VITAMINS &

COENZYMES
Co-factors
Required by enzymes in carrying out their catalytic function
Help bind the substrate to the active site
Prosthetic groups
Co-factors that are firmly/permanently bound to the protein

Co-enzymes
Small heat-stable organic molecules which can readily dissociate off
an enzyme protein and be dialyzed away, derived mostly from
vitamins

CH8: Vitamins & Co-Enzymes

Metal ion activator groups
Form coordination complexes between enzyme & substrate,
activating the latter by promoting electron shifts
Apoenzyme + co-factor Holoenzyme
(protein) (non-protein) (protein & cofactor)
Inactive Inactive Active

VITAMINS
Organic compounds that cannot be synthesized by an organism, but
are essential for the maintenance of normal metabolism
Serve as materials for the synthesis of co-enzymes

CH8: Vitamins & Co-Enzymes

Nicotinic Acid (Niacin) & Nicotinamide (Niacinamide)
Function: As nicotinic acid or nicotinamide it is a precursor for the
coenzymes NAD and NADP.
Deficiency: Pellagra (dermatitis, diarrhea, dementia and if severe, death)
Manifestations: Angular cheilosis, glossitis, ulcerative glossitis
Sources: Legumes, meat
RDA: 19 mg/day

CH8: Vitamins & Co-Enzymes

Riboflavin (Vitamin B2)
Function: Riboflavin is a precursor for the coenzymes, FMN and FAD.
Deficiency: atrophy of filliform papillae, angular cheilosis
Sources: Milk, liver, green vegetables
RDA: 1.7 mg/day

CH8: Vitamins & Co-Enzymes

Biotin
Function: Biotin acts as a coenzyme in
carboxylation reactions in covalent
linkage to lysine side chains of enzymes.
Deficiency: Dermatitis, anorexia, nausea,
muscle pain
Sources: Intestinal microorganisms, liver,
vegetables
RDA: An RDA has not been established.

CH8: Vitamins & Co-Enzymes

Thiamine (Vitamin B1)
Function: Thiamine is converted to thiamine
pyrophosphate which is a cofactor for
oxidative decarboxylation of alpha-keto acids
and for transketolase.
Deficiency: Beri-beri and Wernicke-Korsakoff
syndrome. Polyneuritis, cardiac pathology,
edema
Sources: Beans, nuts, fruits, etc.
RDA: 1.5 mg/day

CH8: Vitamins & Co-Enzymes

Pyridoxine (Vitamin B6)
Function: Pyridoxine (pyridoxol) is a
precursor of pyridoxal phosphate
which is a coenzyme for enzymes
catalyzing transamination,
deamination, decarboxylation and for
glycogen phosphorylase.
Deficiency: Dermatitis, convulsions,
microcytic anemia
Sources: Liver, fish, nuts, whole grain
cereals
RDA: 2 mg/day

CH8: Vitamins & Co-Enzymes

Folic Acid
Function: Folic acid is a precursor for
tetrahydrofolate that is used as a
carrier of one carbon units at
different levels of oxidation.
Deficiency: Megaloblastic anemia, GI
disturbances
Sources: Intestinal bacteria, liver,
yeast and green vegetables
RDA: 400 µg/day. Before food
supplementation, folic acid deficiency
was the most common vitamin
deficiency in the United States

CH8: Vitamins & Co-Enzymes

Cyanocobalamin (Vitamin B12)
Chemical Nature: Vitamin B12 is a complex multiple ring structure which includes a
cobalt atom.
Function: Vitamin B12 (cobalamin) derived cofactors are used for two important
reactions:
I. methylmalonyl CoA isomerase requires 5-deoxyadenosyl cobalamin
II. Homocysteine: tetrahydrofolate methyl transferase requires methyl cobalamin
and N5-methyltetrahydrofolate
Effect of Deficiency: Pernicious anemia and degeneration of spinal cord neurons
Sources: Meat and milk
RDA: 3 µg/day

CH8: Vitamins & Co-Enzymes

Panthotenic Acid
Function: Pantothenic acid is a
precursor of pantotheine. This
structure forms part of coenzyme A.
Deficiency: A deficiency is practically
unknown. There may be nausea and
fatigue.
Sources: Pantothenic acid is widely
distributed and is synthesized by some
intestinal bacteria
RDA: An RDA has not been established.
5-10 mg/ day might be used.

