Introductory Biochemistry

Chapter 2:
Carbohydrates

By: Dr. Sobia Mushtaq

Carbohydrates
Polyhydroxy aldehydes/ketones. Because of presence of OH-group.

Carbohydrates

i. Have strong interactions with water

ii. Give them functional groups to which various substituents can be added
iii. They provide possibility of strong intra and inter chain reactions via H2 bonds

Widely distributed in plants and animals and play major role in

i. Metabolism ii. Structure

Plants: glucose is synthesized from CO2 and H2O by photosynthesis and

stored in form of starch or converted to cellulose of plant framework

Animals: can synthesize some carbohydrate from fat and protein……bulk of

carbohydrate is derived from plant
Glucose

Major fuel of the tissues of mammals and universal fuel of fetus

 Converted to other carbohydrates having highly specific functions:

i. Glycogen for storage

ii. Ribose in nucleic acids
iii. And in combination with proteins such as glycoprteins and protoglycans

Diseases associated with CHOs

 Diabetes mellitus
 Glycogen storage disease
 Lactose intolerance
 Galactosemia
Classification
have the following basic composition:
I
(C H 2O )n or H - C - OH
I
 Monosaccharides - simple sugars with multiple OH groups
(aldoses/ketoses). Based on number of carbons (3, 4, 5, 6), a
monosaccharide is a triose, tetrose, pentose or hexose. Eg
glyceraldehyde
 Disaccharides - 2 monosaccharides covalently linked.
 Oligosaccharides - a few monosaccharides covalently linked.
 Polysaccharides – Most carbohydrates in nature exist as high
molecular weight polymers. And composed of simple/derived
sugars joined through glycosidic linkage. Eg starch, glycogen
 Monosaccharides
Aldoses (e.g., glucose) Ketoses (e.g., fructose)
have an aldehyde group at have a keto group,
one end. usually at C2.

H O CH 2 OH
C
C O
H C OH
HO C H
HO C H
H C OH
H C OH
H C OH
H C OH

C H 2O H
CH 2 OH

D -glucose D -fructose
 D vs L Designation
D & L designations
CHO CHO
are based on the
configuration about H C OH HO C H
the single CH2OH CH2OH
asymmetric C in D-glyceraldehyde L-glyceraldehyde
glyceraldehyde.
CHO CHO
The lower
H C OH HO C H
representations are
Fischer Projections. CH2OH CH2OH
D-glyceraldehyde L-glyceraldehyde
 Hemiacetal & hemiketal
formation
H H
An aldehyde can
react with an alcohol C O + R' OH R' O C OH
to form a hemiacetal.
R R
A ketone can react aldehyde alcohol hemiacetal
with an alcohol to
form a hemiketal.
R R

C O + "R OH "R O C OH

R' R'
ketone alcohol hemiketal
 1
CHO
Pentoses and
hexoses can cyclize H C OH
2
as the ketone or HO C H D-glucose
3
aldehyde reacts
H C OH (linear form)
with a distal OH. 4
H C OH
Glucose forms an 5
intra-molecular 6
CH2OH
hemiacetal, as the
6 CH2OH 6 CH2OH
C1 aldehyde & C5
5 5
OH react, to form a H O H H O OH
6-member pyranose H H
4 H 1 4 H 1
OH OH
ring, named after H
OH OH OH
pyran. 3 2 3 2
H OH H OH
-D-glucose -D-glucose

These representations of the cyclic sugars are

called Haworth projections.
 Sugar derivatives
COOH CHO
CH2OH
H C OH H C OH
H C OH
HO C H HO C H
H C OH
H C OH H C OH
H C OH
H C OH H C OH
CH2OH
CH2OH COOH
D-ribitol
D-gluconic acid D-glucuronic acid

 sugar alcohol - lacks an aldehyde or ketone; e.g., ribitol.

 sugar acid - the aldehyde at C1, or OH at C6, is oxidized to
a carboxylic acid; e.g., gluconic acid, glucuronic acid.
 Sugar derivatives
CH2OH CH2OH

