T.Y. B.Sc.

BC-504
Unit-I
Carbohydrate Metabolism
Definition of metabolism:
Metabolism is defined as the sum of all the biochemical reactions catalysed by enzymes taking place
in a cell or organism. It includes following two processes:
a. Catabolism: It is break down of complex molecules (carbohydrates, proteins, fats etc) into
smaller, simpler end products (lactic acid, CO2, NH3 etc). In this process energy is released in
the form of ATP, NADH, NADPH and FADH2.
b. Anabolism: It is also called biosynthesis. Small, simple precursors are built up into larger and
more complex molecules like carbohydrate, proteins, fats etc. Anabolic process requires
energy in the form of ATP, NADH, NADPH and FADH 2.

Glycolysis:
In glycolysis, one molecule of glucose is degraded in a series of enzyme-catalyzed reactions to yield
two molecules of the three-carbon compound pyruvate. During glycolysis free energy released is
conserved in the form of ATP and NADH. The most common type of glycolysis is the Embden–
Meyerhof–Parnas (EMP pathway), which was discovered by Gustav Embden, Otto Meyerhof, and
Jakub Karol Parnas.
Glycolysis occurs in ten steps. The first five steps come under preparatory phase. Next five steps
come under payoff phase in which energy is released.
Steps are as follows:

A) Preparatory phase of glycolysis:

a) Phosphorylation of Glucose
Glucose is phosphorylated at C-6 to yield glucose 6-phosphate. ATP act as the phosphate group
donor. This reaction is catalyzed by hexokinase in presence of Mg2+.

6 CH2OH ADP 6 CH2OPO3

ATP
H 5C O H H 5C O H
H Mg2+ H
4C C1 4C H C1
OH H OH
Hexokinase
OH C C OH OH C C OH
3 2 3 2
H OH H OH
Glucose Glucose-6-phosphate

b) Conversion of Glucose 6-Phosphate to Fructose 6-Phosphate

The enzyme phosphohexose isomerase catalyzes the reversible isomerization of glucose 6-
phosphate to fructose-6-phosphate.

1
 6 CH2OPO3
6 CH2OPO3 1 CH2OH
H 5C O H O
H 2+
Mg
4C OH H C1 5C H OH C2
Phosphohexose isomerase
OH C C OH C C
3 2 H 4 3 OH
H OH H
OH
Glucose-6-phosphate
Fructose-6-phosphate

c) Phosphorylation of Fructose 6-Phosphate to Fructose 1, 6-Bisphosphate

Phosphofructokinase-1 (PFK-1) catalyzes the transfer of a phosphate group from ATP to fructose 6-
phosphate to yield fructose 1,6-bisphosphate.

6 CH2OPO3 1 CH2OH ATP ADP 6 CH2OPO3 1 CH2OPO3

O O
5C H OH C2 Mg2+ 5C H OH C2
Phosphofructokinase-1
C C C C
H 4 3 H 4 3
OH OH
OH H H
OH
Fructose-6-phosphate Fructose-1,6-bisphosphate

d) Cleavage of Fructose 1,6-Bisphosphate

The enzyme fructose 1,6-bisphosphate aldolase, catalyzes a reversible aldol condensation. Fructose
1,6-bisphosphate is cleaved to yield two different triose phosphates, glyceraldehyde-3-phosphate,
and dihydroxyacetone phosphate.

6 CH2OPO3 1 CH2OPO3 CHO

CH2OPO3
O
5C H
Aldolase CHOH
OH C 2 C O +
C C CH2OH CH2OPO3
H 4 3 OH Dihydroxyacetone phosphate Glyceraldehyde 3-phosphate
OH H
Fructose-1,6-bisphosphate

e) Interconversion of the Triose Phosphates

Dihydroxyacetone phosphate, is rapidly and reversibly converted to glyceraldehyde-3-phosphate by
triose phosphate isomerase.

CH2OPO3 CHO
Triose phosphate isomerase
C O CHOH

CH2OH CH2OPO3
Dihydroxyacetone phosphate Glyceraldehyde 3-phosphate

This reaction completes the preparatory phase of glycolysis.

B) Payoff Phase of Glycolysis:

One molecule of glucose yields two molecules of glyceraldehyde 3-phosphate.

f) Oxidation of Glyceraldehyde 3-Phosphate to 1,3-Bisphosphoglycerate

Oxidation of glyceraldehyde 3-phosphate to 1,3-bisphosphoglycerate is catalyzed by glyceraldehyde-
3-phosphate dehydrogenase using inorganic phosphate. The hydrogen acceptor is NAD+ yielding the
NADH.

