectin. The glucose molecules in amylose are connected in a liner/straight manner, whereas, the glucose in amylopectin are arranged in a spiral shape. These unique linkages are what give this molecule its overall shape, and ultimately, its function. Starches produced in plants are normally a combination of both these molecules at a 30:70 ratio favoring amylopectin. The standard experiment done to detect if a substance contains starch is a reaction with Iodine (I2KI). The I2 in the reaction will react with the amylose molecules, changing its color from a yellow-amber color, to a dark purple color (Smith, 2000). In humans and countless other animals, α-amylase plays a vital role in the process of breaking down sugars and...
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...Practical 3 Investigation of Action of Saliva and Hydrochloric Acid in Two Carbohydrate Solution | Objective: 1. To show the action of saliva in two carbohydrate solutions. 2. To show the action of hydrochloric acid in two carbohydrate solutions. Apparatus & Equipment’s: Boiling tubes Metal test tube racks Beaker Graduated plastic dropper Water bath,~37°C Water bath,~95°C Stop watch Test tube holder Materials: Carbohydrate solution A Carbohydrate solution B Benedict’s solution 3M Hydrochloric acid 3M Sodium hydroxide Procedures: 1. Prepared two boiling tubes with containing 1 ml solution A and 1 ml solution B respectively. 1 ml Benedict’s solution was added to each tube and heated both tubes together in the (~95°C) water bath for two minutes. Then, recorded the results in table 1. 2. Added a few drops of fresh solution A and B separately spaced on a white tile. On each solution, added 1-2 drops of iodine solution and mixed with pen cover. Recorded your observations in the table 1. 3. Pipetted 2 ml solution B into each of four boiling tubes. The tubes were labelled 1, 2, 3 and 4 respectively near mouth of tube. Labelled your group name. 4. Placed tubes 1 and 2 in a water bath of ~37°C. 5. Salivated into a small beaker until it reached about 5 ml. 6. At the same time, step (6) and (7) was to be done approximately. Measured out 4 ml of the saliva prepared in step (4) and pipetted 2 ml each into tubes 1 and 4. The contents of the tubes shook well to ensure...
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...This is Google's cache of http://www.allsands.com/science/amylaseenzymeh_wpp_gn.htm. It is a snapshot of the page as it appeared on 9 Jul 2011 05:17:19 GMT. The current page could have changed in the meantime. Learn more Text-only version Home You Are At: AllSands Home > Science > Amylase enzyme: the effects of temperature About Contact Amylase Enzyme: The Effects Of Temperature Amylase is an important metabolic enzyme. Its function is to catalyze the hydrolysis of starch into glucose. At high temperatures, Amylase becomes denatured, denatured amylase no longer catalyzes the hydrolysis of starch into glucose. Amylase is an important metabolic enzyme. Its function is to catalyze the hydrolysis of starch into glucose. This particular enzyme, which is found in all m ammals, speeds up specific digestive processes which take place along the digestive track running from the mouth to the small intestines. Amylase's essential role in digestion makes it an attractive prospect for research. The human body must be kept within several degrees of 37° Celsius for biological functions to continue working. If body heat exceeds 37°C by too much cells become impaired or permanently damaged, at lower temperature metabolism decreases without permanent damage until ice crystals form in the cells. What happens to Amylase at extreme temperatures? Perhaps the answer to that question will give insight into metabolism's reaction to low temperatures. There are over 700 enzymes which have currently...
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...The purpose of this laboratory exercise was to perform tests necessary to be able to distinguish one microorganism from 10 others. Using a series of biochemical tests and characteristics, unknown #22 was concluded to be Pseudomonas aeruginosa. A dichotomous key was mapped out and used during this process. Using this provided guidance as well as organization as to what the result may be. Upon obtaining the unknown organism, it was important to make a streak plate of the bacteria on TSA. The purpose of doing so ensures that we have pure cultures of the unknown to be used in further testing and not a mixed culture. The first test used was a gram stain. It is a differential stain that helps distinguish between gram-positive and gram-negative bacteria. After performing the gram stain, it was clear that the unknown was a gram negative due to its pink color. Gram staining involves doing a simple smear, drying, and then heat fixing. Then using the staining technique with crystal violet, gram’s iodine, ethanol, and safranin, a pink or purple color should result when looking at the slide under a microscope. A gram-positive, or purple stained bacteria, means that there are multiple layers of peptidoglycan in the cell wall. A gram-negative means that there in a thin layer of peptidoglycan that is removed by the ethanol and stained pink by the counterstain. All gram-positive bacteria could now be ruled out. The gram-positive bacteria included: Enterococcus faecalis, Bacillus subtilis...
