Enzymes: Structure, Function and Classification

ENZY MES Topics for Discussion ✗ What is enzyme? ✗ Uses, functions and effects ✗ Foods with High Content of Enzyme ✗ Pros and Cons ✗ Ways to Analyze its Presence in Food ✗ Value on Nutrition Facts ✗ Chemical Properties ✗ FDA Requirements and 2 Classifications  ENZYMES ✔ Protein...

ENZY MES Topics for Discussion ✗ What is enzyme? ✗ Uses, functions and effects ✗ Foods with High Content of Enzyme ✗ Pros and Cons ✗ Ways to Analyze its Presence in Food ✗ Value on Nutrition Facts ✗ Chemical Properties ✗ FDA Requirements and 2 Classifications  ENZYMES ✔ Protein molecules (made up of amino acids) ✔ Biological catalysts (increase the speed of chemical reaction) ✔ Specific ( each one catalyses one particular reaction)  ENZYMES ✔ Reusable (can be used again and again) ✔ Affected by temperature and pH ✔ Found in animals, plants and microorganisms STRUCTURE OF ENZYMES ❑ Enzymes are proteins ❑ They have a globular shape ❑ A complex 3d structure 5  CHARACTERIS TICS OF ENZYMES ✔ Enzymes are unique in nature and show varying degree of specificities, while they are highly specific for a particular substrate. ✔ Enzymes are extremely efficient and possess great catalytic power ✔ Enzymes itself remain unchanged during the reaction  CHARACTERIS TICS OF ENZYMES ✔ Enzymes can be allosterically regulated by a variety of means, but the activity of the catalysts cannot be controlled ✔ The effectiveness of an enzyme catalyst could achieve maximum at its optimum temperature ✔ Biochemical catalysis of enzyme also depends on the pH of the solution CHARACTERIS TICS OF ENZYMES ✔ The catalytic activity of enzymes can be inhibited by competitive inhibitors, noncompetitive inhibitors or irreversible inhibitors. ✔ Increase in the concentration of the reactants could increase the reaction rate until the enzyme become saturated with the substrate, while increase in the amount of enzyme will continuously enhance the rate of the reaction. CHARACTERIS TICS OF ENZYMES ✔ Enzymes could function reversely, signifying that the enzyme does not determine the direction of reaction. It only functions to accelerate reaction rate until equilibrium is achieved. 2 MAIN TYPES OF ENZYMES ❑ CATABOLIC ENZYMES which break down large molecules into smaller molecules ❑ ANABOLIC ENZYMES which build up small molecules to form larger molecules 10 CLASSIFICATION OF ENZYMES ✗ Food Enzymes– occur in raw food and, when present in diet, begin the process of digestion. ✗ Digestive Enzymes – produced by the body to break food into particles small enough to be carried across the gut wall. ✗ Metabolic Enzymes - produced by the body to perform various complex biochemical processes. 11 TYPES OF DIGESTIVE ENZYME ✗ Amylase - breaks down starches and carbohydrates into sugars. ✗ Protease - breaks down proteins into amino acids. ✗ Lipase - breaks down lipids, which are fats and oils, into glycerol and fatty acids. 12 SIX FUNCTIONAL CLASSES OF ENZYME TYPES BIOCHEMICAL PROPERTY catalyzes the oxidation reaction where Oxidoreductases the electrons tend to travel from one form of a molecule to the other. help in the transportation of the Transferases functional group among acceptors and donor molecules. 13 SIX FUNCTIONAL CLASSES OF ENZYME TYPES BIOCHEMICAL PROPERTY catalyze the hydrolysis reaction by Hydrolases adding water to cleave the bond and hydrolyze it. adds water, carbon dioxide or ammonia across double bonds or Lyases eliminate these to create double bonds. 14 SIX FUNCTIONAL CLASSES OF ENZYME TYPES BIOCHEMICAL PROPERTY catalyze the structural shifts present in Isomerases a molecule, thus causing the change in the shape of the molecule. charge the catalysis of a ligation Ligases process. 15 MICROBIAL ENZYME Microbial enzymes are known to be superior enzymes obtained from different microorganisms, particularly for applications in industries on commercial scales. 16 EXAMPLES OF SPECIFIC ENZYMES ✗ Lipases – a group of enzymes that help digest fats in the gut. ✗ Amylase – helps change starches into sugars. Amylase is found in saliva. ✗ Maltase – also found in saliva; breaks the sugar maltose into glucose. ✗ Trypsin – found in the small intestine, breaks proteins down into amino acids. 