H O H H O H
H H
OH H OH H

OH OH OH O OH
H NH2 H N C CH3
H
-D-glucosamine -D-N-acetylglucosamine

amino sugar - an amino group substitutes for a

hydroxyl. An example is glucosamine.
The amino group may be acetylated, as in
N-acetylglucosamine.
 6CH
O
2 H 6CH
O
2 H
Disaccharides: 5 5
H O H H O H
Maltose, a cleavage H H
product of starch (e.g., 4 O
H H 1 4
O
H H 1

amylose), is a O
H O O
H
2 2
disaccharide with an 3 3

a(1® 4) glycosidic link H O

H m
alto
se H O
H
between C1 - C4 OH of 2
glucoses. 6CH
O
2 H 6CH
OH2
It is the a anomer (C1 O 5 O 5 O O
H
H H
points down). H H
4
O
H H 1 O 4
O
H H 1

O
H H H
3 2 3 2

H O
H H O
H
c
ello
bio
se

Cellobiose, a product of cellulose breakdown, is the otherwise

equivalent b anomer (O on C1 points up).
The b(1® 4) glycosidic linkage is represented as a zig-zag, but
one glucose is actually flipped over relative to the other.
Other disaccharides include:
 Sucrose, common table sugar, has a glycosidic bond linking
the anomeric hydroxyls of glucose & fructose.
Because the configuration at the anomeric C of glucose is a
(O points down from ring), the linkage is a(12).
The full name of sucrose is a-D-glucopyranosyl-(12)-b-D-
fructopyranose.)
 Lactose, milk sugar, is composed of galactose & glucose,
with b(14) linkage from the anomeric OH of galactose. Its
full name is b-D-galactopyranosyl-(1 4)-a-D-
glucopyranose
Polysaccharides:
Plants store glucose as amylose or amylopectin, glucose
polymers collectively called starch.
Glucose storage in polymeric form minimizes osmotic
effects.
Amylose is a glucose polymer with a(14) linkages.
Amylopectin is a glucose polymer with mainly a(14)
linkages, but it also has branches formed by a(16) linkages.
The branches produce a compact structure & provide multiple
chain ends at which enzymatic cleavage can occur.
 CH2OH 6CH OH CH2OH CH2OH CH2OH
2
O 5 O H O H O H H O H
H H H H H
H H H H H
OH H 1 4 OH H 1 OH H OH H OH H
O O O O OH
OH 2
3
H OH H OH H OH H OH H OH
amylose

CH 2 OH CH 2 OH
H O H H O H am ylopectin
H H
OH H OH H 1
O
OH
O
H OH H OH

CH 2 OH CH 2 OH 6 CH 2 CH 2 OH CH 2 OH
H O H H O H H 5 O H H O H H O H
H H H H H
OH H OH H OH H 1 4 OH H OH H
4 O O
O O OH
OH 2
3
H OH H OH H OH H OH H OH
 CH2OH CH2OH
H O O
glycogen
H H H
H H
OH H OH H 1
O
OH
O
H OH H OH

CH2OH CH2OH 6 CH2 CH2OH CH2OH

H O H H O H H 5 O H H O H H O H
H H H H H
OH H OH H OH H 1 4 OH H OH H
4 O O
O O OH
OH
3 2
H OH H OH H OH H OH H OH

Glycogen, the glucose storage polymer in animals, is similar in structure to

amylopectin.
But glycogen has more a(16) branches.
The highly branched structure permits rapid glucose release from glycogen
stores, e.g., in muscle during exercise.
The ability to rapidly mobilize glucose is more essential to animals than to
plants.

