Enzymes Notes

Enzymes

  • Proteins with catalytic properties.
  • Examples:
    • Lipase (lipids)
    • Protease (proteins)
    • Amylase (starch)

Objectives

  • Explain the role and importance of enzymes, including:
    • Catalysis
    • Properties of enzymes
  • Investigate the effect of temperature and pH on the activity of enzymes like catalase or amylase.

Experiment Time: Catalase and Hydrogen Peroxide

  • Materials:
    • Small beaker containing a slice of liver
    • Big beaker containing hydrogen peroxide
  • Procedure:
    • Carefully pour the hydrogen peroxide over the liver.
    • Observe what happens.
    • Note observations in notebooks.
  • Fun fact: Liver contains catalase, which breaks down hydrogen peroxide into oxygen and water.

What are Enzymes?

  • Biological catalysts.
  • Catalysts speed up chemical reactions.
  • Without enzymes, reactions in living organisms would be too slow to sustain life.
  • Enzymes are biological molecules that accelerate chemical reactions.

Importance of Enzymes

  • Without enzymes, reactions in cells would be too slow to occur, leading to the organism's death.
  • Enzymes control many reactions in the cell, allowing them to proceed in an orderly fashion.

Enzyme Structure

  • Enzymes are proteins.
  • They have a globular shape.
  • They possess a complex 3-D structure.
  • Enzymes have an active site.

Substrate and Active Site

  • Substrate: The substance/chemical (reactant) that the enzyme acts/works on.
    • Example: In an enzyme-catalyzed reaction to make glycogen, glucose would be the substrate.
  • Active site: The specific region on the enzyme to which the substrate binds to undergo the chemical reaction.
  • Enzymes catalyze the breakdown of a substrate into product(s).
  • Product: The substance that is made by the reaction.
    • Example: Breakdown of starch by amylase, the product is maltose.
  • A \rightarrow B + C (Substrate to Products)

The Substrate

  • Substrates are reactants activated by enzymes.
  • Enzymes are specific to their substrates.
  • Specificity is determined by the active site.
  • The enzyme name often corresponds to the substrate it breaks down.
    • Lipases catalyze the breakdown of lipids.
    • Carbohydrases catalyze the breakdown of carbohydrates.

Enzyme Mode of Action: The Lock and Key Hypothesis

  • Enzyme may be used again.
  • Enzyme-substrate complex (E+S -> E+P)
  • The shape and chemical environment inside the active site allow a chemical reaction to proceed more easily.
  • Enzyme + Substrate -> Enzyme-Substrate Complex -> Enzyme + Products

Lock and Key Hypothesis Details

  • The fit between the substrate and the active site of the enzyme is exact.
  • Like a key fits into a lock very precisely.
  • Enzyme-substrate complex is formed.
  • Products have a different shape from the substrate.
  • Products are released from the active site, leaving it free for another substrate molecule.

Lock and Key Diagram

  • Diagram showing the enzyme, substrate, active site, enzyme/substrate complex, and products.

Explanation of Enzyme Specificity and Denaturation

  • Explains enzyme specificity.
  • Explains the loss of activity when enzymes denature.
  • Enzyme denaturation: The process by which an enzyme’s structure is changed, and the active site loses its shape, thereby disabling the enzyme from functioning.

Properties of Enzymes

  • Made of protein, from amino acids produced by digestion or the chemical breakdown of other proteins.
  • Biological catalysts, not changed by the reactions they speed up, and can be used repeatedly.
  • Specific: Only one particular enzyme will work with one particular substrate.

Properties of Enzymes (Continued)

  • Temperature sensitive: Enzymes are denatured or inactivated by excess heat because they are protein. They work best at an optimum temperature.
  • Sensitive to pH (degree of acidity and alkalinity of a medium). They work best at an optimum pH.
  • Can be inhibited by poisons such as arsenic and cyanide.
  • Require moisture to operate, hence found inside our cells.

Factors Affecting Enzymes

  • Enzymes work best under certain conditions and are affected by:
    • Enzyme concentration
    • Substrate concentration
    • pH
    • Temperature
    • Inhibitors
  • When measuring one factor, keep all other factors constant.

The Effect of Temperature

  • Heating increases the kinetic energy of molecules.
  • Increased kinetic energy of the enzyme increases molecular motion.
  • This raises the chances of the enzyme colliding with a substrate.

The Effect of Temperature (Continued)

  • All enzymes have an optimum temperature that promotes maximum activity.
  • If the enzyme’s temperature is increased above its maximum, the enzyme becomes denatured, and enzymatic activity is lowered, potentially stopping.

The Effect of Temperature (Continued)

  • If the temperature becomes too low or cold, enzymes become inactivated but NOT denatured.
  • They can become activated again once an appropriate temperature is reached.

Temperature Scale and Enzyme Activity

  • Graph showing enzyme activity versus temperature.
  • Optimum temperature marked.
  • Denaturation point indicated.

