effect of ph on amylase enzyme (Required Practical)

Question 1

what are the steps to see the effect of ph on amylase

Answer 1

1. Place one drop of iodine solution into each spotting tile
2. take three test tubes and add each one (1 test tube will only have one type of solution): 2 cm cubes of starch solution
   2 cm cubed of amylase solution
   2 cm cubed of PH 5 buffer solution
3. put all in water bath at 30 degrees leave for 10 mins to allow solution to each correct temperature
4. Combine all 3 solutions into one and mix with stirring rod put back in water bath and start stopwatch
5. after 30 sec use string rod to transfer one drop of solution into spotting tile with the iodine (iodine will turn blue black id starch is present)
6. take a sample every 30 sec and continue until iodine stays orange
7. when iodine stays orange the reaction is complete and no starch is present and record the time
8. repeat the experiment with different ph e.g 6 7 8

Question 2

Name some problems with this experiment method and can you give some solutions

Answer 2

We only take a sample very 30 sec which means there is only approximate time for the reaction to comlete (To fix this we can take smaples every 10sec)

we are looking for the time when the iodin does no go blue black which is not obvious the colour change is gradual and some may have small amount of blue-black mixed with orange so it’s hard to see when the reaction is done

(To fix this ask several people to look at the spotting tile and decide when the reaction is complete)