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

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Discussion Overview

The discussion centers on investigating the effect of pH on the activity of amylase, an enzyme involved in starch breakdown in the human alimentary canal. Participants explore potential experimental setups and methods for measuring amylase activity across varying pH levels.

Discussion Character

  • Experimental/applied
  • Technical explanation
  • Homework-related

Main Points Raised

  • One participant notes that the pH in the human alimentary canal ranges from pH 2 to pH 8.5 and poses a question about how to investigate amylase activity under these conditions.
  • Another participant suggests using iodine to test for starch presence, indicating that color changes could be monitored, and mentions the possibility of conducting spectroscopic experiments.
  • Several posts inquire about how to set up the experiment, indicating a need for clarity on the experimental design.
  • A participant provides a list of PubMed articles related to methods for measuring amylase activity using spectrophotometry, suggesting that others could select a relevant method for their investigation.

Areas of Agreement / Disagreement

Participants express a general agreement on the need for an experimental approach to investigate the effect of pH on amylase activity, but there is no consensus on a specific experimental design or method to be used.

Contextual Notes

The discussion lacks detailed assumptions about the specific conditions under which the experiments would be conducted, such as temperature or enzyme concentration, which may affect the outcomes. There are also unresolved questions regarding the selection of appropriate methods from the provided literature.

Who May Find This Useful

Students and researchers interested in enzymology, particularly those studying the effects of pH on enzyme activity, may find this discussion relevant.

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

How can we set this experiment?
 
Thank you,

but how can we set this experiment?
 
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]
 

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