Module 6 Enzymes

Objectives To learn what enzymes are, why there are so many different ones in the body, why they are essential to life, and how they classified according to the types of reactions they are catalyze.

Introduction Enzymes are a type of protein: they act as catalysts for the many chemical reactions that take place in the body. Some enzymes are simple proteins and others are conjugated (contain nonprotein parts). In many cases these nonprotein parts of the enzyme are actually the active site in the catalysis process and often can be separated from the protein part. In these cases the nonprotein part is called a coenzyme. Each chemical reaction that takes place in the body is catalyzed by a specific enzyme. Approximately 1000 enzymes have been identified. Without these enzymes the chemical reactions would not take place at a fast enough rate to maintain life. Thus, the absence of even one of these enzymes in the body can have serious consequences. Since enzymes are proteins, high temperatures will cause them to become inactive. Thus, a temperature much above normal body temperature of 37o C ( 98.6oF) can be harmful to the enzymes in the body. Also, the activity of enzymes is dependent

on pH. Each enzyme has a particular pH range for each particular enzyme is quite small.

Discussion These are organic chemical substances produced by the activity of the living organisms such as plants, animals and microorganisms which modify the speed of a reaction without being used up or appearing as one of the end products.

Physical Nature o in solution, they are colloidal and non-dialyzable o soluble in water, glycerol and dilute alcohol o they are precipitated by protein precipitants like conc. Alcohol, ammonium sulfate and trichloroacetic acid.

Biological Importance o most chemical reactions characteristic of living organisms are accelerated by enzymes o it is estimated that a given cell could contain a thousand different enzymes, practically for every organic compound which occur in nature there exist also an enzyme capable of reacting with it. o These are responsible for the different reactions in living matter- systhesis, oxidation, hydrolysis, tautomeric changes- necessary during total metabolic reactions.

Naming ( Nomenclature ) of Enzymes: A. Upon the substance acted upon. The suffix ASE is added to the name of the substance acted upon. Examples: 1. Amylase acts on maltose ( amylum ) 2. Maltase acts on maltose 3. Lipase acts on lipids ( fats and lipids)

B. Upon the reaction enhanced. Also the suffix ASE is added to type of reaction catalyzed. Example: 1. Oxidase for oxidation 2. Decarboxylase for removal of CO2 3. Dehydrase/hydrase for reversible removal/ addition of water

C. The first enzymes discovered and studied were given common names based on the place of action/source. Example: 1. Ptyalin ( in saliva) 2. Trypsin ( in pancreatic juice) 3. Pepsin ( in gastric juice)

Classification of Enzymes: A. Hydrolases: enzymes that bring about hydrolysis reactions where a molecule is split into two or more smaller molecules by the addition of water. Common examples are: Proteases splits protein molecules. Examples: HIV protease and caspase. Nucleases splits nucleic acids (DNA and RNA). Based on the substrate type, they are divided into RNase and DNase. RNase catalyzes the hydrolysis of RNA and DNase acts on DNA. They may also be divided into exonuclease and

endonuclease. The exonuclease progressively splits off single nucleotides from one end of DNA or RNA. The endonuclease splits DNA or RNA at internal sites. Phosphatase catalyzes dephosphorylation (removal of phosphate groups). Example: calcineurin. The immunosuppressive drugs FK506 and Cyclosporin A are the inhibitors of calcineurin.

1. Esterases - is a hydrolase enzyme that splits esters into an acid and an alcohol in a chemical reaction with water called hydrolysis. 2. Glycosidases - catalyze the hydrolysis of the glycosidic linkage to generate two smaller sugars. They are extremely common enzymes with roles in nature including degradation of biomass such as cellulose and hemicellulose. 3. Amidases - is an enzyme that catalyzes the hydrolysis of an amide. 4. Peptidases - An enzyme that catalyzes the hydrolysis of peptides into amino acids. 5. Deaminases - an enzyme that hydrolyzes amino compounds (as amino acids) with removal of the amino group

B. Desmolases:

enzymes that add or remove specific group(s) from substances. 1. Dehydrases, hydrases 2. Desulfhydrases 3. Decarboxylases (non- oxidative) 4. Isomerases - is an enzyme that catalyzes the structural rearrangement of isomers. Isomerases thus catalyze

reactions of the form

where B is an isomer of A. 5. Aldolases - is an enzyme that helps convert glucose into energy. It is found throughout the body but is primarily found in high levels in muscle tissue. It is elevated in the bloodstream when a patient has muscle or liver damage or disease. 6. Acetylases addition of acetyl grouos 7. deaminases

C. Transferrases:

enzymes that transfer specific groups from one compound to another.