2
 O

O H O O P O
C O C O
Glyceraldehyde 3-phosphate
2 CHOH + 2 HO P O 2 CHOH
O
dehydrogenase
CH2OPO3 Inorganic phosphate 2 NAD 2 NADH + H CH2OPO3
Glyceraldehyde 3-phosphate 1,3-bisphosphoglycerate

g) Phosphoryl Transfer from 1,3-Bisphosphoglycerate to ADP

The enzyme phosphoglycerate kinase transfers the high-energy phosphate group from the carboxyl
group of 1,3-bisphosphoglycerate to ADP, forming ATP and 3-phosphoglycerate.

O O P O O O
C O C
Phosphoglycerate kinase
2 CHOH + 2 ADP 2 CHOH + 2 ATP
Mg2+
CH2OPO3 CH2OPO3
1,3-bisphosphoglycerate 3-phosphoglycerate

h) Conversion of 3-Phosphoglycerate to 2-Phosphoglycerate

The enzyme phosphoglycerate mutase catalyzes a reversible shift of the phosphoryl group between
C-2 and C-3 of glycerate; Mg 2+ is essential for this reaction.

O O O O
C C
Phosphoglycerate mutase
2 CHOH 2 CHOPO3
Mg2+
CH2OPO3 CH2OH
3-phosphoglycerate 2-phosphoglycerate

i) Dehydration of 2-Phosphoglycerate to Phosphoenolpyruvate

Enolase promotes reversible removal of a molecule of water from 2-phosphoglycerate to yield
phosphoenolpyruvate (PEP).

O O O O
C C
Enolase
2 CHOPO3 2 C OPO3 + 2 H2O

CH2OH CH2
2-phosphoglycerate Phosphoenolpyruvate

j) Transfer of the Phosphoryl Group from Phosphoenolpyruvate to ADP

The last step in glycolysis is the transfer of the phosphoryl group from phosphoenolpyruvate to ADP,
catalyzed by pyruvate kinase, which requires K+ and either Mg2+ or Mn2- to produce pyruvic acid.

O O O O
C C
Pyruvate kinase
2 C OPO3 + 2 ADP 2 C O + 2 ATP
Mg2+ & K+ CH3
CH2
Phosphoenolpyruvate Pyruvate
The Overall Balance Sheet of glycolysis:

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The left-hand side of the following equation shows all the inputs and the right-hand side shows all
the outputs (each molecule of glucose yields two molecules of pyruvate):

Glucose + 2ATP + 2NAD+ + 4ADP + 2Pi 2Pyruvate + 2ADP + 2NADH + 2H+ + 4ATP + 2H2O

Cancelling out common terms on both sides of the equation gives the overall equation for glycolysis
under aerobic conditions:

Glucose + 2NAD+ + 2ADP + 2Pi 2Pyruvate + 2NADH + 2H + + 2ATP + 2H2O

Bioenergetics of glycolysis:
In preparatory phase of glycolysis two ATP molecules are utilised while in payoff phase 4 ATP and
two NADH molecules are directly formed. The two molecules of NADH under aerobic condition are
reoxidized to NAD+ by transfer of their electrons to the electron transfer chain to produce ATP
molecules.
Reaction ATP/NADH ATP formed
formed/utilized
Glucose Glucose 6-phosphate -1 ATP -1
Fructose 6-phosphate Fructose 1,6-bisphosphate -1 ATP -1
2 (Glyceraldehyde 3-phosphate) 2 (1,3-bisphosphoglycerate) 2 NADH 5
2 (1,3-bisphosphoglycerate) 2 (3-phosphoglycerate) 2ATP 2
2 (phosphoenolpyruvate) 2 (Pyruvate) 2 ATP 2
Total 7
Note: 1 NADH = 2.5 ATP; Negative sign indicates ATP utilization

4
 Glucose
ATP
Hexokinase Mg2+
ADP

Glucose 6-phosphate
Phosphohexose isomerase

Fructose 6-phosphate
ATP
Phosphofructokinase-1 Mg2+
ADP

Fructose 1,6-bisphosphate

Aldoase

Dihydroxyacetone phosphate + Glyceraldehyde 3-phosphate x 2

2NAD+ + 2Pi Glyceraldehyde 3-phosphate
Triose phosphate isomerase dehydrogenase
2NADH + 2 H+

1,3- bisphosphoglycerate x 2
2ADP
2+
Mg Phosphoglycerate kinase
2ATP

3-phosphoglycerate x 2

Mg2+ Phosphoglycerate mutase

2-phosphoglycerate x 2

2H2O Enolase
Phosphoenolpyruvate x 2
2ADP
Mg2+ Pyruvate kinase
2ATP

Pyruvate x 2

Fate of Pyruvate
 NADH is formed from NAD+ during glycolysis.
 The redox balance of the cell has to be maintained for further cycles of glycolysis to
continue.
 NAD+ can be regenerated by one of the following reactions /pathways:
o Pyruvate is converted to lactate
o Pyruvate is converted to ethanol
o In the presence of O2, NAD+ is regenerated by Electron Transport Chain (ETC).
Pyruvate is converted to acetyl-CoA which enters TCA cycle and gets completely
oxidized to CO2.

