How can we determine the effect of pH on amylase activity?

  • Thread starter PRINCESS NOOR
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In summary, the conversation discussed investigating the effect of pH on the activity of amylase in the human alimentary canal. The pH of the canal varies between pH 2 and pH 8.5 and iodine can be used to test for the presence of starch. The conversation also mentioned using spectroscopic experiments to set up the experiment, and provided some resources for conducting the assay.
  • #1
PRINCESS NOOR
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In the human alimentary canal, starch is broken down by amylase. The pH of the alimentary canal varies batween pH 2 and pH 8.5.

My question is how we can ivestigate the effect of pH on the activity of the amylase.

thank you.
 
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  • #2
With iodine you can test for the presence of starch by looking at the color, you could do spectroscopic experiments.
 
  • #3
experiment

How can we set this experiment?
 
  • #4
Thank you,

but how can we set this experiment?
 
  • #5
A PubMed search for "method for amylase activity spectrophotometry" brought up numerous hits, here are some. Pick one that looks good and get it or do the search and choose your own. If you have questions about the assay, I'm sure the board can help.

1: Dahot MU, Saboury AA, Moosavi-Movahedi AA.
Inhibition of beta-amylase activity by calcium, magnesium and zinc ions
determined by spectrophotometry and isothermal titration calorimetry.
J Enzyme Inhib Med Chem. 2004 Apr;19(2):157-60.
PMID: 15449730 [PubMed - in process]

2: Yasmineh WG, Chung MY, Caspers JI.
Determination of serum catalase activity on a centrifugal analyzer by an
NADP/NADPH coupled enzyme reaction system.
Clin Biochem. 1992 Feb;25(1):21-7.
PMID: 1551237 [PubMed - indexed for MEDLINE]

3: Soor SK, Hincke MT.
Microplate reader-based kinetic determination of alpha-amylase activity:
application to quantitation of secretion from rat parotid acini.
Anal Biochem. 1990 Jul;188(1):187-91.
PMID: 2171376 [PubMed - indexed for MEDLINE]

4: Sion JP, Laureys M, Gerlo E, Gorus F.
Detection of macroenzymes in serum by high-performance gel permeation
chromatography.
J Chromatogr. 1989 Nov 10;496(1):91-100.
PMID: 2592519 [PubMed - indexed for MEDLINE]

5: Satomura S, Sakata Y, Omichi K, Ikenaka T.
Alpha-amylase assay with use of a benzyl derivative of p-nitrophenyl
alpha-maltopentaoside, BG5P.
Clin Chim Acta. 1988 Jun 15;174(3):315-23.
PMID: 3134147 [PubMed - indexed for MEDLINE]

6: Rauscher E, Neumann U, Schaich E, von Bulow S, Wahlefeld AW.
Optimized conditions for determining activity concentration of alpha-amylase in
serum, with 1,4-alpha-D-4-nitrophenylmaltoheptaoside as substrate.
Clin Chem. 1985 Jan;31(1):14-9.
PMID: 3871178 [PubMed - indexed for MEDLINE]
 

What is the pH of the alimentary canal?

The pH of the alimentary canal can vary throughout different sections, but on average it ranges from 1.5 to 7.0. The stomach has the lowest pH of around 1.5-3.5 due to the presence of hydrochloric acid, while the small intestine has a more neutral pH of 7.0 due to the secretion of bicarbonate ions.

How does the pH of the alimentary canal affect digestion?

The pH of the alimentary canal plays a crucial role in digestion. Enzymes responsible for breaking down food have specific pH ranges in which they are most active. For example, pepsin in the stomach works best at a low pH while pancreatic enzymes in the small intestine work best at a neutral pH. Therefore, the varying pH levels throughout the alimentary canal support the efficient breakdown of different types of food.

What factors can affect the pH of the alimentary canal?

Several factors can influence the pH of the alimentary canal, including diet, stress, and medication. Consuming acidic or alkaline foods can alter the pH levels in the stomach, leading to digestive issues. Stress can also impact the secretion of stomach acid, which can affect the overall pH of the alimentary canal. Additionally, certain medications, such as antacids, can directly change the pH of the stomach.

How does the body maintain the pH balance in the alimentary canal?

The body has several mechanisms in place to maintain the pH balance in the alimentary canal. The stomach lining secretes mucus and bicarbonate ions to protect the stomach from the acidic environment. The pancreas also releases bicarbonate ions into the small intestine to neutralize the acidic chyme coming from the stomach. The liver and kidneys also play a role in regulating the body's overall pH balance.

What happens if the pH of the alimentary canal is too high or too low?

If the pH of the alimentary canal becomes too high or too low, it can disrupt the normal digestive processes. An excessively high pH can lead to the overgrowth of bacteria and impair the breakdown of food. On the other hand, a low pH can cause irritation and damage to the lining of the alimentary canal. In severe cases, an imbalance in pH can lead to conditions such as acid reflux, ulcers, or malabsorption of nutrients.

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