1. Transglycolases - an enzyme that catalyzes the transfer of glycosyl groups from one compound to another. 2. Transamidases - or an aminotransferase is an enzyme that catalyzes a type of reaction between an amino acid and an -keto acid. 3. Transpeptidases - an enzyme that catalyzes the transfer of an amino acid residue or a peptide residue from one amino compound to another 4. Trnasphosphorylases 5. Transmethylases - any of several enzymes that catalyze the

transfer of methyl groups from one compound to another. 6. Transformylases 7. Transforminases 8. Transhydroxymethylases 9. Transaminases 10. Transaldolases 11. Transacylases

D. Oxido-reductases:

enzymes that bring about oxidation and reduction reactions.

1. Oxidases - is any enzyme that catalyzes an oxidation-reduction reaction involving molecular oxygen (O2) as the electron acceptor. In these reactions, oxygen is reduced to water (H2O) or hydrogen peroxide (H2O2). 2. Anaerobic dehydrogenases 3. Aerobic dehydrogenases 4. Flavoprotein transhydrogenases 5. Transelectronases 6. Hydroxyperoxidases a. Peroxidase - enzymes degrade hydrogen peroxide, a naturally occurring byproduct of oxygen metabolism in the body. As a result, this substance is converted into water and oxygen.

b. Catalases - is a common enzyme found in nearly all living organisms that are exposed to oxygen, where it functions to catalyze the decomposition of hydrogen peroxide to water and oxygen.

Terminologies in enzymes chemistry 1. Substrate the substance acted upon 2. Zymogen/ proenzyme inactive form of enzyme 3. Activator - make enzymes work harder and faster 4. Co-enzyme 5. Apo-enzyme - a protein that combines with a coenzyme to form an active enzyme 6. Holo-enzyme 7. Anti-enzyme

Mechanism of Enzyme Action: 1. Simple Enzyme Action/Enzyme-Substrate Combination 2. Complex Enzymes Action/Enzymes- Co-enzyme System

Characteristics of Enzyme Action A. High Degree of Specificity 1. Absolute

2. Relative 3. Stereochemical

B. Reversibility of Enzyme Action C. Enzyme activity is maximal at an optimum temperature D. Enzyme have optimum pH for maximal activity

Factors that effect the rate of enzyme action: A. Physical Factors 1. Concentration of the substrate 2. Concentration of the enzyme 3. Effect of pH 4. Effect of temperature 5. Concentration of co-factors 6. Products of the reaction 7. Time 8. Radiation

B. Chemical Factor 1. Competitive Inhibition 2. Non-competitive Inhibition a. Oxidation-reduction effect

b. Formation of substrate resulting from the action of the inhibitors on the co-factors of the enzyme c. Heavy metals

Uses of Enzyme A. In Medicine Enzymes have become important tools in medicine. Today, enzymes are used to diagnose and to treat diseases. For example, enzyme assays of the blood serum reveal a great deal of information regarding the health of an individual. Frequently, the physicians use the enzyme assays to confirm their diagnoses from a set of symptoms that they have observed. If a person has chest pains and is suspected of having a myocardial infarction ( heart attack), an enzyme assay is used to verify if it is indeed a myocardial infarction and to determine the severity of the attack. When cells are damaged or die, an

abnormal amount of their enzymes leak out of the cells and enter the blood. Verification of myocardial infarction is accomplished by monitoring the blood concentrations of three enzymes namely: a. Creatine kinase, CK b. Serum-glutamate-oxaloacetate transaminase, SGOT c. Lactate dehydrogenase, LD

B. In Food Industry

Enzymes are added to foods to give them desirable properties not found in natural foods. Example: 1. Meat tenderizers normally contain papain, a protease derived from papaya plant. Papain hydrolyzes CHONs in tough cuts meat and makes them more tender. Sometimes, papain is

added to beers to hydrolyze CHONs that cause a haze in beer when it is chilled. 2. Invertase, a carbohydrate, is used to produce chocolate candies that have soft center filled with cherries and other fruits. Invertase and sucrose are added to the solid cherry before the chocolate coating is applied. Within a week to 10 days, the invertase hydrolyzes the sucrose and other carbohydrates in the cherry to more soluble (softer) and sweeter monosaccharides.

Enzymes in Food: Enzymes Corn Amylases Foods syrup in Function beer Increases the amount of

production, bakery goods, sugars and dextrins which cereals makes sweeter Remove glucose that the products

Glucose

Powdered

eggs,

cake interacts with albumin that

Oxidase

mixes

causes unpleasant

it

to

taste

Prevents chilled fruit drinks Pectinases Fruit and fruit drinks from becoming cloudy

Coagulates the milk protein Rennin Cheese production casein to produce cheese.