17 EXAMPLES OF SPECIFIC ENZYMES ✗ Lactase – also found in the small intestine, breaks lactose, the sugar in milk, into glucose and galactose. ✗ Acetylcholinesterase – breaks down the neurotransmitter acetylcholine in nerves and muscles. ✗ Helicase – unravels DNA. ✗ DNA polymerase – synthesize DNA from deoxyribonucleotides. 18 what do enzymes do? ❑ THE DIGESTIVE SYSTEM enzymes help the body break down larger complex molecules into smaller molecules, such as glucose, so that the body can use them as fuel ❑ DNA REPLICATION which build up small molecules to form larger molecules ❑ LIVER ENZYMES the liver breaks down toxins in the body. To do this, it uses a range of enzymes. 19  LACK OF ENZYMES Lack of a certain enzyme will definitely interfere in the working of another and this makes a chain altogether. Enzyme deficiencies, or the absence of these enzymes, are inherited defects that result in a number of life-changing or life- threatening conditions: LACK OF ENZYMES ✗ MPS: ✗ The mucopolysaccharidoses are a group of inherited diseases in which a defective or missing enzyme causes complex sugar molecules to accumulate in cells. As a result, progressive damage is done to the heart, bones, joints, respiratory system and central nervous system. 21 LACK OF ENZYMES ✗ LSD: ✗ Lysosomal storage disorders are a group of approximately fifty inherited disorders that occur when a missing enzyme results in the body’s inability to recycle cellular waste. The severity of the disorder depends on the type and amount of cellular debris that accumulates, but almost all disorders are progressive. Many of these children die in infancy or early childhood. 22 LACK OF ENZYMES ✗ NP: ✗ Nieman-Pick Disease refers to a group of inherited metabolic disorders known as lipid storage disorders. Patients diagnosed with these disorders lack a critical enzyme necessary to metabolize fatty substances in the body called lipids. As a result, harmful quantities of lipids accumulate in the spleen, liver, lungs, bone marrow and brain. 23 USES, FUNCTION, AND EFFECTS ON FOOD ❑ Recovery of new ingredients ❑ Food modification ❑ Improving sensory quality - flavor development ❑ Partialy replacing some of the chemical additive ❑ Increasing yields ❑ Beverage clarification ❑ Bakery aids ❑ Meat tenderization 24 USES, FUNCTION, AND EFFECTS ON FOOD MICROBIAL ENZYME APPLICATION Baking, brewing, starch liquefaction Bread quality improvement α-Amylase Rice cakes Clarification of fruit juice Beer production Bread quality improvement Glucoamylase High glucose and high fructose syrups Lactose intolerance reduction in Lactase (β-galactosidase) people Prebiotic food ingredients 25 USES, FUNCTION, AND EFFECTS ON FOOD MICROBIAL ENZYME APPLICATION Food preservation (with glucose oxidase) Catalase Removal of hydrogen peroxide from milk prior to cheese production Development of flavour, colour and Peroxidase nutritional quality of food Removal of bitter taste in fruit juice Debittering enzymes - naringinase Wine aroma enhancement 26 USES, FUNCTION, AND EFFECTS ON FOOD MICROBIAL ENZYME APPLICATION Brewing Meat tenderization Protease Coagulation of milk Bread quality improvement Cheese flavour development Lipase Cheddar cheese production Pectinase Clarification of fruit juice 27  FOODS WITH HIGH CONTENT OF ENZYMES ❑ PINEAPPL ❑ PAPAYA ❑ MANGOE ❑ HONEY E S ❑ AVOCADO ❑ BANANA ❑ KIWIFRUI ❑ GINGER T 28 IN HUMANS Advantages Disadavantages □ Building muscle □ Inflamed or injured liver cells □ Destroying toxins □ Breaking down food particles during digestion 29 At home Advantages Disadavantages □ These are more □ If the clothes are not efficient at removing fully rinsed, protease stains from clothes. enzymes may remain in the clothes, which □ Lower washing digests protein in the temperatures can be skin. used which saves energy. □ This can lead to irritation, allergies and dermatitis. 30 In industry Advantages Disadavantages □ Supplying heat is □ Enzymes are sensitive expensive. When to temperature and pH enzymes are used, changes. industrial processes can be cheaper. □ Temperature and pH need to be carefully □ Fully biodegradable. monitored and controlled. 31 Determination of enzymes in food Enzyme assay ✗ Is a test conducted by an individual to determine their protein of interest based on its unique functionality and properties on a sample. 33 Enzyme activity ✗ Is measured in units which indicates the rate of reaction catalyzed by that enzyme. 