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...BIOLOGY UNIT 2 F212 Describe how hydrogen bonding occurs between water molecules, and relate this, and other properties of water, to the roles of water in living organisms (HSW1); A water molecule is made up of two hydrogen bonds and one of oxygen. The atom forms a triangular shape, although the molecule has no overall charge, the distribution of negatively charged electrons is uneven because the oxygen atom draws them away from the hydrogen. This consequently makes oxygen slightly negative while hydrogen has a slightly positive charge. We call this a dipolar molecule. Hydrogen bond Different poles attract , therefore the positive pole of one water molecule will attract the negative pole of another water molecule. The force of attraction between the opposite charges are known as hydrogen bonds. Although the bond is weak it forms important forces that gives water its properties. Properties of water * Specific heat capacity: because water molecules stick together it takes more energy to break them apart, for this point the boiling point of water is higher than expected. Water there for acts as a buffer against sudden temperature variations, making the aquatic environmental temperature stable * Cohesion and surface tension: the tendency of molecules sticking together is known as cohesion. With its hydrogen bonding water has large cohesive forces and these allow it to be pulled together through a tube such a xylem vessel in plants. * Density of water: water...
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...and function best under optimal conditions or temperatures related to the environment and the host organism where the enzyme is found. In this experiment, the digestive enzyme amylase was used in order to determine the optimal temperature for enzyme function from human and fungal (Aspergillus oryzae) sources in various temperatures. The digestive enzyme amylase that will be tested is responsible for the breakdown of the polysaccharide starch into the monosaccharide maltose. In addition, using Iodine as a test marker for the presence of starch, the ability of amylase to break down starch to maltose was also investigated. In the presence of starch, Iodine turns from yellow to black. To that end, human saliva, containing amylase, and fungal amylase were each added to separate test tubes along with a mild starch solution and placed at various temperatures as follows, 0C, 40C, 60C and 95C. The mixture of amylase and starch was then plated onto spot wells containing Iodine as a marker for starch which yielded a color reaction based on the amount of starch versus the break down product, maltose. The results showed that human amylase functions optimally at 40C as demonstrated by an abrupt color change from black to yellow signifying the quick breakdown of the amylase to maltose. Conversely, fungal amylase showed an abrupt color change at both 40C to 60C. The results analyzed correlate well with the expected in vivo environment of both organisms. The body temperature in humans is around...
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...polymerize when – A condensation reaction occurs – – Between two hydroxyl groups Resulting in a covalent bond called a glycosidic linkage Glycosidic Linkages § The glycosidic linkages can form – – § Between any two hydroxyl group The location and geometry of these bonds vary widely among polysaccharides α-1,4-glycosidic linkage and β-1,4- glycosidic linkage – Both linkages are between the C-1 and C-4 carbons Their geometry is different § – – α and β refer to the contrasting orientations of the C-1 hydroxyls They are on opposite sides of the plane of the glucose rings Watch this video https://www.youtube.com/watch?v=LBwSh8Q3sgs Types of Polysaccharides • Plants store sugar as starch – • • Mixture of branched (amylopectin) and unbranched (amylose) α-glucose polymer Animals store sugar as glycogen – Highly branched α-glucose polymer Cellulose: a...
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...factor on the activity of an enzyme Research Question: How does pH effect the activity of the enzyme amylase and the hydrolysis of starch? Hypothesis: Amylase is an enzyme that acts on starch. pH is capable of altering the structure of the active site centre in the enzyme leading to denaturation. At each pH, the enzyme activity would be relatively different. Ideally the optimum pH is 6.0 when the enzyme works best and the fastest, however if the pH is higher or lower the hydrolysis of starch will be slower. Changing the pH above the optimum will sometimes lead to denaturation of the enzyme and the change in shape of the active site. (http://www.nuffieldfoundation.org/practical-biology/investigating-effect-ph-amylase-activity) Variables: Independent: The pH of the buffer solution. Dependent: The time taken for the solution to turn an opaque (lighter) blue from dark blue. Controlled: The volume of iodine solution; the volume of starch solution; the volume of buffer solutions; the volume of amylase solution; the temperature at 40°c; the time it is left in the water bath; and the concentration of the solutions. I will be using a water bath to keep the temperature constant, and to manipulate my independent variable, I will be using pH 4, 5, 7, 9, 10 by adding different pH buffer solutions with the amylase. * 50ml Amylase solution * 100ml Starch solution * 100ml Buffer solutions with pH 4, 5, 7, 9, 10 * 20ml Iodine solution Apparatus: * Test tubes...