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Lactate Fermentation
 Formation of lactate catalyzed by lactate dehydrogenase:
CH3-CO-COOH + NADH + H+ ↔ CH3-CHOH-COOH + NAD+
 In highly active muscle, there is anaerobic glycolysis because the supply of O 2 cannot keep
up with the demand for ATP.
 Lactate builds up causing a drop in pH which inactivates glycolytic enzymes. End result is
energy deprivation and cell death; the symptoms being pain and fatigue of the muscle.
 Lactate is transported to the liver where it can be reconverted to pyruvate by the LDH
reverse reaction
Ethanol fermentation
 Formation of ethanol is catalyzed by 2 enzymes
 Pyruvate decarboxylase catalyses the first irreversible reaction to form acetaldehyde:
CH3-CO-COOH → CH3-CHO + CO2
 Acetaldehyde is reduced by alcohol dehydrogenase. It is a reversible reaction:
CH3-CHO + NADH + H+ ↔ CH3CH2OH + NAD+
 Ethanol fermentation is used during wine-making

Oxidative Decarboxylation of Pyruvate to Acetyl-CoA (Activated Acetate):

Before entering the citric acid cycle, pyruvate is oxidised to acetyl-CoA by the pyruvate
dehydrogenase (PDH) complex.
The PDH complex is composed of multiple copies of three enzymes: pyruvate dehydrogenase, E1
(with its bound coenzyme TPP); dihydrolipoyl transacetylase, E2 (with its covalently bound lipoyl
group); and dihydrolipoyl dehydrogenase, E3 (with its coenzymes FAD and NAD).
O O
CoASH NAD NADH
C
TPP, Lipoate, FAD O S CoA
C O + CO2
C
CH3 Pyruvate dehydrogenase complex E1, E2, E3
CH3
Pyruvate Acetyl CoA
 In step 1 pyruvate reacts with the bound thiamine pyrophosphate (TPP) of pyruvate
dehydrogenase (E1). Pyruvate undergoes decarboxylation to produce hydroxyethyl
derivative.
CO2

E1-TPP + CH3COCOOH E1-TPP-CHOH-CH3

Pyruvate dehydrogenase Pyruvate Hydroxyethyl derivative

 In step 2, pyruvate dehydrogenase transfers acetyl group from TPP to the oxidized form of
the lipoyllysyl group of dihydrolipoyl transacetylase (E2), to form the acetyl thioester of the
reduced lipoyl group.

E1-TPP-CHOH-CH3 + E2
E1-TPP + E2
Hydroxyethyl derivative Pyruvate dehydrogenase
S S
Dihydrolipoyl transacetylase S HS
C CH3

O
Acetyl thioester of lipoyl group

 Step 3 is a transesterification in which the -SH group of CoA replaces the -SH group of E2 to
yield acetyl-CoA and the fully reduced (dithiol) form of the lipoyl group.

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 E2 + CoA-SH E2 + CH3CO-S-CoA
CoA Acetyl-CoA
S HS SH HS
C CH3 Fully reduced dithiol form of lipoyl group

O
Acetyl thioester of lipoyl group

 In step 4 dihydrolipoyl dehydrogenase (E3) promotes transfer of two hydrogen atoms from
the reduced lipoyl groups of E2 to the FAD of E3, restoring the oxidized form of the
lipoyllysyl group of E2.

E2 + E3-FAD E2 + E3-FADH2
Dihydrolipoyl dehydrogenase
SH HS S S
Fully reduced dithiol Dihydrolipoyl transacetylase
form of lipoyl group

 In step 5 the reduced FADH2 of E3 transfers a hydride ion to NAD, forming NADH. The
enzyme complex is now ready for another catalytic cycle.
E3-FADH2 + NAD E3-FAD + NADH + H
Dihydrolipoyl dehydrogenase

Bioenergetics:
In glycolysis two pyruvate molecules are formed. So ATP generation calculated accordingly.