34 35  2 types of assay Continuous Assay Discontinuous Assay -An assay which gives – the reaction proceeds continuous reading activity for a designated time, - multiple measurements, – the reaction is stopped usually of absorbance (usually by inactivating change, are made during the enzyme with a weak the reaction, acid), – either at specific time – a measurement is intervals (usually every 30 made of the amount of or 60 seconds) reaction that has occurred 36  Enzyme assay methods Continuous Assay Spectrophotometric Fluorescence method method/fluorimetric ✗ It uses a ✗ Uses a Fluorometer spectrophotometer, a ✗ Fluorometric assays machine used to measure use a difference in the the amount of light a fluorescence of substance's absorbs, to substrate from product combine kinetic measurements and Beer's to measure the enzyme law by calculating the reaction. appearance of product or 37 disappearance of  Enzyme assay methods Continuous Assay Calorimetric method Chemilluminescent ✗ is the measurement of ✗ is the emission of light by a the heat released or chemical reaction. ✗ Some enzyme reactions absorbed by chemical produce light and this can be reactions. measured to detect product ✗ These assays are very formation. general, since many ✗ These types of assay can be reactions involve some extremely sensitive, since the change in heat and with light produced can be use of a micro-calorimeter, captured by photographic film not much enzyme or over days or weeks, 38 substrate is required.  Enzyme assay methods Discontinuous Assay Radiometric method Chromatographic method ✗ Radiometric assays measure the ✗ Chromatographic assays incorporation of radioactivity into measure product formation by substrates or its release from substrates. separating the reaction ✗ The radioactive isotopes most mixture into its components frequently used in these assays are by chromatography. 14C, 32P, 35S and 125I. ✗ This is usually done by high- ✗ They are frequently used in biochemistry and are often the only performance liquid way of measuring a specific reaction chromatography (HPLC) in crude extracts (the complex mixtures of enzymes produced when you lyse cells) 39 Nutrition facts on food products According to CRN-ETA’s “best practices guide” on enzyme dietary supplement products: “For compliant and meaningful labels, it is recommended that enzyme-containing dietary supplements be labeled with both metric measures and activity units either by individual enzyme or as proprietary blends of enzymes.” 40 ✗ However, according to US-FDA, Lactose, as a sugar in a dairy product, is a disaccharide that is captured on the Nutrition Facts ✗ label under the Total Sugars declaration, and lactose that was obtained in whey should be labeled under the Added Sugars and Total Sugars declaration. 41 ✗ In enzyme preparation, there should contain an information about the presence of proteins that were derived from major food allergens under the provisions of Food Allergen Labelling and Consumer Protection Act of 2004 (FALCPA) 42 CLASSIFICATION OF ENZYMES ✗ Oxidoreductases - Catalyze oxidation-reduction reactions ✗ Transferases - Catalyze group transfer reactions ✗ Hydrolases - Catalyze hydrolysis reactions where water is the acceptor of the transferred group ✗ Lyases - Catalyze lysis of a substrate, generating a double bond in a nonhydrolytic, nonoxidative elimination ✗ Isomerases - Catalyze isomerization reactions 43 CLASSIFICATIONS OF ENZYMES 44 45 Allowable dosage 46  Sources https://www.google.com/url?sa=t&rct=j&q=&esrc=s&source=web&cd=&cad=rja&uact=8&ved=2ahUKEwijp 8Lc17fwAhUSfd4KHXtGD_IQFjAAegQIAxAD&url=https%3A%2F%2Fwww.fda.gov%2Fmedia%2F79379 %2Fdownload&usg=AOvVaw18-0Jjcb9eTDcTEVfJATg_ https://youtu.be/rgpN2OXg7mI https://www.google.com/url?sa=t&rct=j&q=&esrc=s&source=web&cd=&cad=rja&uact=8&ved=2ahUKEwiD j6_r17fwAhXQ62EKHdpYCW8QFjAAegQIBBAD&url=https%3A%2F%2Fwww.slideshare.net%2Fiamrahul sethi%2Fenzymes-assay-ppt-best&usg=AOvVaw2xZiHNgNjZ39v0mGVvIkjI https://meridian.allenpress.com/jfp/article-pdf/46/5/453/1650621/0362-028x-46_5_453.pdf https://www.google.com/url?sa=t&rct=j&q=&esrc=s&source=web&cd=&cad=rja&uact=8&ved=2ahUKEwiy pfC137fwAhUSfd4KHXtGD_IQFjAAegQIBhAD&url=https%3A%2F%2Fwww.fda.gov%2Fmedia%2F11740 2%2Fdownload&usg=AOvVaw1DC3pCKGVtMnpKcrqwgTcN 47  THANK YOU FOR LISTENING