Unit 3 - Enzymes 3.1 – Enzyme Action, 3.2 – Effects of temperature and pH. SUFEATIN SURHAN BIOLOGY 5090 2012 SYLLABUS CHECKLIST Candidates should be able to: a) define enzymes as proteins that function as biological catalysts; b) explain enzyme action in terms of the ‘lock and key’ hypothesis; c) investigate activity. and describe the effect of temperature and pH on enzyme Enzymes and us Enzymes play a big role in our lives. When we eat a piece of bread, enzymes in our digestive system work to break down the complex carbohydrates found in the bread into simpler sugars for absorption into the blood stream. Digestive enzymes are also used in washing powders. They help to break down and remove stains caused by organic matter such as sweat, blood, curry, etc. What are enzymes? DEFINITION: Enzymes are biological catalysts made up of proteins which speed up the chemical reactions within an organism without themselves being chemically changed at the end of the process. Catalysts are chemicals that can speed up the rate of a reaction. There are two types of catalysts i.e. organic (biological) and inorganic catalysts. The substance acted upon by enzymes in an enzyme-catalysed reaction is called substrate. substance(s) produced in the reaction are known as product(s). The General Characteristics of enzymes 1. Enzymes are all proteins. 2. Enzymes are biological catalysts. 3. Enzymes reactions are reversible. 4. Each enzyme works best at a particular temperature and pH. 5. Enzymes are specific – they work on one kind of substrate only. 6. Only small amounts of enzymes are needed to do their job. 7. All enzymes are water soluble. 8. Rate of enzyme activity is affected by temperature and pH. Intracellular and Extracellular enzymes Intracellular enzymes are produced in living cell and work inside the same cell. They are found in the nucleus, cytoplasm, mitochondrion and chloroplasts. For example, catalase which breaks down harmful hydrogen peroxide in the liver cells. Extracellular enzymes are produced inside cells but are released from the cells to carry out their job. For example, amylase is produced in the salivary gland but is transported to the mouth to breakdown starch into maltose. Enzyme Action: ‘LOCK AND KEY’ HYPOTHESIS Each enzyme molecule has a very precise three dimensional shape. On part of its surface is a precisely shaped ‘dent’ known as the active site where the substrate molecule(s) will fit in exactly. The active site has to have a complementary (mirror image) to the shape of the substrate molecule(s). The enzyme is the ‘lock’ and the substrate is the ‘key’ as they work in the same manner as a lock and key i.e. key has to go towards the lock, which has a shape specific for that particular lock (=specific shape of active site). In catabolic (breaking down) reactions, large molecules are broken down into simpler ones: 1. When the substrate molecule is in position in the active site, the enzyme slightly stresses or bends the substrate, splitting it into 2 product molecules. 2. The product molecules move out of the active site leaving the active site free to combine with new substrate molecules. 3. It if involves water to split the substrates into products, this is known as hydrolysis. 4. Examples of catabolic reactions are digestion of food and respiration. sufe/bio/mspsbs/2012 Page 2 of 5 In anabolic (building up) reactions, large, complex substances are formed from small simple molecules. Examples of anabolic reactions are cell division, photosynthesis, synthesis of proteins and fats and repair of cells. Factors affecting enzyme activity 1. Temperature All enzymes will have an optimum working temperature, usually close to that at which they normally function. It is approximately 40 – 450C for most enzymes. In humans, the normal body temperature 370C is the optimum temperature where enzymes are most effective and rate of enzyme reaction is maximum. A rise in temperature increases the rate of activity of an enzyme. In many cases, a 10OC rise, will double the rate of reaction in a cell. The figure shown is a graph of enzyme activity against temperature. 1. Enzymes are inactive at very low temperature as the enzymes do not have enough kinetic energy and become inactivated. 2. As temperature rises, so does the rate of reaction. 3. The increase in enzyme activity is due to the increase in the kinetic energy causing the enzyme and substrate molecules to move faster. 4. This results in more successful collisions between enzyme and substrate molecules i.e. more substrates can enter the active site of an enzyme within a unit time to form products. 5. Also, the product molecules leave the active site of the enzyme more quickly. 6. At a certain temperature, a peak rate of activity is reached and this is known as the optimum temperature (temperature at which the enzyme works best). sufe/bio/mspsbs/2012 Page 3 of 5 7. When the temperature exceeds the optimum temperature (more than 500C), the rate of activity decreases rapidly to 0. a. This is because at these temperatures, the very high heat energy causes atoms of the enzyme molecule to change position relative to one another breaking the bonds holding them together. b. As a result, there is a change in the shape of the active site of the enzyme. c. The enzymes can no longer do their job since substrate molecules can no longer fit into their active site. d. At this stage, the enzymes are described as denatured (NOT killed as they were never alive). e. This damage is usually irreversible (permanent). 2. pH Most enzyme activities are affected by the pH of the solutions in which enzymes act. The optimum pH is the pH at which the rate of the enzyme activity is fastest. If the pH level falls on either side of the optimum, the rate of enzyme activity gradually decreases also due to denaturation of the enzyme molecules (unsuitable pH changes the shape of the enzyme molecule, hence changes the shape of the active site). Damage caused by pH is reversible, provided that the normal pH is restored as soon as possible. For most enzymes in a mammal’s body, the optimum pH is 7 or slightly above (slightly alkaline). Each enzyme molecule has a very precise three dimensional shape. On part of its surface is a precisely shaped ‘dent’ known as the active site where the substrate molecule(s) will fit in exactly. The active site has to have a complementary (mirror image) to the shape of the substrate molecule(s). sufe/bio/mspsbs/2012 Page 4 of 5 Uses of enzymes in daily life and industry Enzymes are produced abundantly from bacteria, fungi, plants and animals. They are used extensively at home as well as in industries. APPLICATION Meat tenderizer ENZYMES Protease USES Tenderize meat Protease Laundry detergent Lipase Breakdown protein molecules such as blood and dirt. Breakdown and removes oil and fatty stains. - Food processing: Production of chocolates, syrups, fruit juices and bread Meat and fish products in the food canning industry Amylase Protease Lactase Dairy products Removes starch in cocoa during chocolate production. - Converts starch to sugar in the making of syrup and fruit juices. - Changes starch flour to sugar during the making of bread. - Tenderize meat. - Removes the skin of fish before canning it. Changes lactose to glucose and galactose in the making of ice cream. Lipase Removes the fat part of milk to make cheese Leather industry Trypsin Removes hairs from animal skin Medical products Trypsin - Textile industry Amylase Removes starch which is used as stiffeners in fabrics. Treats swelling of wounds. Pre-digests baby food. Removes blood clots. A summary of enzymes involved in digestion in the human alimentary canal: REGION OF DIGESTION SECRETION SOURCE ENZYME ACTION MOUTH Saliva Salivary Glands Salivary Amylase Starch maltose STOMACH Gastric juice Gastric glands Pepsin Proteins polypeptides Rennin Soluble caseinogens insoluble casein Amylase Starch maltose Trypsin Proteins polypeptides Lipase Fats fatty acids and glycerol Sucrase Sucrose glucose + fructose Maltase Maltose glucose Erepsin Polypeptides amino acids Lactase Lactose glucose + galactose Lipase Fats fatty acids and glycerol Pancreatic juice Pancreas SMALL INTESTINES Intestinal juice sufe/bio/mspsbs/2012 Intestinal glands Page 5 of 5
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