Reaction ATP/NADH formed/utilized ATP ultimately formed

2 Pyruvate 2 Acetyl-CoA + 2CO2 2NADH 5
Note: 1 NADH = 2.5 ATP

Krebs Cycle / Tricarboxylic Acid Cycle (TCA) / Citric Acid Cycle:

Acetyl-CoA produced by oxidative decarboxylation of pyruvate enter first cyclic pathway citric acid
cycle (first product of the pathway is citric acid), also called the tricarboxylic acid (TCA) cycle (citric
acid contains 3 –COOH groups) or the Krebs cycle (after its discoverer, Hans Krebs).

a) Condensation:
The first reaction of the cycle is the condensation of acetyl-CoA with oxaloacetate to form citrate,
catalyzed by citrate synthase.

H2O CoA SH H2C COO

O S CoA
C O C COO HO C COO
+ H2C COO Citrate Synthase
CH3 H2C COO
Acetyl CoA Oxaloacetate Citrate

b) Dehydration and Hydration:

The enzyme aconitase catalyzes the reversible transformation of citrate to isocitrate (by hydration),
through the intermediary formation of cis-aconitate (by dehydration).

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 H2O H2O
H2C COO H2C COO H2C COO

HO C COO C COO H C COO

Aconitase Aconitase
H2C COO HC COO HO C COO
H
Citrate cis- aconitate Iso-citrate

c) Oxidative Decarboxylation:
Isocitrate dehydrogenase catalyzes oxidative decarboxylation of isocitrate to form α-ketoglutarate
and CO2. Mn2+ is required as co-factor. NAD+ acts as hydrogen acceptor.
NADH + H CO2
H2C COO NAD H2C COO
H C COO Mn2+
H C H
HO C COO Isocitrate dehydrogenase O C COO
H alpha-ketoglutarate
Iso-citrate

d) Oxidative Decarboxylation:
α-ketoglutarate is converted to succinyl-CoA and CO2 by the action of the α-ketoglutarate
dehydrogenase complex; NAD+ serves as electron acceptor and CoA as the carrier of the succinyl
group.
CoA-SH
NAD NADH + H
H2C COO H2C COO
H C H H C H + CO2
alpha-ketoglutarate
O C COO dehydrogenase complex O C S-CoA
alpha-ketoglutarate Succinyl-CoA

e) Substrate level Phosphorylation:

The cleavage of the thioester bond of succinyl-CoA is coupled to the phosphorylation of GDP. This
reaction is catalyzed by succinyl-CoA synthetase.
CoA-SH
GDP + Pi GTP
H2C COO H2C COO
H C H H C H
O C
Succinyl CoA synthetase COO
S-CoA
Succinyl-CoA Succinate

f) Dehydrogenation:
The succinate is oxidized to fumarate by the succinate dehydrogenase. In this reaction FAD accepts
hydrogen and gets converted to FADH2.
FAD FADH2
H2C COO
HC COO
H C H
Succinate dehydrogenase OOC CH
COO
Fumarate
Succinate

g) Hydration:
The reversible hydration of fumarate to L-malate is catalyzed by fumarase. This enzyme is highly
stereospecific; it catalyzes hydration of the trans double bond of fumarate but not the cis double
bond of maleate (the cis isomer of fumarate).

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h) Dehydrogenation:
In the last reaction of the citric acid cycle, NAD-linked L-malate dehydrogenase catalyzes the
oxidation of L-malate to oxaloacetate.

Acetyl CoA + H2O CoA-SH

NADH + H+ Citrate synthase

Oxaloacetate Citrate Aconitase
NAD + H2O
Malate dehydrogenase

Malate cis-aconitate

H2O
Fumarase Aconitase
TCA Cycle
H2O
Isocitrate
NAD+
Fumarate
Isocitrate dehydrogenase
FADH2
NADH + H+ + CO2
Succinate dehydrogenase
-ketoglutarate
FAD Succinate
NAD+ + CoA-SH
Succinyl-CoA -ketoglutarate dehydrogenase complex
GTP + CoA-SH NADH + H+ + CO2
Succinyl CoA synthetase GDP + Pi

Bioenergetics:

Reaction ATP/NADH formed/ utilized ATP formed

2Isocitrate 2 alpha-ketoglutarate 2NADH 5
2 alpha-ketoglutarate 2 Succinyl-CoA 2NADH 5
2 Succinyl-CoA 2 Succinate 2GTP 2
2 Succinate 2Fumarate 2FADH2 3
2 Malate 2 Oxaloacetate 2NADH 5
Total 20
Note: 1 NADH = 2.5 ATP; 1FADH2 = 1.5ATP; 1GTP = 1ATP

