...BIOCHEMISTRYLABORATORY(Midterm)Geromil J. Lara, RMT, MSMT 2. ACTIVITY 1A /1B Subcellular Components of the Living Cell• 5% Trichloroacetic Acid – Suspending medium – Able to precipitate proteins – Good fixative and preservative 3. ACTIVITY 1A /1B Subcellular components of the Living Cell• Sediment 1 – Nuclear Fraction – Nuclei and Unbroken Cells• Sediment 2 – Mitochondria• Sediment 3 – Microsome – Proteins, Enzymes, Inorganic Ions 4. ACTIVITY 1A /1B Subcellular components of the Living Cell• Qualitative Tests – Carbohydrates • Molisch Test • Benedict’s Test – Proteins • Biuret Test • Xanthoproteic Test – Lipids • Sudan Test • Acrolein Test 5. ACTIVITY 1A /1B Subcellular Components of the Living Cell• Carbohydrates – Molisch Test 5 drops of the supernate + Molisch Reagent shake Layer with 1 mL conc. H2SO4Control: 1% LecithinMolisch Reagent: α-Naphthol dissolved in Ethanol 6. ACTIVITY 1A Subcellular components of the Living Cell• Carbohydrates – Molisch Test 5 drops of the supernate + Molisch Reagent shake Layer with 1 mL conc. H2SO4Principle: when sugar solution is mixed with alpha- naphthol is brought in contact with conc. H2SO4, a violet ring is formed at the junction of the 2 liquids 7. ACTIVITY 1A Subcellular components of the Living Cell• Carbohydrates – Molisch Test 5 drops of the supernate + Molisch Reagent shake Layer with 1 mL conc. H2SO4H2SO4 : acts as dehydrating agent forming furfural derivatives which interact with alpha-naphthol liberating a colored compound 8. ACTIVITY 1A /1B Subcellular Components...
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...Introduction & Hypothesis: Enzymes are important for every living organism, because they are the reason that reactions occur. Although most reactions would take place without enzymes, enzymes allow these chemical reactions to happen at a much faster rate, therefore making cells more efficient (Reece, 2011). Enzymes are catalysts, and almost always proteins, that speed up the rates of reactions by lowering the activation energy without being consumed in the reaction (Helms, 1998). Throughout this experiment, four procedures will be performed to indicate the factors that alter the functioning of enzymes, and the importance that these factors are to be in correct levels in order for a cell to function properly. Proteins are macromolecules with unique polypeptide chains that make up their structure (Reece, 2011). The functions of proteins are dependent on their structure, and since enzymes are almost always proteins, the structure of the enzyme is very important for the enzyme to function. However, there are factors that can disrupt the structures of enzymes. These can be environmental factors such as temperature and pH, or they can be concentration changes, such as an increase or decrease in enzyme or substrate concentration (Eed, 2013). Temperature is an environmental factor that can alter enzyme activity (Reece, 2011). An increase in the kinetic energy of a solution results in an increase in temperature (Reece, 2011). As the temperature increases, the molecules in the...
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...Temperature can affect a lot of different factors hence its effect on enzyme activity is very complex. It affects the speeds of molecules, the activation energy of the catalytic reaction and the thermal stability of the enzyme and substrate. At low temperatures (say at around 0 centigrade) the rate of enzyme reaction is very slow. The molecules have low kinetic energy and collisions between them are less frequent and even if they do collide the molecules do not posses the minimum activation energy required for the reaction to occur. It can be said that the enzymes are deactivated at low temperatures. An increase in temperature increases the enzyme activity since the molecules now possess greater kinetic energy. The rate of enzyme activity is highest between 0-40 centigrade and this increase is almost linear. After 40 the rate of reaction starts to decrease. This is because the increase in temperature after 40 does not increase the kinetic energy of the enzyme but instead disrupts the forces maintaining the shape of the molecule. The enzyme molecules are gradually denatured causing the shape of the active site to change. Temperatures above 65 centigrade completely denature the enzymes. There are some enzymes known as ‘extremophiles’ found in thermophillic organisms. They retain activity at 80 centigrade. Test each separate temperature more than twice: This is to prove that each temperature is affecting the enzymes the same way each time and that the previous result...
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...Each group was successful in gathering results for the different factors and it proves just how much the peroxidase became affected within the testing environments. In terms of the enzyme concentration, both of the different conditions (double and half) resulted with the absorption readings at 470 nm to steadily increase over the course of 5 minutes. This result was caused because there were more enzymes present to react with the two substrates, hydrogen peroxide and guaiacol, therefore affecting the overall enzyme activity to become higher. Moving onto the temperature group, the results were a bit staggered. Over the course of the 5 minutes, the cold, body, and warm test tube results showed an increasement in the absorption readings, while...
