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

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The discussion focuses on investigating the effect of pH on amylase activity in the human alimentary canal, where starch is broken down by amylase. The pH levels in this canal range from 2 to 8.5. To explore this, participants suggest using iodine to test for starch presence, indicating a color change as a method of observation. Spectrophotometric experiments are recommended for measuring amylase activity, with a search for relevant studies yielding multiple potential methods. Specific studies are highlighted, including those examining the inhibition of beta-amylase and kinetic determination of alpha-amylase activity, which could provide useful protocols for setting up the experiment. Participants encourage selecting a suitable method from the literature and offer assistance with any questions regarding the assays.
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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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