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Glyoxylate Cycle:
In plants, certain invertebrates, and some microorganisms acetyl CoA formed from lipids can serve
both as an energy-rich fuel and as a source of phosphoenolpyruvate for carbohydrate synthesis. In
these organisms, enzymes of the glyoxylate cycle catalyze the net conversion of acetate to succinate,
an intermediate of the citric acid cycle:
 Acetyl-CoA condenses with oxaloacetate to form citrate, and citrate is converted to
isocitrate, exactly same as in the citric acid cycle.
 However, isocitrate is not the broken down by isocitrate dehydrogenase.
 Isocitrate is cleaved by isocitrate lyase, forming succinate and glyoxylate.
 The glyoxylate then condenses with a second molecule of acetyl-CoA to yield malate, in a
reaction catalyzed by malate synthase.
 The malate is subsequently oxidized to oxaloacetate, which can condense with another
molecule of acetyl-CoA to start another turn of the cycle.
 Each turn of the glyoxylate cycle consumes two molecules of acetyl-CoA and produces one
molecule of succinate, which is then available for biosynthetic purposes.
 The succinate may be converted through fumarate and malate into oxaloacetate, which can
then be converted to phosphoenolpyruvate by PEP carboxykinase, and thus to glucose by
gluconeogenesis.

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 O S CoA
C
CH3
Acetyl CoA

Citrate Synthase

H2C COO
O C COO
HO C COO
NADH + H H2C COO
H2C COO
Oxaloacetate
Citrate
L-Malate dehydrogenase
Aconitase
NAD
COO
Glyoxylate Cycle
HC OH
H2C COO H2C COO
L-Malate
H C COO
HO C COO
Malate synthase O
H
C O Iso-citrate
Isocitrate lyase
C O
H
O S CoA Glyoxylate
C
CH3
H2C COO
Acetyl CoA
H C H
COO
Succinate

Pentose Phosphate Pathway/ Phosphogluconate pathway/Hexose monophosphate (HMP)

pathway
The major catabolic fate of glucose-6-phosphate is glycolytic breakdown to pyruvate. In some
tissues, glucose 6-phosphate is oxidised to ribose 5-phosphate by the pentose phosphate pathway.
In this oxidative pathway, NADP+ is the hydrogen acceptor, yielding NADPH.

Functions of HMP pathway:

 Rapidly dividing cells, such as those of bone marrow, skin, and intestinal mucosa, use the
riboses to make RNA and DNA. This ribose sugar is made available by HMP pathway.
 In tissues like erythrocytes, lens and cornea, NADPH produced in HMP pathway is required
for reduction of oxygen radicals.
 For fatty acid synthesis (in liver, adipose, lactating mammary gland) or synthesis of
cholesterol and steroid hormones (in liver, adrenal gland, gonads) NADPH is required.
HMP pathway occurs in two phases- Oxidative and non-oxidative phase.

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Oxidative Phase:
1. Glucose-6-phosphate is oxidised by glucose 6-phosphate dehydrogenase (G6PD) to form 6-
phosphoglucono-δ-lactone. NADP+ is hydrogen acceptor to produce NADPH.
2. The 6-phosphoglucono-δ-lactone is hydrolyzed to 6-phosphogluconate by a lactonase.
3. 6-phosphogluconate undergoes oxidation and decarboxylation by 6-phosphogluconate
dehydrogenase to form the ketopentose ribulose-5-phosphate. This reaction generates a
second molecule of NADPH.
4. Phosphopentose isomerase converts ribulose-5-phosphate to its isomer, ribose-5-
phosphate.
Oxidative phase occurs in tissue which requires pentose sugar for synthesis of DNA and RNA. Its
overall equation is
Glucose 6-phosphate + 2NADP+ + H2O → ribose 5-phosphate + CO2 + 2NADPH + 2H+
HOHC
NADP+
H C OH 6-phosphogluconate
dehydrogenase
HO C H Mg2+
O CO2
H C OH NADPH + H+
H C
CH2OH
CH2OPO3
Glucose 6-phosphate
C O
NADP+
H C OH

Glucose 6-phosphate
dehydrogenase Mg2+ H C OH

CH2OPO3
NADPH + H+
D-Ribulose 5-phosphate
O C
Phosphopentose
H C OH isomerase

HO C H CHO
O
H C OH H C OH

H C H C OH

CH2OPO3 H C OH
6-phospho-glucono--lactone
CH2OPO3
H2O
Lactonase D-Ribose 5-phosphate
Mg2+

O C O

H C OH

HO C H

H C OH

H C OH

CH2OPO3
6-phosphogluconate

Non-oxidative Phase:
Some tissues require only NADPH. In these tissues, ribose 5-phosphates produced in the oxidative
phase are recycled into glucose-6-phosphate. In this phase there is series of rearrangements of the
carbon skeletons to produce Glucose 6-phosphate.
1. Ribulose-5-phosphate is first epimerized to xylulose-5-phosphate. The reaction is catalysed
by enzyme epimerase.