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...In lab 7, Factors affecting Enzyme Activity, there are two parts; Effects of concentration on enzyme activity and effects of temperature on enzyme activity. In the first lab with concentration the materials needed to preform this experiment will be a LabQuest, Vernier O2 Gas Sensor, Nalgene bottle, enzyme suspension, 3.0% H2O2 , stop watch, goggles, three test tubes and dH2O and a 10 ml graduated cylinder.The first step in the procedure is to obtain three test tubes and label them 1, 2, and 3. With a graduated cylinder, fill the three tubes with 5 ml of 3.0% H2O2 and 5 ml of water. Add five drops of enzyme suspension to test tube labeled 1. Wait thirty seconds and pour solution into a Nalgene bottle. Place the O2 gas sensor into the bottle...
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...Chapter 3 Enzymes Learning Outcomes Candidates should be able to: (a) explain that enzymes are globular proteins that catalyse metabolic reactions; (b) explain the mode of action of enzymes in terms of an active site, enzyme/substrate complex, lowering of activation energy and enzyme specificity; (c) [PA] follow the progress of an enzyme-catalysed reaction by measuring rates of formation of products (for example, using catalase) or rates of disappearance of substrate (for example, using amylase); (d) [PA] investigate and explain the effects of temperature, pH, enzyme concentration and substrate concentration on the rate of enzymecatalysed reactions; (e) explain the effects of competitive and non-competitive inhibitors on the rate of enzyme activity; (f) use the knowledge gained in this section in new situations or to solve related problems. Enzymes are globular protein which act as catalysts • Enzymes are protein molecules defined as biological catalysts which speed up a chemical reaction and remain unchanged at the end of reaction. • Enzyme names end in –ase E.g. amylase, ATPase • Enzyme are globular proteins. Enzymes are globular protein which act as catalysts • Enzyme molecules are coiled into a precise threedimensional shape, which hydrophilic R groups (side-chains) on the outside of the molecule ensuring that they are soluble. Enzymes are globular protein which act as catalysts • Enzyme molecules have active site. • The active site of an enzyme is a region...
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...Introduction Enzymes are protein molecules that catalyze chemical reactions in all living organisms. Enzymes allow living organisms to carry out complex chemical activities at low temperatures, but can’t cause a reaction that hasn’t occurred in their absence. Also, enzymes are thought to speed up reactions by bringing reacting molecules together to increase the chances that a reaction will occur (Worthington Biomedical Corporation, 2015). Each enzyme has a specific active site where the substrates attach. Many factors can affect enzyme activity such as temperature, pH, and the presence of inhibitors (John W. Kimball, 2014). The purpose of this lab was to examine factors affecting the enzyme function of peroxidase. In the 19th century French chemist Louis Jacques discovered catalysts. Catalysts are substances that enable a chemical reaction without participating in it, which led to specifically peroxidases. The structure of peroxidase is a very large enzymatic protein, and has complex molecules with complicated shapes involving multiple folding’s. The activity of peroxidase is dependent on pH. It exhibits maximum activity at a pH between 6.5 and 7.0. The activity of the enzyme is reduced when pH levels are increased. Peroxidase promotes the oxidation of various compounds naturally of peroxides, where hydrogen peroxide is reduced to form water (Wikimedia Foundation, 2015). Also peroxidases break compounds down into harmless substances by adding donor molecules. During this lab...
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...OF ENZYMES Enzymes are extremely important and without them, the reactions in living organisms would be so slow they would hardly proceed at all. They enable metabolic reactions to proceed rapidly at low temperatures, and as well as speeding up reactions they also control them. There are two main groups of enzymes: intracellular and extracellular. INTRACELLULAR: Occur inside cells where they speed up and control metabolism. EXTRACELLULAR: Produced by cells but achieve their effects outside the cell – includes digestive enzymes that break down food in the gut. Each enzyme is usually specific to particular reactions, and are pH sensitive, with every enzyme having its own range of pH in which it functions best. They are not destroyed by the reactions which they catalyse, meaning they can repeat a reaction over and over. (McMonagle,2015) In an enzyme controlled reaction, the substrate molecules bind with the enzyme to form an enzyme-substrate complex. The reaction then takes place and the product leaves the enzyme. As mentioned above, the enzyme (unchanged by reaction) can then be used again. Below is the equation, with the double arrow meaning that the reaction can go either way, depending on the amount of substrates and products – this ensures a equilibrium, meaning the enzyme will switch if there is an abundance of product and not enough substrates, and vice versa. (Advanced Biology, pg124, 2000) ENZYME + SUBSTRATE ENZYME-SUBSTRATE...