The Effect of Temperature: Specific Examples

  • For most enzymes, the optimum temperature is about 30°C.
  • Mammalian enzymes tend to have a temperature range between 37-40°C.
  • Cold-water fish enzymes denature at 30°C.
  • Some bacteria have enzymes that withstand very high temperatures between 70-100°C.
  • Most enzymes are fully denatured at 70°C.

The Effect of pH

  • Different enzymes work at different pH conditions.
  • Under constant temperature and other conditions, most enzymes function efficiently over a narrow pH range.
  • If the pH is increased or decreased below an enzyme’s pH range, the enzyme activity will decrease.
  • When pH increases, this means alkalinity increases, and so does the concentration of hydrogen ions.

The Effect of pH (Continued)

  • Extreme pH levels will produce denaturation.
  • The structure of the enzyme is changed as the active site is distorted, and substrate molecules will no longer fit.
  • At extreme pH values, changes in the charges on substrate molecules will also occur.
  • This change will affect the binding of the substrate with the active site.

pH Scale

  • Diagram of the pH scale, ranging from strong acid to strong alkali.
  • pH values from 1 to 14.

The Effect of pH on Enzyme Activity Graph

  • Graph showing rate of reaction versus pH.
  • Optimal pH for pepsin (stomach enzyme) and trypsin (intestinal enzyme) are marked.

pH of the Alimentary Canal

  • Mouth: Slightly alkaline to neutral.
  • Stomach: Very acidic; pH 2 (dilute hydrochloric acid).
  • Duodenum: Very alkaline (bile).
  • Ileum: Alkaline.

Enzyme Inhibition

  • An inhibitor is a small molecule or chemical which reduces or stops the rate of the activity of an enzyme.
  • Inhibitors help to regulate enzyme activity.
  • Many drugs and poisons act as enzyme inhibitors.
  • Example: Cyanide inhibits an enzyme involved in respiration.

Enzyme Concentration

  • If other conditions are kept constant and substrate concentration is maintained at a high level, the rate of reaction is proportional to the enzyme concentration.

Substrate Concentration

  • The rate of an enzyme reaction increases with increasing substrate concentration for a given enzyme.
  • However, there comes a point when no increase in substrate concentration will cause a significant increase in enzyme reaction rate.
  • Faster reaction but it reaches a saturation point when all the enzyme molecules are occupied.

Substrate Concentration Explanation

  • At high substrate concentrations, the active sites of the enzyme molecules are occupied with substrate.
  • Therefore, a substrate would have to wait for a reaction to complete and the products to leave the active site before it can enter into the active site of the enzyme.

Catalase

  • An enzyme found in animal and plant cells.
  • Detoxification is one of the liver's functions.
  • Catalase is a natural enzyme found primarily in the liver of animals.
  • It splits hydrogen peroxide, which is toxic to cells, into harmless oxygen and water.
  • Needed to speed up the breakdown of hydrogen peroxide.
  • Breaks it down to oxygen and water.
  • Word equation: Hydrogen Peroxide -> Oxygen + Water

Amylase

  • Found in saliva and in the pancreas.
  • Break down enzyme.
  • Breaks starch down to maltose.
  • Word equation: Starch -> Maltose

More About Specificity

  • Amylase will only breakdown starch.
  • Catalase will only breakdown hydrogen peroxide.
  • Amylase will NOT breakdown hydrogen peroxide.
  • Catalase will NOT breakdown starch.
  • Are specific for what they will catalyze.
  • Are Reusable.
  • End in –ase (Sucrase, Lactase, Maltase).

Examples of Digestive Enzymes in the Alimentary Canal

  • Lipase: Breaks down fat to glycerol and fatty acids. Produced in the pancreas and small intestine.
  • Protease (e.g., pepsin, trypsin): Breaks down proteins to amino acids. Produced in the stomach, pancreas, and small intestine.
  • Carbohydrase (e.g., Maltase, Lactase, Sucrase): Breaks down carbohydrates to simple sugars. Produced in the small intestine and pancreas.
  • Salivary amylase: Breaks down polysaccharides to maltose. Produced in salivary glands.
  • Pancreatic amylase: Breaks down polysaccharides to maltose. Produced in the pancreas.
  • Catalase: Breaks down toxic hydrogen peroxide to water and oxygen. Produced in most tissues.

Uses/Applications of Enzymes

  • Enzymatic browning common in:
    • Fruits (Apples, pears, peaches, apricots, and bananas)
    • Vegetables (Potatoes, lettuce, brinjal)
    • Cereals (wheat flour, rice)
    • Sea foods (shrimps, spiny lobsters and crabs)

Uses/Applications of Enzymes: Buzzle.com

  • Substitutes for commercially produced meat tenderizer powder:
    • Meat mallet
    • Papaya paste
    • Pineapple juice

Uses/Applications of Enzymes: Laundry Detergent

  • Enzymes used in laundry detergent for stain removal.