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 2. Xylulose-5-phosphate combines with Ribose 5-phosphate to produce Sedoheptulose 7-
phosphate and Glyceraldehyde-3-phosphate. This reaction is catalysed by transketolase.
3. Sedoheptulose 7-phosphate combines with Glyceraldehyde 3-phosphate to produce
Fructose 6-phosphate and Erythrose 4-phosphate. This reaction is catalysed by
transaldolase.
4. Fructose 6-phosphate is isomerised to Glucose 6-phosphate. This reaction is catalysed by
enzyme phosphohexose isomerase.
5. Two molecules of glyceraldehyde-3-phosphate are converted to a molecule of fructose 1,6-
bisphosphate by the same way as in gluconeogenesis.
6. Fructose 1,6-bisphosphatase-1 dephosphorylates Fructose 1,6-bisphosphate to Fructose 6-
phosphate.
7. Phosphohexose isomerase converts Fructose 6-bisphosphate to glucose-6-phosphate.

Ribose 5-phosphate
Phosphohexose
Phosphopentose Sedoheptulose 7-phosphate isomerase
isomerase Fructose 6-phosphate Glucose 6-phosphate

Phosphohexose
Ribulose 5-phosphate isomerase
Transketolase Transaldolase

Epimerase Glyceraldehyde 3-phosphate Erythrose 4-phosphate Fructose 6-phosphate

Xylulose 5-phosphate Reverse of glycolysis

Transketolase

Xylulose 5-phosphate Glyceraldehyde 3-phosphate

Glycogenolysis:
Glycogen structure:
Glycogen is the carbohydrate reserve in animals, hence often referred to as animal starch. It is
present in high concentration in liver, followed by muscle, brain etc. Glycogen is also found in plants
that do not possess chlorophyll (e.g. yeast and fungi).
The structure of glycogen is similar to that of amylopectin with more number of branches. Glucose is
the repeating unit in glycogen joined together by α (1→4) glycosidic bond and α (1→6) glycosidic
bond at branching points. The molecular weight (up to 1 x 108) and the number of glucose units (up
to 25,000) vary in glycogen depending on the source glycogen are obtained.

6CH2OH 6CH2OH 6CH2OH

H 5C O H H 5C O H H 5C O H
H H H
4C OH H C1 4C OH H C1 4C OH H C1

O C C O C C O C C
3 2 3 2 3 2
H OH H OH H OH alpha-(1 6) branch point
Non-reducing end
O
6 CH2OH 6CH2OH 6 CH2OH 6CH2

H 5C O H H 5C O H H 5C O H H 5C O H
H H H H
4C OH H C1 4C OH H C1 4C OH H C1 4C OH H C1
O C C O C C O C C O C C
3 2 3 2 3 2 3 2
H OH H OH H OH H OH
Non-reducing end Glycogen Reducing end

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Glycogen breakdown (Glycogenolysis):
 It is the breakdown of glycogen to form glucose-1-phosphate, which is converted to glucose-
6- phosphate.
 Glycogenolysis is not a reversal of glycogenesis, separate set of cytosolic enzymes is
required.
 Glycogenolysis is catalyzed by three enzymes, glycogen phosphorylase, debranching enzyme
and phosphoglucomtase.
Glycogen phosphorylase:
 It catalyzes removal of glucose residues from the non-reducing ends on the outer branches
of glycogen by adding inorganic phosphate and producing glucose-l-phosphate. The action of
the enzyme stops when there is only four glucose residues on each side of the branching
point.
Debranching enzyme:
a) Glucosyl transferase activity of debranching enzyme
 It catalyzes transfer of the outer three residues of glucose at a branching point to the near
non- reducing end of the glycogen molecule, leaving only one glucose residue at the
branching point.
b) Glucosidase activity of debranching enzyme
 It catalyzes the removal of the last glucose residue at the branching point by adding water
and producing free glucose. Unbranched chain is further hydrolyzed by the Glycogen
phosphorylase enzyme.
Phosphoglucomutase:
 Glucose 1-phosphate, the end product of the glycogen phosphorylase reaction, is converted
to glucose 6-phosphate by phosphoglucomutase.

