Effect of pH on enzyme activity- amylase

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Discussion Overview

The discussion centers around the investigation of the effect of pH on the enzyme amylase, focusing on experimental design, methods for measuring enzyme activity, and obtaining quantitative results. Participants explore various approaches to conducting the experiment and measuring outcomes.

Discussion Character

  • Exploratory
  • Technical explanation
  • Homework-related

Main Points Raised

  • One participant seeks guidance on how to carry out an investigation into the effect of pH on amylase activity, specifically requesting equipment and methods for obtaining quantitative results.
  • Another participant suggests measuring amylase activity by determining the rate of sugar production from starch using different pH buffers.
  • A different approach is proposed involving a 1% starch solution with amylase and various pH buffers, followed by testing for reducing sugars using Benedict's reagent, although the participant expresses uncertainty about obtaining quantitative results.
  • One participant recommends measuring the intensity of the color change with Benedict's reagent to quantify results, suggesting the use of dilution and a set of standards for comparison.
  • Another participant raises concerns about the complexity of the Benedict's test and proposes an alternative method of measuring the remaining starch using iodine, suggesting timing the hydrolysis process and observing the color change.

Areas of Agreement / Disagreement

Participants present multiple competing views on how to measure enzyme activity and obtain quantitative results, with no consensus reached on a single method or approach.

Contextual Notes

Participants express uncertainty about the best methods for quantifying results and the potential challenges associated with different testing methods, such as the need for heating in the Benedict's test and the timing of starch hydrolysis.

Who May Find This Useful

This discussion may be useful for students or individuals interested in enzyme activity experiments, particularly those exploring the effects of pH on amylase and seeking experimental design ideas.

GaMeR4LyF
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Hi guys, I've got to do a project for school on the investigation of the effect of pH on the enzyme amylase. I am abit lost right now only just started A levels. Can someone suggest how i would carry out this investigation? Equipment and method would be much appreciated! And the results need to be quantitative and not qualitative. so how can i adapt the experiment so i get quantitative results?
 
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First you need to determine how to measure the activity of the amylase. Amylase converts starch to sugar, so you will need a method to determine the sugar. Find a soluble starch and place it into different pH buffers with the amylase and measure the rate of sugar production.
 
hmm...how about i leave a 1percent starch solution with amylase with buffers of pH 2, 3, 5, 7 and 9 for 5 minutes then use benedicts reagent to test for reducing sugars... those tested positive with reducing sugars would sugest there is enzyme activity... But i don't know how to obtain quantitative results from this...
 
You would have to measure the intensity of the color change with the Benedicts reagent (used in excess) to quantify it. Dilution after the color development stage might be required. You can make up a set of standards to compare against and bracket the concentration in that way. Your eye would be the detector. You would measure how much the sample faded from blue to colorless in that example.
Alternatively, you could measure using urine test strips that diabetics use. Should cost roughly a dime per test.
 
benedict's test can be quite hectic wen you have to heat in boiling water for two minutes each time.

i'd rather test for the remaining starch. you should measure the time for the complete hydrolysis to occur. fo rthat you use a stopwatch, and at say 2 min, intervals(it really depends on the concentration of solution you use), use a dropper and place it on a spot tile, where you previously had placed iodine in the holes.

wen reaction is over, all starch would have been digested, and you will get a brown solution with the iodine. (iodine + starch gives a blue black ppt.)
 

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