(Amylopectin branched every 24-30 glucose residues while glycogen

branched every 4-8 glucose residues)
 CH2OH 6CH OH CH2OH CH2OH CH2OH
2
O 5 O O H O H O OH
H H H
H H H H H
OH H 1 O 4 OH H 1 O OH H O OH H O OH H
OH H H H
H 2 H
3
H OH H OH H OH H OH H OH
cellulose
Cellulose, a major constituent of plant cell walls, consists of long linear
chains of glucose with b(1®4) linkages.
This promotes intra-chain and inter-chain H-bonds and van der
Waals interactions, that cause cellulose chains to be straight &
rigid, and pack with a crystalline arrangement in thick bundles -
microfibrils.
These microfibrils are very strong.
The role of cellulose is to impart strength and
rigidity to plant cell walls, which can
withstand high hydrostatic pressure
gradients. Osmotic swelling is prevented.
Schematic of arrangement of
cellulose chains in a microfibril.
 heparan sulfate
core glycosaminoglycan
protein

transmembrane
-helix
cytosol

Proteoglycans are glycosaminoglycans that are covalently

linked to serine residues of specific core proteins. 
The glycosaminoglycan chain is synthesized by sequential
addition of sugar residues to the core protein.
 NAN NAN NAN

Gal Gal Gal

NAG NAG NAG

Man Man N-linked oligosaccharide

Man Key:
NAN = N-acetylneuraminate
NAG Gal = galactose
NAG = N-acetylglucosamine
NAG Fuc
Man = mannose
Fuc = fucose
Asn

Additional monosaccharides are added, and the N-

linked oligosaccharide chain is modified by removal
and addition of residues, to yield a characteristic
branched structure.
Glycolysis
 Glycolytic Pathways
Having covered the basic structural features of carbohydrates….Sugar
metabolism…produces energy and C-compounds to be used in
biosynthesis.

Generally regarded as a primitive process because

i. Occurs in cytosol rather than being compartmentalized into
specific organelles within eukaryotic cells
ii. Also it probably arose early in biological history before the evident
of eukaryotic organisms.

Greek roots glycos….sweet and lysis…..splitting ( So splitting of sugar

which is sweet)
 6 CH OPO 2
2 3
5 O
H H
H
4 H 1
OH
OH OH
3 2
H OH
glucose-6-phosphate

Glycolysis takes place in the cytosol of cells.

Glucose enters the Glycolysis pathway by conversion to glucose-
6-phosphate.
Initially there is energy input corresponding to cleavage of two ~P
bonds of ATP.
 6 CH2OH 6 CH OPO 2
2 3
ATP ADP
5 O 5 O
H H H H
H H
4 1 4 H 1
OH H OH
Mg2+
OH OH OH OH
3 2 3 2
H OH Hexokinase H OH
glucose glucose-6-phosphate

1. Hexokinase catalyzes:
Glucose + ATP  glucose-6-P + ADP
The reaction involves nucleophilic attack of the C6 hydroxyl O of glucose
on P of the terminal phosphate of ATP.
ATP binds to the enzyme as a complex with Mg++.
Mg++ interacts with negatively charged phosphate oxygen atoms,
providing charge compensation & promoting a favorable conformation of
ATP at the active site of the Hexokinase enzyme.
 6 CH OPO 2
2 3
5 6 CH OPO 2 1CH2OH
H O H 2 3
O
H
4 H 1 5 H HO 2
OH
OH OH H 4 3 OH
3 2
OH H
H OH
Phosphoglucose Isomerase
glucose-6-phosphate fructose-6-phosphate

2. Phosphoglucose Isomerase catalyzes:

glucose-6-P (aldose)  fructose-6-P (ketose)
The mechanism involves acid/base catalysis, with ring opening,
isomerization via an enediolate intermediate, and then ring closure.
 Phosphofructokinase
6 CH OPO 2 1CH2OH 6 CH OPO 2 1CH2OPO32
2 3 2 3
O ATP ADP O
5 H HO 2 5 H HO 2

H 4 3 OH Mg2+ H 4 3 OH
OH H OH H
fructose-6-phosphate fructose-1,6-bisphosphate

3. Phosphofructokinase catalyzes:
fructose-6-P + ATP  fructose-1,6-bisP + ADP
This highly spontaneous reaction has a mechanism similar to that of
Hexokinase.
The Phosphofructokinase reaction is the rate-limiting step of
Glycolysis.
The enzyme is highly regulated.
 2
1CH2OPO3