Fate of glucose-6-phosphate:
In liver:
Glucose-6-phosphate is mainly converted to glucose due to the presence of Glucose-6-phosphatase.
Free glucose formed is released to the blood, as the main function of liver glycogen is to maintain
the blood glucose level especially during fasting or carbohydrate deficiency.
In muscle:

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Glucose-6-phosphate is oxidized by glycolysis to provide energy. Glucose-6-phosphate cannot be
converted to free glucose due to deficiency of Glucose-6-phosphatase.

Glycogenolysis under influence of hormones:

Glycogen phosphorylase, which degrades glycogen to glucose 1-phosphate, is under regulation of
hormone. In muscle, glycogen phosphorylase is under control of epinephrine while in liver, it is
under control of glucagon. Both hormones activate glycogen phosphorylase in same way which is as
follows:
 When in muscle and blood glucose level decreases, epinephrine and glucagon bind at
receptors present on myocyte and hepatocyte respectively.
 They activate G-protein which is present just beneath plasma membrane.
 Activated G-protein activates adenylyl cyclase which converts ATP to cAMP.
 Elevated [cAMP] initiates an enzyme cascade.
 The rise in [cAMP] activates cAMP dependent protein kinase, also called protein kinase A
(PKA).
 PKA then phosphorylates and activates phosphorylase b kinase to its active form
phosphoryase a kinase.
 Phosphorylase a kinase phosphorylates and activates glycogen phosphorylase b to glycogen
phosphoryase a, and thus stimulating glycogen breakdown.

Myocyte Hepatocyte

Epinephrine Glucagon

G-protein (inactive) G-protein (active)

Adenylyl cyclase
ATP Cyclic AMP 20 x molecules

Inactive Protein Kinase A Active Protein Kinase A 10 x molecules

Phosphorylase b kinase (inactive) Phosphorylase a kinase (active) 100 x molecules

Glycogen phosphorylase (inactive) Glycogen phosphorylase (active) 1000 x molecules

Glycogen Glucose 1-phosphate 10000 x molecules

Glucose
(in muscle) Glycolysis

Blood (in liver)

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Gluconeogenesis:
 Gluconeogenesis is the synthesis of glucose from non-carbohydrate precursors including
pyruvate, lactate, glycerol and amino acids.
 Gluconeogenesis is not reverse of glycolysis.
 Out of ten, seven enzymatic reactions of gluconeogenesis are the reverse of glycolytic
reactions.
 However, three reactions of glycolysis are essentially irreversible.
 There are substitute or bypass reactions for the irreversible steps of glycolysis.
 Glycerol enters reverse glycolysis as DHAP by the action of glycerol kinase followed by
dehydrogenase.
 Lactate is converted to pyruvate by LDH.
 Glucogenic amino acids are converted to either pyruvate or intermediate products of TCA
cycle prior to gluconeogenesis.

Glucogenic amino acids

TCA intermediate→ Pyruvate Succinyl-CoA α-Ketoglutarate Fumarate Oxaloacetate
Amino acids→ Alanine Isoleucine Arginine Phenylalanine Asparagine
Cysteine Methionine Glutamate Tyrosine Aspartate
Glycine Threonine Glutamine
Serine Valine Histidine
Threonine Proline
Tryptophan

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 Glucose
Pi
ATP
Hexokinase Mg2+ Glucose 6-phosphatase
ADP

Glucose 6-phosphate H2O

Fructose 6-phosphate Pi

ATP
Phosphofructokinase-1 Mg2+ Fructose 1,6-bisphosphatase
ADP
H2O
Fructose 1,6-bisphosphate

Dihydroxyacetone phosphate + Glyceraldehyde 3-phosphate x 2

2NAD+ + 2Pi

2NADH + 2 H+

1,3- bisphosphoglycerate x 2
2ADP
Mg2+
2ATP

3-phosphoglycerate x 2

Mg2+
2-phosphoglycerate x 2

2H2O
2GDP
Phosphoenolpyruvate x 2
PEP carboxykinase
Pyruvate kinase 2ADP
Mg2+ 2GTP
2ATP Oxaloacetate x 2

Pyruvate x 2 2ADP
Pyruvate carboxylase
2ATP

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 Phosphoenolpyruvate
Cytosolic
PEP
Carboxykinase CO2

Oxaloacetate
NADH + H+
Cytosolic
Malate
dehydrogenase
NAD+
Malate

Cytoplasm

Malate PEP

NAD+ Mitochondrial
Mitochondrial PEP CO2
Malate Carboxykinase
dehydrogenase
NADH + H+ Oxaloacetate
Oxaloacetate
Pyruvate
Pyruvate Carboxylase
Carboxylase
CO2
CO2 Pyruvate
Pyruvate