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...Ybor City Campus 1025C Laboratory Exercise 3: Characteristics of Enzymes Introduction What are Enzymes? Enzymes are very large and complex proteins that that function as catalysts. They are synthesized by the cell to perform very specific functions. Each type of enzyme has a very specific shape (conformation) which provides its specific function. The shape of an enzyme molecule is determined and maintained by many weak intermolecular interactions between many different parts of the molecule. A catalyst is a material that donates energy to reactants in order to reduce their activation energy. In other words, they allow reactions to occur with less energy input (i.e. more quickly or at lower temperatures). The very specific shape of enzymes allows them to “fit” into a reaction and reduce the energy necessary for the reaction to occur. If their shape is changed, their ability to function as a catalyst is reduced or eliminated. As biological catalysts, enzymes are important because they speed up the rate of the reaction they catalyze that would otherwise be too slow to support life. Factors Affecting Enzyme Activity The rate at which an enzyme works is influenced by several factors including temperature and pH. Enzymes are most effective as catalysts under optimum physical and chemical conditions; as conditions change away from optimum, enzyme activity decreases (Figure 1.). Changes in various environmental factors, such as temperature or pH, may affect proteins by altering their...
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...Enzyme Catalysis: Lab Report Introduction: In the lab there were 4 parts (A, B, C, D). Each part having a different reaction among it, an example of this reaction could be lab A when the catalysis was tested for activity. The reaction was to support the affect catalysis had on the hydrogen peroxide. This was a fizzing and bubbling state supporting a reaction was occurring. So one can infer that if enzyme catalysis brakes down hydrogen peroxide, then an abandons of catalysis would break down more hydrogen peroxide upon a quicker rate. Materials and methods: Part A: hydrogen peroxide, catalysis. After taking 10ml of H2O2 and mixing it with 1ml of catalysis for 30 seconds. After the reaction the data was recorded. Part B: H2H2, H4SO4, KMnO4 H2O. In the reaction an observation of 10ml of H2O2 in a 60ml container, next 1ml of distilled water is transferred, then 10ml H2SO4 is moved to a cup labeled baseline, finally 5ml of the solution is moved from baseline to titration and is titrated. Lastly the data is recorded. Part C: H2O2 (overnight), H2SO4, KMnO4. Basically the same reaction that occurred in part B was duplicated, but with an overnight solution of H2O2. It was in a baseline and the transferred to titration and then data was recorded. Part D: H2O2, H2SO4, KMnO4. With 60ml cups labeled 10sec, 30sec, 60sec,120sec, and 180sec catalysis was put to a decomposes test. 1ml of catalysis was transferred to a cup and then was swirled. Finally H2SO4 was added to stop the enzyme...
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...The Effects of Peroxidase on Enzyme Activity Gianna Crowe Bio Lab 117 October 16th, 2014 Most enzymes are proteins that speed up reactions and are characterized as catalysts. Enzymes work in such a way that when the right chemicals of a molecule are present for the enzyme, it will fully fit the shape. The part of the particular shape is called the active site of the enzyme, since this is where the reaction occurs. The molecule that the enzyme works on is called the substrate. An enzyme reaction includes a substrate (substance) that is converted to another product. The unique shape of the active site of the enzyme allows it to bind with only certain kinds of molecules, which is the substrate of the enzyme (Strobl 2014). A substrate binds to the active site of the enzyme to form a enzyme-substrate complex for a very short time, this then becomes part of a new formation and a new product of a specific reaction is formed then released freeing the active site, allowing the enzyme to repeatedly bind another substrate. Enzymes are produced by all living things, and are a necessity to life. They are responsible for constructing, synthesizing, carrying, dispensing, delivering, and eliminating the many chemicals associated in living organisms (Colpa 2014). An example for how enzymes work in living organisms would be the process of food digestion, enzymes work to break down food and speed up the digestion process. Factors that affect enzyme activity deal with environmental conditions...