CH8: Vitamins & Co-Enzymes

Ascorbic Acid (Vitamin C)
Function: Ascorbic acid is a water soluble antioxidant and it promotes the
hydroxylation of proline residues of collagen. Ascorbic acid is also required
in the synthesis of carnitine, dopamine and bile acids.
Deficiency: Scurvy (hemorrhage, impaired wound healing and bone
formation).
Sources: Fruits and vegetables
RDA: 60 mg/day

CH8: Vitamins & Co-Enzymes

CARBOHYDRATES
 Polyhydroxycarbonyl compounds (aldehydes or ketones) or
substances that yield these compounds upon hydrolysis

ISOMERISM
Existence of 2 or more compounds with the same composition but
different constitution

A. Structural Isomers
B. Stereoisomers
1. Geometric or cis-trans isomerism
2. Optical Isomers
a. Enantiomers or optical antipodes
b. Diastereomers

CH9: Carbohydrates
 Have the same molecular formula, but different attachments

Skeletal or chain isomers Position isomers

Functional isomers

CH9: Carbohydrates
GEOMETRIC or CIS-TRANS ISOMERISM
Fumaric Acid (trans)
MP = 302°C
Insoluble in water

Maleic Acid (cis)

MP = 130°C
Soluble in water

OPTICAL ISOMERS
ENANTIOMERS OR OPTICAL ANTIPODES DIASTEREOMERS
Mirror images

Melting Point

Boiling Point

Solubility

CH9: Carbohydrates
Equimolar
amounts of
dextrorotary and
levorotary isomers
Do not exhibit
optical activity

CH9: Carbohydrates
CH9: Carbohydrates
1. All 4 sugars are D- sugars because they have the same
configuration sa D- glyceraldehyde on the penultimate carbon, C-
5.
2. D- fructose is a keto hexose and is a structural isomer of other 3.
3. The 3 aldohexoses are not mirror images of each other, hence
they are diastereomers.
4. There are 4 chiral carbons, hence there are 16 expected optical
isomers, i.e., 8 pairs of enantiomers.
5. D(+) glucose and D(+) mannose are epimers at C-2.
6. D(+) glucose and D(+) galactose are epimers at C-4.

Apparently, a special type of diastereomerism exists in:

a. Epimers – differ in a single carbon atom
b. Anomers – differ in a single atom, and the single atom
is carbon-1. CH9: Carbohydrates
• Skeletal or Structural Functions
• Storage
• Nutrient
• Reservoir of chemical energy, to be enlarged or
depleted as the organism needs

CH9: Carbohydrates
According to the number of sugar units
Monosaccharides – simple sugars
Could not be hydrolyzed further into
smaller units under reasonably mild
conditions
Soluble in water

Oligosaccharides
Hydrolyzable polymers of
monosaccharides containing 2-6
molecules of simple sugar

CH9: Carbohydrates
According to the number of sugar units
Polysaccharides
Very long chains or polymers of
monosaccharides
May be linear or branched in structure
Usually tasteless
Water insoluble
A. Homopolysaccharides – made up
of only 1 kind of monosaccharide
B. Heteropolysaccharides – made up
of 2 or more different kinds of
monosaccharides
Hyaluronic acid

CH9: Carbohydrates
According to functional groups
Aldose Ketose
contain the aldehyde group contain the ketone group

CH9: Carbohydrates
According to number of carbon atoms
Triose Pentose

Tetrose Hexose

CH9: Carbohydrates
• Simple aldehydes undergo the Kiliani-Fischer
synthesis easily.
• Aldohexoes undergo this reaction with difficulty.
• Glucose & other aldoses fail to give a positive
Schiff's test (magenta color with Schiff's reagent)
given by most aldehydes.
• Glucose is relatively inert to O2 whereas aldehydes
are notoriously auto-oxidizable.
• Glucose exhibits mutarotation

CH9: Carbohydrates
Puckered chair and boat forms:

CH9: Carbohydrates
Fischer projection formulas

Haworth formulas

CH9: Carbohydrates
• Monosaccharides exhibit mutarotation (form α and β
anomers)

• When these sugars act as reducing agents, they are

in turn oxidized
Mild OAs aldonic acids
(C-1 → COOH)

Glucose strong OAs, e.g., HNO3 aldaric acids

(CHO & C-6 → COOH)

Enzymes uronic acids

(C-6 → COOH) CH9: Carbohydrates
• The aldehyde and ketone functions may be reduced
to sugar alcohols

H2Pt or NaBH4 fermentation

Glucose D-sorbitol sorbose ascorbic acid
aerobacter
suboxydans

• They form glycosides

CH3OH, HCl
Methyl β-D glucopyranoside

Glucose CH3I or
(CH3)2SO4 Penta-O-methyl 2,3,4,6 tetra-O- methyl D- glucose

Another monosaccharide
Disaccharide
CH9: Carbohydrates
Monosaccharides give characteristic color reactions
in several qualitative tests.
This reaction is the basis for the Molisch test for
carbohydrates

CH9: Carbohydrates
Molisch Test
Napththol, then heat with concetrated
Reagents & Procedure H2SO4, general test for carbohydrates
orcinol, 30% HCl FeCl3 solution

Product Responsible Furfural and

for Color hydroxy furfural

Positive Result Violet ring

CH9: Carbohydrates
Bial's Orcinol Test

Presence of pentoses & nucleotides

Reagents & Procedure with pentoses

Product Responsible Formation of furfural (pentoses) &

for Color hydroxyfurfural (hexoses) which yields bl
ue green compound in the presence of
Fe+3 ions

