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BIOL 204 Lab For Week 12 Digestive Enzymes

BIOL 204 Lab For Week 12 Digestive Enzymes. Digestive Enzymes : are used in the lumen of the GI tract to break down complex molecules into absorbable subunits

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BIOL 204 Lab For Week 12 Digestive Enzymes

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  1. BIOL 204 Lab For Week 12 Digestive Enzymes

  2. Digestive Enzymes: • are used in the lumen of the GI tract to break down complex molecules into absorbable subunits • Enzymes are biological catalysts which increase the rate of a chemical reaction without themselves becoming part of the product:

  3. In this experiment we will observe the activity of three different enzymes: Amylase:digests starch into disaccharides Trypsin:digests proteins into peptides Lipase: digests fats into glycerol and fatty acids

  4. Variables tested …… • Incubation temperature: • How will enzyme activity compare when incubated at O o C and 37 o C? • Denaturization of Enzymes • How will enzyme activity be altered if the enzyme is denatured by boiling?

  5. Activity 1 Trypsin Activity Trypsin an enzyme, secreted by pancreatic acinar cells, that hydrolyzes proteins into small fragments (peptones, peptides and proteoses)”

  6. In this experiment, trypsin will act on an artificial substrate to yield a yellow colored product: Trypsin BAPNA + H2O N-benzoyl-L-arginine + p-nitroaniline (yellow) 1. Reaction rate will be compared at these different incubation temperatures: 370 C and 0o C What do you predict will happen? 2. The rate of the reaction will be determined using boiled trypsin. What do you predict will happen?

  7. Activity 2 Amylase Activity Amylaseis a carbohydrate digesting enzyme, produced by salivary glands and pancreatic acinar cells, that hydrolyzes starch into maltose (disaccharides): Amylase Starch + H2O maltose (disaccharides)

  8. Indicators: Starch: IKI (Lugol’s solution) - turns “blue/black” in the presence of starch Benedict’s Solution: turns green/orange/ red/brown in the presence of reducing sugar

  9. IKI Test For Starch in a Spot Plate Benedict’s Test for reducing sugar in heat test tubes

  10. As in activity one, the reaction will be observed under different conditions: Boiled Enzyme (Amylase) Incubation Temperature: 0oC and 37oC

  11. Activity 3 Lipase Activity Lipase is a hydrolytic enzyme, made primarily by pancreatic acinar cells, that hydrolyzes neutral fats into fatty acids and glycerol: Lipase Fat + H2O Glycerol + Fatty Acids

  12. Fat Digestion: Indicator: Blue litmus terns pink (red) in the presence of acid (one the products is fatty acids)

  13. Bile emulsifies fat….

  14. In this activity, you will examine the effect of: incubation temperature (0oC vs. 37oC) and addition of bile salts on the reaction

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