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Lab 5: Amylase Enzyme Activity

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Melinda Chisholm

on 31 March 2015

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Transcript of Lab 5: Amylase Enzyme Activity

Lab 5: Amylase Enzyme Activity
and Action of Inhibitors

Interesting Facts and Extra Research!
Amylase breaks down starch, a storage sugar molecule for glucose found in plants. It is also capable of performing the same function on glycogen, the starch equivalent found in humans, and breaks it down into simple sugars (so not necessarily glucose).

Amylase is used therapeutically in its regulation of histamine, which is produced by cells as a response to recognized invaders in the body. Histamine is produced in allergic reactions and is what causes hives, itchy and watery eyes, and sneezing and runny noses. This means that amylase is effective in breaking down the histamine produced by the body in response to allergens.

Amylase is predominantly found in the human body as either pancreatic amylase or salivary amylase, although it is found in many tissues. Salivary amylase breaks down starch into maltose and dextrin, and is deactivated in the stomach by gastric acid. Pancreatic amylase randomly cleaves the glycosidic linkages of amylose, yielding dextrin, maltotriose, and maltose.
Works Cited
Biology 107 Laboratory Manual. 2014-2015. Lab 5, Amylase Enzyme Activity and Action of Inhibitors, p5-5 - 5-9, University of Alberta, Edmonton, Alberta, Canada.

Howell, Dr. Edward. "Amylase - They're Called Amylolytic Enzymes." Enzyme-Facts. 2009. Web.

Lab 5 Summary
In the experiment for lab 5, we measured enzymatic activity of amylase on starch. The enzyme and substrate were mixed, then separated into tubes and examined at three-minute intervals for a total of 12 minutes. Iodine was added to each of the solutions in order to act as an indicator to the presence of starch, and absorbance of the solution was measured at each three-minute interval using a spectrophotometer. Greater absorbance values correspond to a higher concentration of starch present within the solution, and lower absorbance values correspond to a lower concentration of starch. Through performing this experiment, we found that in the presence of amylase, starch concentrations decreased over time, inferring that amylase breaks down starch.

Our Question:
A) What does the enzyme amylase do to amylose, a component found within starch molecules that allows for it to pass through the cell membrane?

B) If there was a mutation within the genome that codes for amylase, what would happen if it were to be a silent mutation, missense, or nonsense mutation?

Interesting Facts and Extra Research (Continued)
There is genetic variation present within the salivary amylase produced by humans. It appears that the gene that codes for salivary amylase has undergone duplication during evolution due to high-starch diets. Perry, et al. published findings in an article “Diet and evolution of human amylase gene copy number variation”, in Nature Genetics (2007). They found that an individual living in Japan had 14 copies of the amylase gene, while a Biaka individual had 6 copies. Biaka are rainforest hunter-gatherers with a low starch diet.
Interesting Facts and Extra Research (Continued... Again!)
Amylase testing is the primary method used to detect and monitor pancreatitis, an inflammation of the pancreas that can ultimately result in bleeding into the gland, serious tissue damage, infection, cyst formation, and pancreatic failure. Increased plasma levels of amylase in humans are found in: the mumps (an inflammation of the salivary glands); salivary trauma such as anaesthetic intubation (the insertion of a tube into the trachea in order to maintain respiration or to administer medication); pancreatitis due to damage to cells producing the enzyme; and renal failure due to reduced excretion. Pancreatitis usually produces total amylase readings ten times greater than the upper limit of normal. Duodenal disease or renal failure produces amylase five to ten times greater than the upper limit of normal, and lower elevations are common in salivary gland disease.
A) Amylase reduces starch into smaller glucose molecules so that they may pass through the cell membrane or be transported more easily into the cell. The permeability of the cell membrane also depends on the properties of the molecules, including whether they are hydrophilic or hydrophobic in nature.

B) A silent mutation would have no effect on the enzyme. The enzyme would be created properly and still capable of functioning as required. A missense mutation would cause some problems because for a short period of time, the enzyme would be incorrectly coded for. A nonsense mutation would result in the genome encoding different proteins, and amylase would not be produced.
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