2C O
H O
2
HO 3C H Aldolase 3
CH2 OPO 3 1C

H 4C OH 2C O + H 2C OH
2
H C OH 1CH2OH 3 CH2OPO3
5
2 dihydroxyacetone glyceraldehyde-3-
6 CH2 OPO 3
phosphate phosphate
fructose-1,6-
bisphosphate
Triosephosphate Isomerase

4. Aldolase catalyzes: fructose-1,6-bisphosphate 

dihydroxyacetone-P + glyceraldehyde-3-P
The reaction is an aldol cleavage, the reverse of an aldol
condensation.
Note that C atoms are renumbered in products of Aldolase.
 2
1CH2OPO3

2C O
H O
2
HO 3C H Aldolase 3
CH2 OPO 3 1C

H 4C OH 2C O + H 2C OH
2
H C OH 1CH2OH 3 CH2OPO3
5
2 dihydroxyacetone glyceraldehyde-3-
6 CH2 OPO 3
phosphate phosphate
fructose-1,6-
bisphosphate
Triosephosphate Isomerase

5. Triose Phosphate Isomerase (TIM) catalyzes:

dihydroxyacetone-P  glyceraldehyde-3-P
Glycolysis continues from glyceraldehyde-3-P.
 G lycerald eh yd e -3 -p h o sp h ate
D eh yd ro gen ase
H O + H+ O O PO 3 2 
1C NAD+ NADH 1 C
+ Pi
H C OH H C OH
2 2
2 2
3 CH 2 O PO 3 3 CH 2 O PO 3

glyc erald eh yd e - 1 ,3 -b isp h o sp h o -

3 -p h o sp h a te glyc era te

6. Glyceraldehyde-3-phosphate Dehydrogenase catalyzes:

glyceraldehyde-3-P + NAD+ + Pi 
1,3-bisphosphoglycerate + NADH + H+
Exergonic oxidation of the aldehyde in glyceraldehyde- 3-phosphate, to a
carboxylic acid, drives formation of an acyl phosphate, a "high energy"
bond (~P). This is the only step in Glycolysis in which NAD+ is reduced to
NADH.
 Phosphoglycerate Kinase
O OPO32 ADP ATP O O
1C 1
C
H 2C OH H 2C OH
2+
2 Mg 2
3 CH2OPO3 3 CH2OPO3

1,3-bisphospho- 3-phosphoglycerate
glycerate
7. Phosphoglycerate Kinase catalyzes:
1,3-bisphosphoglycerate + ADP 
3-phosphoglycerate + ATP
This phosphate transfer is reversible (low DG), since one ~P bond
is cleaved & another synthesized.
 Phosphoglycerate Mutase
O O O O
C
1
C
1
H 2C OH H 2C OPO32
2
3 CH 2 OPO 3 3 CH2OH
3-phosphoglycerate 2-phosphoglycerate

8. Phosphoglycerate Mutase catalyzes:

3-phosphoglycerate  2-phosphoglycerate

Phosphate is shifted from the OH on

C3 to the OH on C2.
 Enolase
O  H 
O  OH O
O O O
C C 1
C
1
H 2 C OPO32 C OPO32 2C OPO32

3 CH2OH CH2OH 3 CH2

2-phosphoglycerate enolate intermediate phosphoenolpyruvate

9. Enolase catalyzes:
2-phosphoglycerate  phosphoenolpyruvate + H2O
This dehydration reaction is Mg++-dependent.
2 Mg++ ions interact with oxygen atoms of the substrate carboxyl group at the active
site.
The Mg++ ions help to stabilize the enolate anion intermediate that forms when a Lys
extracts H+ from C #2.
 Pyruvate Kinase
O O O O
ADP ATP
1
C 1
C