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...IDENTIFICATION OF UNKNOWN BACTERIA It is virtually impossible to identify bacteria based on physical characteristics alone. This is due to the fact that there are only a few basic shapes and physical features commonly seen in the prokaryotic world. Instead, biochemical testing has been used to make bacterial identification down to the “species” level. These schemes are based on creating and matching biochemical profiles of the production of enzymes, acids and gases by isolated pure cultures of a given microorganism. Identification schemes and flow charts can be found in reference texts such as “Bergey’s Manual of Determinative Bacteriology” or “The Prokaryotes”. Each group of students will receive a TSA slant or broth containing a pure culture of an unknown bacterium belonging to the Family Enterobacteriaceae. It is the responsibility of the group to maintain stock cultures of the organism provided. Working stock cultures will be used to inoculate the various biochemical test media over the next several weeks and should be fresh and free from contaminants. A reserve stock culture should be made and after incubation and comparison with the original slant, kept with the original slant in the refrigerator. It is critically important that aseptic techniques are used during transfers and inoculations to prevent contamination of your cultures. If contamination is suspected, you will be able to fall back to your reserve stock. If you fail to maintain a reserve stock...
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...dark red plate used to distinguish gram negative, enteric, lactose fermenter bacteria from others. The starch plate was a clear plate used to test for hydrolysis. Hydrolysis is the breakdown of a compound with water. A positive for this test resulted in a clearing area around the bacteria after the addition of iodine called the “Zone of Hydrolysis.” The Casein plate was a skim plate used to test for the “Zone of Proteolysis.” The “Zone of Proteolysis” is a white clearing due to the breakdown of the Casein milk plate. The Oxidase plate was used to do the oxidase test. The Lipid Hydrolysis plate was...
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... solutions. Results: Table 1: Observations made when two carbohydrate solutions provided in laboratory were tested with Benedict and Iodine solution. | Observations | Conclusions | Solution A | Benedict’s test: An initial blue translucent mixture turned to brick-red opaque solution and moderate amount of precipitate settled after heated at a high temperature for two minute. | Presence of reducing sugar | | Iodine test: The translucent colouration of the mixture retained its yellowish-brown colour. | Absence of starch | Solution B | Benedict’s test: The translucent colouration of the mixture remained its blue colour. | Absence of reducing sugar | | Iodine test: The initial yellowish-brown translucent mixture turned to bluish-black opaque solution when solution was mixed. | Presence of starch | Table 2: Colour reactions of Benedict’s test for saliva and 3M hydrochloric acid in two carbohydrate solutions provided in laboratory. Tube | Contents | Temperature (ºC) | Benedict’s Test—Colour...
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...The purpose of this experiment is to examine the effect that enzyme concentration has on reaction time and the effect that substrate concentration has on enzyme reaction. Enzymes are biological catalysts that catalyze different chemical reactions. In general, enzymes are proteins and they are each specific to specific chemical reaction. In order for enzymes to process properly, they should maintain a specific three dimensional structure. When enzymes function, they combine with their substrates (reactant) to form susbtrate-enzyme complex. Then this complex converts into a product and unaltered enzyme. Substrate + Enzyme Substrate-Enzyme Complex Product + Enzyme OR Substrate –Enzyme Product (From this equation, in general, the reaction of enzyme is irreversible.) Some of the factors that affect the rate of reaction are temperature, pH, enzyme concentration, substrate concentration, product concentration, etc. The rate of reaction is affected by the level of pH. The extreme level of pH can denature enzyme and result loss of its action. The optimum pH is 14 and this is the level of pH where the rate of reaction is the highest. Temperature also affects the rate of reaction. As temperature increases, the rate of reaction increases as well; however, it increases until the optimum temperature. After optimum temperature, the enzyme denatured. The concentration of enzyme and substrate affect the rate of reaction. In theory, the higher the concentration of substrate, the...
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...Objective: To identify the components of the two solutions with Iodine test and ... Discussion: ... Why You Add Hydrochloric Acid in Hydrolysis of Starch. Ingestion of saliva during carbohydrate feeding by ... - SciELO www.scielo.br/scielo.php?pid=S0074-02762006000100016&script... by RR Cavalcante - 2006 - Cited by 5 - Related articles Saliva ingestion by phlebotomine during the carbohydrate ingestion phase is ... The presence of saliva in each type of solution or substrate offered, as well as ... 0.2 ml of apyrase assay buffer pH 8 (50 mM Tris/HCl buffer containing 1 mM CaCl2, ... Saliva ingestion occurred under each condition investigated, as indicated by ... Experiment: investigation of action of saliva and hydrochloric ... https://answers.yahoo.com/question/index?qid... Jun 25, 2012 - 1) Name of enzyme involved 2)specific action(s) of enzymes involved. Two hydrolytic enzymes and an epistemological–historical ... www.scienceinschool.org/2007/issue4/enzymes Science in School Sep 3, 2007 - 5 M sodium hydroxide (NaOH); 5 M hydrochloric acid (HCl); Saliva. ... To demonstrate the test methods, test all four carbohydrate solutions ... Discussion ... more concentrated than is necessary for the activity and that using a ... The authors have recommended the Fehling’s test for this investigation. What is the objectives of investigation of action of saliva and ... www.answers.com › Wiki Answers › Categories › Science › Biology ... of investigation of action of saliva...
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