Amylase & pH PRESENTATION

Which pH does Amylase work best at?

In this experiment we will be looking at the effects of pH on the enzyme Amylase

METHOD

Step 01

Step 02

Step 03

Label 5 boiling tubes 6, 7, 8, 9 and AIdentify all equipment to be used, and ensure your workspace is tidy, set-up and organised!

Place tube 6 and tube A into the bath for 2 minutes, while they are warming, place 1 drop of iodine into each well of the spotting tile - DON'T WRITE ON THE SPOTTING TILE.

Using a measuring cylinder, add 5ml of starch solution to each of the tubes 6, 7, 8 and 9.Using a pipette, place 2ml of amylase into tube A

METHOD

Step 04

Step 05

Step 06

In a clean measuring cylinder, measure out 5ml of the required pH buffer. Remove the 2 tubes from the water bath. Get ready with the stopwatch!

Start the stop watch and empty the remainder of the pipette solution back into tube 6. After 30 seconds extract some more solution and put one drop in the next spotting well, empty the rest back into tube 6. Repeat until the iodine no longer chnages colour

Add the 5ml of pH Buffer to the pH 6 tube containing the starch solution.Add the amylase solution and immediately extract a drop using a clean pipette and place a drop into the FIRST spot of the spotting tile.

SET UP

GET EVERYTHING READY

• Split into Pairs or Three's
• Double-check you have everything you need - including paper, pen/pencil, ruler, goggles and gloves

Safety Precautions -

• Wear goggles as some buffers are harmful
• Avoid getting enzymes or iodine on skin or clothes as they can cause irritation and/or staining - wash off with water in case of contact

Method

Step 07

Step 08

Step 09

Repeat with the other pH buffers, 7 and 8, and if possible 9

Clean up - your equipment, your workspace and wash your hands!

Record your results in the relevant tables.Then draw a graph for the results of table 2 - with pH on the x-axis and time on the y-axis

Consider

What caused the results observed?

Why?

What?

Which pH?

What?

What does this mean for enzymes in the body e.g. the stomach?

What effect do the other pH buffers have?

What does "denaturing" mean?

Which pH produced the fastest reaction?

Method Summary

Add 5ml of starch solution to each tube labelled 6,7,8 & 9

Label your boiling tubes

Add 2ml of amylase to tube labelled A

Place both tubes into a water bath for 2 minutes

Put 1 drop of iodine into Each well of the spotting tile

Add 2ml of amylase to Tube labelled A

Prepare 5ml of the required pH buffer And add to the starch solution And then add the amylase.

Immediately extract a small amount of the mixture and Drop it into the first well

Repeat every 30s until no colour change occurs Note the time at which this happens

Repeat the experiment with the other tubes!

Recap

What have you learned?

• What is an enzyme?
• Why are they important?
• What is unique about them during a reaction?
• What is the difference between a substrate and a product?
• What does amylase react with?
• What does Lipase reacts with?
• What do proteases react with?
• Name 2 things that can affect how effective an enzyme is?
• Where in the body are enzymes produced?
• What are enzymes made of?