2
C OPO32 2
C O

3 CH2 3 CH3
phosphoenolpyruvate pyruvate

10. Pyruvate Kinase catalyzes:

phosphoenolpyruvate + ADP  pyruvate + ATP
This phosphate transfer from PEP to ADP is
spontaneous.
Balance sheet for ~P bonds of ATP:
 2 ATP expended
 4 ATP produced (2 from each of two 3C fragments
from glucose)
 Net production of 2 ~P bonds of ATP per glucose.
Glycolysis - total pathway, omitting H+:
glucose + 2 NAD+ + 2 ADP + 2 Pi 
2 pyruvate + 2 NADH + 2 ATP
In aerobic organisms:
 Pyruvate produced in Glycolysis is oxidized to CO2
via Krebs Cycle
 NADH produced in Glycolysis & Krebs Cycle is
reoxidized via the respiratory chain, with production
of much additional ATP. 
 G lyceraldehyde-3-phosphate
D ehydrogenase
H O + H+ O O PO 3 2
1C NAD+ NADH 1C
Fermentation: + Pi
H C OH H C OH
2 2
Anaerobic CH 2 O PO 3
2
CH 2 O PO 3
2
3 3
organisms lack a
respiratory chain. glyceraldehyde- 1,3 -bisphospho-
3-phosphate glycerate

They must reoxidize NADH produced in Glycolysis through some

other reaction, because NAD+ is needed for the Glyceraldehyde-3-
phosphate Dehydrogenase reaction.
Usually NADH is reoxidized as pyruvate is converted to a more
reduced compound.
The complete pathway, including Glycolysis and the reoxidation of
NADH, is called fermentation.
Fermentation Pyruvate’s Fate

• Features of fermentation pathways

– Pyruvic acid is reduced to form
reduced organic acids or
alcohols.
– The final electron acceptor is a
reduced derivative of pyruvic
acid
– NADH is oxidized to form NAD:
Essential for continued operation
of the glycolytic pathways.
– O2 is not required.
– No additional ATP are made.
– Gases (CO2 and/or H2) may be
released
 L a c ta te D e h y d ro g e n a s e
O O O O
C NADH + H+ NAD+ C
C O HC OH
CH 3 CH 3
p y ru v a te la c ta te
E.g., Lactate Dehydrogenase catalyzes reduction of the keto in pyruvate
to a hydroxyl, yielding lactate, as NADH is oxidized to NAD+.
 Lactate, in addition to being an end-product of fermentation, serves as a
mobile form of nutrient energy, & possibly as a signal molecule in
mammalian organisms.
 Cell membranes contain carrier proteins that facilitate transport of lactate.
 Skeletal muscles ferment glucose to lactate during exercise, when the
exertion is brief and intense.
 Lactate released to the blood may be taken up by other tissues, or by
skeletal muscle after exercise, and converted via Lactate Dehydrogenase
back to pyruvate, which may be oxidized in Krebs Cycle or (in liver)
converted to back to glucose via gluconeogenesis
 Pyruvate Alcohol
Decarboxylase Dehydrogenase
O O
CO2 H NADH + H+ NAD+ H
C O
C H C OH
C O
CH3 CH3
CH3
pyruvate acetaldehyde ethanol

Some anaerobic organisms metabolize

pyruvate to ethanol, which is excreted as a
waste product.
NADH is converted to NAD+ in the reaction
catalyzed by Alcohol Dehydrogenase.
Glycolysis, omitting H+:
glucose + 2 NAD+ + 2 ADP + 2 Pi 
2 pyruvate + 2 NADH + 2 ATP
Fermentation, from glucose to lactate:
glucose + 2 ADP + 2 Pi  2 lactate + 2 ATP

Anaerobic catabolism of glucose yields only 2

“high energy” bonds of ATP.
 DGo' DG
Glycolysis Enzyme/Reaction kJ/mol kJ/mol
Hexokinase -20.9 -27.2
Phosphoglucose Isomerase +2.2 -1.4
Phosphofructokinase -17.2 -25.9
Aldolase +22.8 -5.9
Triosephosphate Isomerase +7.9 negative
Glyceraldehyde-3-P Dehydrogenase -16.7 -1.1
& Phosphoglycerate Kinase
Phosphoglycerate Mutase +4.7 -0.6
Enolase -3.2 -2.4
Pyruvate Kinase -23.0 -13.9