Positive Result Trioses, uronic acid,

heptoses give blue

CH9: Carbohydrates
Seliwanoff's Test

Reagents & Procedure HCL + resorcinol differentiates

aldohexoses from ketohexoses

Product Responsible Dehydrated ketoses react faster to

for Color give colored condensation products

Bright cherry red

Positive Result with ketohexose,
aldohexose give
pink color

CH9: Carbohydrates
Benedict's Test

Reagents & Procedure CuSO4, Na2CO3, sodium citrate

Product Responsible Cu2O

for Color

Positive Result Brick red precipitate

CH9: Carbohydrates
Fehling's Test

Reagents & Procedure CuSO4, Na2CO3, sodium citrate

Product Responsible Cu2O

for Color

Positive Result Brick red precipitate

CH9: Carbohydrates
Barfoed's Test

Reagents & Procedure Cu (Ac)2 in dilute HCl, can distinguish between

reducing mono- and disaccharides
Product Responsible Cu2O
for Color

Positive Result Brick red precipitate

CH9: Carbohydrates
Tollen's Test

Reagents & Procedure AgNO3, NH3

Product Responsible Ag mirror

for Color

Positive Result Deposition of Ag mirror

CH9: Carbohydrates
Phenylhydrazine Test

Reagents & Procedure Phenylhydrazine + heat

Product Responsible phenylosazone

for Color

Positive Result Yellow precipitate

CH9: Carbohydrates
MALTOSE
-consists of 2 glucose units linked α 1,4, obtained as intermediates in
starch hydrolysis by the amylases
-it has a free anomeric carbon, and can exist in a and b forms (can
mutarotate, hence it is a reducing sugar

CH9: Carbohydrates
CELLOBIOSE
-has 2 glucose units linked β 1,4.
-it is formed by the hydrolysis of cellulose
-can mutarotate
-a reducing sugar

CH9: Carbohydrates
ISOMALTOSE
-has 2 glucose units linked α -1,6.

CH9: Carbohydrates
LACTOSE
-has 1 glucose and 1 galactose unit linked β -1,4.
-it is the disaccharide in milk.
-can mutarotate and is reducing.

CH9: Carbohydrates
 STORAGE POLYSACCHARIDES
A. STARCH
- the storage polysaccharide of higher plants.
-consists of 2 fractions.

1. Amylose - contains D glucose units linked α–1,4 with a reducing and

non-reducing end.
Molecular weight = few thousand to about 150,000.
With iodine = BLUE
(due to the ability of the halogen to occupy a position in the interior of
the helical coil of glucose units when amylose is suspended in water)

CH9: Carbohydrates
 STORAGE POLYSACCHARIDES
A. STARCH
2. Amylopectin – a branched polysaccharide.
Short chains (about 30 units) of glucose linked α–1,4 and α–1,6.
Molecular weight = 500,000 or higher
With iodine = PURPLE to RED

CH9: Carbohydrates
 STORAGE POLYSACCHARIDES
A. STARCH
Starch is susceptible to the action of the following hydrolytic enzymes:
α-amylase found in the digestive tract of animals (saliva & pancreatic juice)
β-amylase found in plants

When α– and β– amylase attack amylopectin, α-1,4 near branchpoints and

a-1,6 linkages are not hydrolyzed, leaving a highly branched core called
"limit dextrin"

Debranching enzyme or α–1,6 glucosidase hydrolyzes bonds at branch

points yielding a mixture of glucose and maltose.

CH9: Carbohydrates
 STORAGE POLYSACCHARIDES
A. STARCH
Starch is susceptible to the action of the following hydrolytic enzymes:
α-amylase found in the digestive tract of animals (saliva & pancreatic juice)
β-amylase found in plants

When α– and β– amylase attack amylopectin, α-1,4 near branchpoints and

a-1,6 linkages are not hydrolyzed, leaving a highly branched core called
"limit dextrin"

Debranching enzyme or α–1,6 glucosidase hydrolyzes bonds at branch

points yielding a mixture of glucose and maltose.

CH9: Carbohydrates
 STORAGE POLYSACCHARIDES
B. GLYCOGEN
-structure is similar to amylopectin but more highly branched (branch
points every 8-10 glucose) and when acted upon by α– and β– amylases
give glucose + maltose + "limit dextrin"

NUTRIENT POLYSACCHARIDES – e.g., insulin, found in bulbs of

many plants, consists fructofuranose units joined together by b-2,1
glycosidic linkage.

CH9: Carbohydrates
 STRUCTURAL POLYSACCHARIDES
A. CELLULOSE
-consists of D glucose units linked β–1,4.
-found in cell walls of plants.
Strong mineral acids cause partial hydrolysis

B. PECTINS
-contain arabinose, galactose and galacturonic acid.

C. HEMICELLULOSES
-are not cellulose derivatives
-a homopolymer of D-xylose linked β–1,4.

D. CHITIN
-a homopolymer of N-acetyl D glucosamine
-found in the shell of crustaceans and scales of insects.
CH9: Carbohydrates