Scribd
Upload
10 views

Practical Enzyme

Uploaded by

M.Adil
Copyright
© © All Rights Reserved
Available Formats
Download as DOCX, PDF, TXT or read online on Scribd
10 views

Practical Enzyme

Uploaded by

M.Adil
Copyright
© © All Rights Reserved
Available Formats
Download as DOCX, PDF, TXT or read online on Scribd
You are on page 1/ 3

Invertase

Introduction

Our most common food sugar—the disaccharide, sucrose—is formed in all green plants. The

metabolism of sucrose in the animal body begins with the action of invertase (sucrase) which

hydrolyzes the disaccharide to two monosaccharides, fructose and glucose. This same enzyme is also

produced by plants and fungi.

Materials

Sucrose (0.25 M solution)

Glucose (0.25 M solution)

Benedicts solution (in dropper bottle)

Distilled water (100 ml)

Hot plate with water bath

Stirring rods (2)

Beaker (50 ml)

Test tubes (5)

Test tube rack

Pipet or syringe to measure 3 ml

Tes-Tape (tape from drug store used to test for glucose in urine samples)

Balance or teaspoon

Procedure

1. Prepare yeast by mixing 1 teaspoon (3 g) dry yeast with 20 ml of distilled water. Let stand for 20

minutes.

2. Fill each of two test tubes 1/3 full with sucrose solution.

3. Add 3 ml yeast suspension to one tube (label 1). Mix.

4. Add 3 ml distilled water to the other tube. Mix.

5. After 10 minutes test each test tube plus the yeast suspension with a strip of Tes-Tape. (Benedict's

test for reducing sugars can also be used here since sucrose will give a negative Benedict's test and

glucose/fructose will give a positive test (yellow-orange-red) depending on the amount of reducing

sugar present. Remove about 2 ml of the solution, place in another test tube, add 10 drops of
Benedict's solution and place in a boiling water bath for about 3 minutes. Prepare a glucose

solution for comparison.)

Results

For best results, do not go over the recommended incubation period. Both the Tes-Tape and

Benedict's give positive tests for samples from tube 1 while the tube without yeast gives negative

results.

Amylase

Introduction

Amylase is an enzyme that catalyses the hydrolysis of the polysaccharide starch (amylose) to the

disaccharide maltose. It is readily abundant in saliva, but somewhat unpleasant to obtain in large

quantities. It is widely distributed in plant tissues, but is most abundant in seeds, where it apparently

functions in initiating the breakdown of stored starch to glucose which is needed in large amounts

during germination.

Materials tarch-agar plates (0.2% soluble starch, 2% agar)

Distilled water (in wash bottle)

Procedure

1. Prepare starch-agar plates (do not have to be sterile if used within a day or two). Allow to solidify

and cool.

2. Use a wax pencil to label the bottom of the plate: “soaked seeds”, “boiled seeds”, “dry seeds”, etc.

(You might want to include a few drops of saliva from your mouth for comparison.)

3. Use a sharp razor blade to cut the corn grain longitudinally and place, cut surface down, onto the

agar surface. (You may wish to dissect out the embryo.) Be sure to space corn grains at least 2

cm from each other.

4. Incubate for 30 minutes.

5. Remove corn and rinse plate gently with distilled water.

6. Flood plate with iodine solution, swish around as color develops, rinse with distilled water, record

results. (Any clear areas of agar can be removed and tested for sugars.)

Results
After flooding the plates with iodine solution, the agar will stain a deep purple in all areas where

starch remains. Areas of agar where dead seeds were placed will be purple, likewise for dry seeds

(unless the incubation period is much longer) since dormant seeds produce very little amylase. Areas

of the agar covered by saliva, or by a living embryo, will appear clear since the starch has been broken

down.

You might also like