*Values in this table from D. Voet & J. G. Voet (2004) Biochemistry, 3 rd

Edition, John Wiley & Sons, New York, p. 613.
Gluconeogenesis
Glycolysis and gluconeogenesis constitute oppositely
directed conversions with different ATP-ADP
stoichiometries.
Several features of Glycolysis and gluconeogenesis systems
indicate that regulation of these conversions are of special
importance to organism
In mammalian liver, massive glycogen synthesis takes place
after a meal. If meal was high in protein, much of glucose
that is incorporated into glycogen is produced from
pyruvate and oxaloacetate via gluconeogenesis.
During fasting much or all of the glycogen…..glucose 6-PO4
and then metabolized to pyruvate.
Summary of Gluconeogenesis Pathway:
Gluconeogenesis enzyme names in red.
Glycolysis enzyme names in blue.
 Glucose-6-phosphatase
6 CH OPO 2 CH2OH
2 3
5 O O
H H H H
H H2O H
4
OH H 1
OH H + Pi
OH OH OH OH
3 2
H OH H OH
glucose-6-phosphate glucose

Hexokinase or Glucokinase (Glycolysis) catalyzes:

glucose + ATP  glucose-6-phosphate + ADP
Glucose-6-Phosphatase (Gluconeogenesis) catalyzes:
glucose-6-phosphate + H2O  glucose + Pi
Glucose-6-phosphatase enzyme is embedded in the endoplasmic
reticulum (ER) membrane in liver cells.
 Phosphofructokinase 
6 CH OPO 2 1CH2OH 6 CH OPO 2 1CH2OPO32
2 3 2 3
O ATP ADP O
5 H HO 2 5 H HO 2

H 4 3 OH H 4 3 OH
Pi H2O
OH H OH H
fructose-6-phosphate fructose-1,6-bisphosphate
 Fructose-1,6-biosphosphatase

Phosphofructokinase (Glycolysis) catalyzes:

fructose-6-P + ATP  fructose-1,6-bisP + ADP
Fructose-1,6-bisphosphatase (Gluconeogenesis) catalyzes:
fructose-1,6-bisP + H2O  fructose-6-P + Pi
 Pyruvate Carboxylase PEP Carboxykinase
O O
C
O O O O
C ATP ADP + Pi C O GTP GDP C
C O CH2 C OPO32
HCO3 C CO2
CH3 CH2
O O
pyruvate oxaloacetate PEP

Bypass of Pyruvate Kinase (2 enzymes):