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... Endo membrane structures: Endoplasmic reticulum, Golgi apparatus, Lysosome, peroxysome and vacuole. Nucleus-membrane-bounded organelle within a eukaryotic cell that contains chromosomes and controls the structure and function of the cell. Ribosome- Site of protein synthesis in a cell; composed of proteins and ribosomal RNA. Endoplasmic reticulum – System of membranous saccules and channels in the cytoplasm, often with attached ribosomes. Golgi apparatus – organelle consisting of sacs and vesicles that processes, packages, and distributes molecules about or from the cell Lysosomes- are membrane bounded vesicles produced by the golgi apparatus. Peroxisomes- a membrane bound vesicles that enclose enzymes. Vacuoles- membrane bround sacs 5. Energy related organelles: Mitochondria and Chloroplast. Basic idea about Photosynthesis. 6. The Cytoskeleton: names of different structures and their brief functions. Chapter 5 1. Membrane Structure and Function: The Phospholipid bilayer (Fluid mosaic model). 2. Different types of membrane proteins and their functions. 3. Permeability of the Plasma Membrane. It is selectively permeable. Transport through it. 4....
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...ioChapter 6 Metabolism: Energy and Enzymes The nature of energy and the laws of thermodynamics are discussed, followed by a detailed description of energy transformations that occur within the cell. The chemistry and functions of ATP are described. The role of enzymes in metabolism, oxidation-reduction reactions, and the cellular organelles in which these reactions take place are detailed. Chapter Outline 6.1 Cells and the Flow of Energy A. Forms of Energy 1. Energy is capacity to do work; cells continually use energy to develop, grow, repair, reproduce, etc. 2. Kinetic energy is energy of motion; all moving objects have kinetic energy. 3. Potential energy is stored energy. 4. Food is chemical energy; it contains potential energy. 5. Chemical energy can be converted into mechanical energy, e.g., muscle movement. B. Two Laws of Thermodynamics 1. First law of thermodynamics (also called the law of conservation of energy) a. Energy cannot be created or destroyed, but it can be changed from one form to another. b. In an ecosystem, solar energy is converted to chemical energy by the process of photosynthesis; some of the chemical energy in the plant is converted to chemical energy in an animal, which in turn can become mechanical energy or heat loss. c. Neither the plant nor the animal create energy, they convert it from one form to another. d. Likewise, energy is not destroyed; some becomes heat that dissipates into the environment. ...
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...arteries * result of atherosclerosis- plaque buildup in the coronary arteries * leads to blockages in the coronary arteries * heart becomes starved of oxygen and the vital nutrients * this leads to ischemia- restriction in blood supply to tissues, causing a shortage of oxygen and glucose needed for cellular metabolism * is the No. 1 killer in America * affecting more than 13 million Americans * risk for heart attack (Myocardial infarction-MI) and also stroke Myocardial Infarction (MI)/ Heart Attack Myocardial infarction (MI) (heart attack) is the irreversible necrosis (death) of heart muscle secondary to prolonged ischemia. CAUSES: * plaque from atherosclerosis and blood clot that blocks blood from flowing to the heart. This is the most common cause of heart attacks. * cocaine. The cause of heart attacks is not always known. Heart attacks may occur: * during resting or sleeping * After a sudden increase in physical activity * When active outside in cold weather * After sudden, severe emotional or physical stress, including an illness RISK FACTORS * age. Risk of heart disease increases with age. * gender. Men have a higher risk of getting heart disease than women who are still menstruating. After menopause, the risk for women gets closer to the risk for men. * genes or race. If parents had heart disease, higher risk. African-Americans, * smoking. * high cholesterol * high blood pressure . ...
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...* Key Topics: * Explain and give examples of why proteins are essential to cell function * The basic structure of an amino acid * Describe the 4 levels of protein structure, and give examples of each * Explain what an enzyme is, understand why enzymes are needed to help chemical reactions, and know the role of the active site * Revisiting the Theory of Chemical Evolution * Modern life arose through a series of endergonic chemical reactions. 1. Production of small organic compounds * i.e., formaldehyde (H2CO), hydrogen cyanide (HCN) 2. Formation of mid-sized molecules from these small compounds * i.e., amino acids, simple sugars 3. Mid-sized building blocks combine to form large molecules. * i.e., proteins, complex carbohydrates 4. Life became possible when one of these large molecules self-replicated. * Organic Molecules * Large organic molecules are called macromolecules * Four major categories of macromolecules: 1. Proteins 2. Nucleic Acids 3. Carbohydrates 4. Lipids * Several of these are long chains of smaller subunits. * The smaller subunits are known as monomers * The long chains of monomers are known as polymers * Four groups of Macromolecules * Building Macromolecules – Monomers and Polymers * Molecules, such as amino acids, are individual units called monomers. They link together (polymerize) to form polymers, such as proteins...
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