Effect of pH on enzyme activity- amylase

In summary,The author is looking for advice on how to carry out an investigation into the effect of pH on the enzyme amylase. They are interested in either measuring the activity of the enzyme or finding a way to quantify it. They are also interested in finding a way to determine when the reaction has completed.
  • #1
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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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  • #2
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.
 
  • #3
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...
 
  • #4
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.
 
  • #5
benedict's test can be quite hectic wen you hav 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.)
 

1. What is the role of pH in enzyme activity?

The pH level of a solution can greatly affect the activity of enzymes, including amylase. Enzymes have an optimal pH range in which they function most efficiently. Deviations from this range can result in a decrease in enzyme activity or complete denaturation.

2. How does pH affect the structure of amylase?

Changes in pH can disrupt the hydrogen bonds and ionic bonds that maintain the structure of amylase. This can lead to changes in the shape of the enzyme, which can affect its ability to bind to substrates and catalyze reactions.

3. What is the optimal pH for amylase activity?

The optimal pH for amylase varies depending on the source of the enzyme. However, it typically falls within the acidic range, around pH 6.0-7.0. This is because amylase is found in the stomach, which has an acidic pH.

4. How does pH affect the rate of enzyme activity?

The rate of enzyme activity is directly affected by the pH level. As the pH deviates from the optimal range, the rate of enzyme activity decreases. This is due to changes in the enzyme's structure and its ability to bind to substrates and catalyze reactions.

5. Can extreme pH levels completely inhibit amylase activity?

Yes, extreme pH levels can completely inhibit amylase activity. If the pH is too high or too low, the enzyme can become denatured and lose its ability to function. This is why maintaining the optimal pH range is crucial for enzyme activity.

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