Enzyme-Deficiency-Related Diseases: It is an inherited condition in which a person lacks or has a small amounts of an important enzyme. Disease Albinism Missing Enzyme tyrosinase Physiological Problems Lack of pigmentation Ligament, cartilage, and Alkaptunuria Oxidase bone problems Mental retardation and Gauchers disease Beta-glucosidase early death Muscle glycogen is not converted to lactose, McArdlers disease phosphodylase causing muscle cramps during exercise

Mantal retardation and Niemans-Pick disease sphingomylinase early death Accumulation of Tay-Sachs disease hexosaminidase gangliosides in the brain, followed by early death Hydroxyphenyl-pyruvate tyrosinemia oxidase Liver and kidney defects

ASSIGNMENT 6 I. MULTIPLE CHOICE

1. Which of the following enzymes would digest a fat? a. Sucrose b. Fatase c. protease d. lipase

2. Enzymes are a special type of a. carbohydrates b. lipids c. proteins d. inorganic compounds

3. At high temperatures, the rate of enzyme action decreases because the increased heat a. changes the pH of the system b. alters the active site of the enzyme c. neutralizes the acids and bases in the system d. increases the concentration of the enzyme

4. Enzymes influence chemical reactions in living systems by a. providing the substrate required for the reaction to occur b. affecting the rate at which reactions occur c. absorbing water released when polymers are formed d. combining with excess hydrogen to form gaseous wastes 5. Which group of organic compounds includes the enzymes? a. Proteins b. Starches c. Carbohydrates d. lipids

6. The "lock and key hypothesis" attempts to explain the mechanism of a. vacuole formation b. pinocytosis c. sharing of electrons d. enzyme specificity

7. Any substance that is acted upon by an enzyme is called a. Coenzyme b. Substrate c. Vitamin d. polypeptide

8. An enzyme that hydrolyzes protein will not act upon starch. This fact is an indication that enzymes are a. Hydrolytic b. specific c. catalytic d. synthetic

9. At about 0 C., most enzymes are a. Inactive b. Active c. Destroyed d. replicated 10. Vitamins are essential to the survival of organisms because vitamins usually function as a. Substrates b. nucleic acids

c. coenzymes d. nucleotides 11. Which chemical is classified as an enzyme? a. Galactose b. lipid c. protease d. manganese dioxide

12. Salivary amylase is an enzyme in humans that breaks down starch. The optimum pH for this reaction is 6.7. The rate of this reaction would not be affected by a. maintaining the pH of the reaction at 6.7 b. substrate concentration c. enzyme concentration d. decreasing the temperature of the reaction by 5 degrees C

13. A certain enzyme will hydrolyze egg white but not starch. Which statement best explains this observation? a. Starch molecules are too large to be hydrolyzed. b. Enzyme molecules are specific in their actions. c. Egg white acts as a coenzyme for hydrolysis d. Starch is composed of amino acids.

14. Which environmental condition would most likely have the LEAST effect on the rate of enzyme controlled hydrolytic reactions in humans? (1.) (2.) (3.) (4.) a. the pH of the solution b. the temperature of the solution c. the amount of enzyme present d. the amount of light present

15. Which statement best expresses the information represented in the graph shown? a. The action of enzymes varies with pH. b. A pH of 7 provides the optimum environment for digestive enzymes c. Gastric juice is active at a pH extending from 0 to 12. d. Acids have a pH greater than 7.

16. Lipase, maltase, and protease are members of a group of catalysts known as a. Hormones b. Carbohydrates c. Lipids d. enzymes

17. The optimum temperature for the action of this enzyme is approximately a. 15 C b. 22 C

c. 37 C d. 50 C

18. The fact that amylase in the human small intestine works best at normal body temperature (37 degree C) suggests that a. amylase is denatured at temperatures below 37 degrees C b. amylase can function only in the small intestine c. the lock-and-key model of enzyme action does not apply to amylase d. the optimum temperature for amylase is 37 degrees C

19. A catalyst is a substance which a. increases the rate of a chemical reaction but is itself unchanged by the reaction b. is toxic to most cells c. is composed mostly of lipids d. does not usually participate in any chemical reactions

20. Coenzymes are best defined as a. organic molecules which can serve as substitutes for enzymes b. inorganic molecules which can serve as substitutes for enzymes c. organic molecules which help in enzyme-catalyzed reactions by bonding with electrons

d. inorganic molecules which help in enzyme-catalyzed reactions by bonding with electrons

II.

Answer the ff: 1. State how the following will affect the rate of an enzyme reaction. a. increasing temperature b. decreasing temperature c. pH d. salt concentration

2. Briefly describe a generalized enzyme-substrate reaction, state the function of an enzyme's active site, and describe how an enzyme is able to speed up chemical reactions.