Pyruvate Carboxylase (Gluconeogenesis) catalyzes:
pyruvate + HCO3- + ATP  oxaloacetate + ADP + Pi
PEP Carboxykinase (Gluconeogenesis) catalyzes:
oxaloacetate + GTP  PEP + GDP + CO2
Global Control in liver cells includes reciprocal
effects of a cyclic AMP cascade, triggered by the
hormone glucagon when blood glucose is low.
Phosphorylation of enzymes & regulatory proteins
in liver by Protein Kinase A (cAMP Dependent
Protein Kinase) results in
 inhibition of glycolysis
 stimulation of gluconeogenesis,
making glucose available for release to the blood.
1. Action:-
Glycolysis is the breakdown of glucose up to formation of pyruvate (in aerobic conditions) or
lactate (in anaerobic conditions). While gluconeogenesis is the synthesis of glucose from
other then carbohydrates substances such as pyruvate, alpha ketoglutarate, gluconeogenic
amino acids, lactate and gluconeogenic glycerol.
2. Site of pathway:
Site of pathway of glycolysis is liver and muscles. While glyconeogenesis occurs in liver and
kidneys.
3. Site inside cell:
First step of glycolysis occurs in cytoplasm and generates only a small amount of energy
while the rest of steps occur in mitochondria. While the first three reactions of glycogenolysis
occurs in mitochondria and the rest in cytosol.
4. Types:
Glycogenesis has two types i.e aerobic glycolysis in which glucose is reduced up to pyruvate
which then enters kreb’s cycle and is further reduced. And another type is anaerobic
glycolysis in which pyruvate is converted into lactate.
While gluconeogenesis has not any sub-types
5. ATP Production:
ATPs are produced in glycognesis while no ATPs are produced in gluconeogeneis.
Gluconeogenesis only produce glucose at the end of pathway
6. Rate limiting Steps:
There are three rate limiting and irreversible steps in glycolysis. The enzymes involved in
these three rate limiting steps i.e rate limiting enzymes are: hexokinase/glucokinase,
phosphofructokinase and pyruvate kinase.
While there are four rate limiting steps in gluconeogenesis. The rate limiting enzymes are :
pyruvate carboxylase, phosphoenol pyruvate carboxykinase, fructose 1, 2 bisphophatase,
and glucose 6 phosphate phosphatase.
 Kreb’s cycle
OR
TCA (Tricarboxylic acid cycle)
OR
Citric acid cycle
 Stoichiometry of the Citric Acid Cycle
 Two carbon atoms enter
the cycle in the form of
acetyl CoA.
 Two carbon atoms leave
the cycle in the form of
CO2 .
 Four pairs of hydrogen
atoms leave the cycle in
four oxidation reactions
(three molecules of NAD+
one molecule of FAD are
reduced).
 One molecule of GTP,
is formed.
 Two molecules of water
9 ATP
 are (2.5 ATP per NADH, and
consumed.
1.5 ATP per FADH2) are produced
during oxidative phosphorylation.
 1 ATP is directly formed in the
citric acid cycle.
 1 acetyl CoA generates
approximately 10 molecules of
ATP.
 An Overview of the Citric Acid Cycle
A four-carbon oxaloacetate condenses with
a two-carbon acetyl unit to yield a six-
carbon citrate.
An isomer of citrate is oxidatively
decarboxylated and five-carbon -
ketoglutarate is formed.
-ketoglutarate is oxidatively
decarboxylated to yield a four-carbon
succinate.
Oxaloacetate is then regenerated from
succinate.
Two carbon atoms (acetyl CoA) enter the
cycle and two carbon atoms leave the cycle
in the form of two molecules of carbon
dioxide.
Three hydride ions (six electrons) are
transferred to three molecules of NAD+, The function of the citric acid
one pair of hydrogen atoms (two electrons) cycle is the harvesting of high-
is transferred to one molecule of FAD. energy electrons from acetyl CoA.
 1. Citrate Synthase
• Citrate formed from acetyl CoA and oxaloacetate
• Only cycle reaction with C-C bond formation
• Addition of C2 unit (acetyl) to the keto double
bond of C4 acid, oxaloacetate, to produce C6
compound, citrate

citrate synthase
 2. Aconitase
• Elimination of H2O from citrate to form C=C
bond of cis-aconitate
• Stereospecific addition of H2O to cis-aconitate
to form isocitrate

aconitase aconitase
 3. Isocitrate Dehydrogenase
• Oxidative decarboxylation of isocitrate to
a-ketoglutarate (a metabolically irreversible reaction)
• One of four oxidation-reduction reactions of the cycle
• Hydride ion from the C-2 of isocitrate is transferred to NAD + to form
NADH
• Oxalosuccinate is decarboxylated to a-ketoglutarate

isocitrate dehydrogenase isocitrate dehydrogenase

4. The -Ketoglutarate Dehydrogenase Complex
• Similar to pyruvate dehydrogenase complex
• Same coenzymes, identical mechanisms
E1 - a-ketoglutarate dehydrogenase (with TPP)
E2 – dihydrolipoyl succinyltransferase (with
flexible lipoamide prosthetic group)
E3 - dihydrolipoyl
dehydrogenase (with FAD)

a-ketoglutarate
dehydrogenase
 5. Succinyl-CoA Synthetase
• Free energy in thioester bond of succinyl CoA is
conserved as GTP or ATP in higher animals (or
ATP in plants, some bacteria)
• Substrate level phosphorylation reaction

