1. Biological Macromolecules & enzyme
Heba Al-Tamimi 

2. Macromolecules Examples: 1. Carbohydrates 2. Lipids 3. Proteins
4. Nucleic acids
All of the above are polymers except lipids
Polymers : formed by joining one or more building blocks called MONOMERS.

3. Dehydration Synthesis
Polymers are formed by dehydration synthesis that involves removal of water. HO H
H2O HO H

4. Hydrolysis
Separates monomers by “adding water” HO H
H2O HO H

5. Carbohydrates

6. Carbohydrates: Structure
Classfication
Monosaccharides: simple sugars (monosaccharides)
Ex.: glucose
Polymer (chain of units): complex carbohydrates (or polysaccharides)
Ex.: starch, cellulose, chitin, glycogen
Disaccharide: 2 simple sugars bonded together

7. Carbohydrates Monosaccharide: one sugar unit
Examples: glucose (C6H12O6)
Fructose
Galactose
glucose

8. Carbohydrates Disaccharide: double sugar Examples:
Sucrose (glucose+fructose)
Lactose (glucose+galactose)
Maltose (glucose+glucose)
glucose

9. Carbohydrates
Polysaccharides: polymers composed of many sugar building blocks
Examples: starch
glycogen
cellulose
glucose
cellulose

10. Carbohydrates: Indicator Tests
Reducing sugars :
Benedict’s test
is used as a simple test for  detecting reducing sugars
Complex Sugars(Polysaccharides)
Lugol’s test /Iodine

11. 5-1 Benedict’s solution + Cu2O 1- + CuSO4/0H Maltose 2- reducing
-CooH
+ Cu2O
(Red –brown precipitate) 1-
+ CuSO4/0H
Maltose 2-
reducing
Glucose + Galucose

12. Benedict’s solution Sucrose Non-reducing Lactose Non-reducing 3- 4-
Glucose + Fructose
Lactose 4-
Non-reducing
Glucose + Galactose

13. Benedict’s test
starch
glucose
maltose
sucrose
H2O
2cm
Benedict
1cm solution
Fill 5 of test tubes to I cm with following solution: {glucose, maltose, sucrose, starch & water}
Fill each of the test tube to the 3 cm mark with Benedict’s reagent
Place test tube in hot water bath (5 min)
Observe color change

14. 5-2 The iodine test(lugol’s)for starch
Detect the presence of starch
Starch :glucose polymer
Glycogen: glucose polymer
Iodine potassium iodide, I2KI, interacts w coiled starch and becomes bluish black color
So ,form is important

15. iodine test(lugol’s)
glucogen
2 drop
Lugol’s
glucose
sucrose
starch
H2O
2cm solution
Fill 5 of test tubes to I cm with following solution: {glucose, sucrose, starch ,glucogen & water}
Add 2 drops of Iodine
Observe color change

16. Positive Test
Negative Test

17. Proteins

18. Proteins: Structure Proteins are made up of… Monomer (basic unit):
amino acids
20 different kinds!
Amino acids linked by peptide bonds

19. Proteins Ninhydrin test for amino acids (aas.)
aas. having free amino groups are readily detected with ninhydrin -> purple or violet
The Biuret test for polypeptide
The amino group in one aas. is linked to carboxyl group on an adjacent aas. by a peptide bond
this band is the site of action for the Biuret test for protein , Cu 2+must complex with at least four to six peptide bounds to produce violet color

20. 5-3 Ninhydrin test for amino acid
a.a
5 drop ninhydrin
albumin
glycogen
milk
H2O
+ heat
2 cm solution
Fill 5 of test tubes to I cm with following solution: {albumin ,glycogen, milk ,a.a (glycine) & water}
Add drops of ninhydrin reagent
Place test tube in hot water bath (5 min)
Observe color change
@ sink Skin irritation

21. The Biuret test for polypeptide
a.a
albumin
glycogen
milk
H2O
2 cm Biuret
2 cm solution
Fill 5 of test tubes to 3 cm with following solution: {albumin ,glycogen, milk ,a.a (glycine) & water}
Fill each test tube to the 4 cm with Biuret solution and mix
Observe color change

22. 5-4 The Biuret test for polypeptide
1% CuSO4 -> Cu2+ Complexes w at least 4-6 peptide bonds and produces violet color

23. Lipids

24. Lipids: Structure
Fat is constructed from 2 kinds of smaller molecules :
1-glycerol
2-fatty acid
Lipids are not soluble in water, but soluble in organic solvents.

25. Lipids “demo” 5-6 Lipid Solubility test
Lipids are insoluble in polar solvent and soluble in nonpolar solvent
fill 2 tubes to the 2 cm mark with corn oil
In First tube, add 5 ml of tap water
In second tube, add 5 ml of mineral oil
Detect solubility
5-5 Sudan red for fat (book)
Sudan red is a lipid soluble dye
Add 10 drops of Sudan red to tube number 1 in above experiment
Observe color , where is the red color found?

26. Nucleic Acids

27. Nucleic Acids: Structure
Nucleic Acids are made up of…
Monomers (basic units): nucleotides
Two types:
Deoxyribonucleic acid (DNA-double helix)
Ribonucleic acid (RNA-single strand)

28. 5-9 Dische diphenylamine test “demo”
In this test ,acidic condition convert deoxyribose to a molecule that binds with diphenylamine to form a blue complex.
Number 3 test tube 1-3
Into tube 1,place10 drops of banana DNA
Into tube 2,place 10 drops of standard DNA
Into tube 3,place 10 drops of distilled water
Add 1 ml of diphenylamine regent
Place in a boiling water bath for 10 min
Transfer the tubes to ice
Observe color change.
Banana DNA
Standard DNA
water

29. Enzymes…

30. Enzyme
Substrate(s) +enzyme product(s) We will take 2 examples 1- casein (protein milk)+Rennin coagulation 2- catechol +catechol oxidase benzoquinone
Optimal conditions
Optimal condition
Optimal condition

31. Enzyme (Rennin) Rennin is found in the stomach of young mammals
The enzyme coagulate the casein ,a protein found in milk
Optimal condition
pH = ~7 (neutral)
Temperate = 37 C

32. 6-1 Effect of temperature on enzyme activity
Put 3 ml Rennin
Place at 100 C for 5 min
Place at 0 C for 5 min
Place at 37 C for 5 min
Add milk (2/3) full ,then incubate to 37 C for 15 min
“Accurate measures”

33. 6-2 Effect of pH on enzyme activity
Put 3 ml Rennin
Add 3 ml HCl
Add 3 ml NaOH
Add 3 ml H2O
Add milk to each tube (2/3) full ,then incubate at 37 C for 15 min

34. 6-4 Enzyme (Catechol oxidase) “demo”
Catechole oxidase is an enzyme that turns bruised or cut fruits and vegetables brown
Catechol is a clear chemical ,when it reacts with oxygen in presence of the enzyme catechol oxidase ,it forms the dark brown product ,benzoquinone

35. Catechol oxidase “Demo”
Prepare 3 test tubes using the following table

36. Go To Work……