+ HS-
Succinyl-CoA
Synthetase

GTP + ADP GDP + ATP

6. The Succinate Dehydrogenase Complex
• Complex of several polypeptides, an FAD prosthetic group
and iron-sulfur clusters
• Embedded in the inner mitochondrial membrane
• Electrons are transferred from succinate to FAD and then
to ubiquinone (Q) in electron transport chain
• Dehydrogenation is stereospecific; only the trans isomer is
formed

Succinate
Dehydrogenase
 7. Fumarase
• Stereospecific trans addition of water to the
double bond of fumarate to form L-malate
• Only the L isomer of malate is formed

Fumarase
 8. Malate Dehydrogenase
Malate is oxidized to form oxaloacetate.

Malate
Dehydrogenase
 Functions of the Citric Acid Cycle
• Integration of metabolism. The citric acid cycle
is amphibolic (both catabolic and anabolic).
The cycle is involved in Intermediates of the
the aerobic catabolism cycle are starting points
of carbohydrates, for many anabolic
lipids and amino acids. reactions.

• Yields energy in the form of GTP (ATP).

• Yields reducing power in the form of NADH2

and FADH2.
 Synthesis of
glycogen Glucose
Pentose phosphate
pathway

Glycogen Glucose-6- Ribose, NADPH

phosphate

Degradation of
glycogen

Glycolysis Gluconeogenesis

Ethanol Pyruvate Lactate

Fatty Acids Acetyl Co A Amino Acids

The citric acid

cycle is the
Most fuel
final common
pathway for the molecules
oxidation of fuel enter the
molecules — cycle as
amino acids, acetyl
fatty acids, and coenzyme A.
carbohydrates.
Photosynthesis: Calvin cycle
 Photosynthesis
• Overview of Photosynthesis
– Light-dependent Reactions:
• Light energy is harvested by photosynthetic pigments and
transferred to special reaction center (photosystem)
chlorophyll molecules.
• The light energy is used to strip electrons from an electron
donor (the electron donor goes from a reduced to an
oxidized state).
• The electrons are shuttled through a series of electron
carriers from high energy state to a low energy state.
• During this process, ATP is formed.
• In the cyclic pathway of electron transport, electrons are
returned to the electron transport chain
• In the non-cyclic pathway, the electrons are used to reduce
NAD (or NADP) to NADH (or NADPH)
 Photosynthesis
b)Light-independent Reactions:
• ATP and NADH (NADPH) from the light-
dependent reactions are used to reduce CO2 to
form organic carbon compounds (carbon
fixation).
• The reduced organic carbon is usually
converted into glucose or other carbohydrates.
The Calvin cycle has three phases
– Carbon fixation
– Reduction (energy input, reducing equiv input)
– Regeneration of the CO2 acceptor
(energy input – “priming step”)

65
 Carbon
Fixation
Acid -COOH
Input of
energy

Reduction
Rubisco
Priming
Step

Aldehyde
-C=O
H

Regenerate
What started 66
with
Integration of Light-Dependent
and
Light-Independent Reactions

They generally occur AT THE SAME TIME

Light reaction Calvin cycle
H2O CO2

Light
NADP+
ADP
+P1
RuBP 3-Phosphoglycerate
Photosystem II
Electron transport chain
Photosystem I
ATP G3P
NADPH Starch
(storage)
Figure 10.21 Amino acids
Fatty acids 67
Chloroplast
O2 Sucrose (export)
Summary
1. Photosynthetic “light” reactions produce
ATP and reducing potential NADPH + H+

2. Dark reactions use ATP and reducing potential

to synthesize carbohydrates
- powers reduction of 3-carbon acid
to 3-carbon aldehyde
- powers regeneration of starting material
5-carbon di-phosphate (priming step for CO2 fixation)

3. Rubisco enzyme regulated tightly by allosteric modulators

pH, and reducing status of stroma

4. O2 interferes with carbon fixation by Rubisco enzyme

5. Metabolic “tricks” to avoid photorespiration

- C4 metabolism - CAM metabolism

68