Mitochondria

Pyruvate Pyruvate
NAD+
Lactate
dehydrogenase
NADH + H+
Lactate

Bypass for Puruvate Kinase

 Pyruvate synthesized by glycolysis or from amino acid is in the mitochondria.
 Here, pyruvate is first converted to oxaloacetate by the enzyme pyruvate carboxylase. One
carbon is supplied by CO2 to form the 4-C oxaloacetate. The reaction is coupled to ATP
hydrolysis making this a ligation reaction.
 Oxaloacetate (OAA) is converted to malate as mitochondrial membrane is impermeable to
OAA.
 This reaction is catalysed by the mitochondrial malate dehydrogenase enzyme.
 Malate is transported into cytoplasm through transporter.
 In cytoplasm malate is converted back to OAA with the help of cytosolic malate
dehydrogenase.
 Oxaloacetate is converted to PEP by the enzyme PEP carboxykinase. CO2 is removed and
energy in the form of GTP is utilized.

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PEP can be converted to fructose-1,6 bisphosphate by reverse glycolysis.

Bypass for Phosphofructokinase-1

 Fructose-1,6 bisphosphatase converts Fructose 1,6-bisphosphate to Fructose 6-phosphate
since Phosphofructokinase-1 reaction is irreversible.
 This removes the phosphate group from the 1 position. However, no ATP is formed.

Further reverse glycolysis leads to formation of Glucose 6-phosphate.

Bypass for Hexokinase

 Glucose-6-phosphatase converts glucose-6-phosphate to glucose since the hexokinase
reaction is irreversible.
 This removes the phosphate group from the 1 position. However, no ATP is formed.

Net Reaction for gluconeogenesis:

 2 pyruvate + 4 ATP + 2 GTP + 2 NADH + 2 H + + 6 H2O → glucose + 2 NAD+ + 4 ADP + 2 GDP + 6
Pi.
 (Net reaction for glycolysis is: Glucose + 2NAD+ + 2 ADP + 2 Pi → 2 pyruvate + 2 ATP + 2
NADH + 2 H2O)

Glycogen synthesis (glycogenesis):

Synthesis of glycogen from glucose is known as glycogenesis. It occurs mainly in the cytosol of liver
cells and muscles. Glycogenesis requires- a primer, UDP-Glucose units, two enzymes- glycogen
synthase and branching enzyme.
A. Primer:
Glycogen synthase cannot initiate chain synthesis using free glucose. It can elongate already existing
chains of more than 4 residues of glucose (primer).
This primer is either:
 A fragment of glycogen in cells.
 In absence of glycogen fragment, a protein called glycogenin act as primer. It is auto-
glycosylated at tyrosine residue by UDP-Glucose.
Tyrosine

Glycogenin OH

UDP-Glucose

Auto-glycosylate

UDP

Glycogenin O Glucose

B. Synthesis of uridine diphosphoglucose (UDP-Glu):

 Glucose is phosphorylated to glucose 6-phosphate. This reaction is catalyzed by the enzymes
hexokinase in muscle and glucokinase in liver.
 Glucose 6-phosphate is converted to glucose 1-phosphate by phosphoglucomutase.

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  Glucose 1-phosphate is converted to UDP-glucose by the action of Glucose 1-phosphate
uridylyltransferase. This reaction is irreversible by the subsequent hydrolysis of
pyrophosphate.
Hexokinase
Glucose + ATP Glucose 6-phosphate + ADP
Glucokinase

Phosphoglucomutase
Glucose 6-phosphate Glucose 1-phosphate

Glucose 1-phosphate
Glucose 1-phosphate + UTP UDP-Glucose + PPi
Uridylyltransferase

Pyrophosphatase 2Pi
PPi + H2O

UDP-glucose is the immediate donor of glucose residues in the reaction catalyzed by glycogen
synthase.
Glycogen synthase:
 It catalyzes the transfer of the glucosyl group of UDP-Glu to the non-reducing ends of
glycogen primer.
 Glycogen synthase forms successive -1-4 glycosidic linkage.
 This produces elongation of the branches of glycogen primer up to a minimum of 11 glucose
residues.
Glycogen Branching enzyme:
 Glycogen synthase cannot make the (α1→6) bonds at the branch points of glycogen.
 (α1→6) bonds are formed by the glycogen-branching enzyme, also called amylo (α1→4) to
(α1→6) transglycosylase or glycosyl-(4→6)-transferase.
 The glycogen-branching enzyme catalyses transfer of a terminal fragment of 6 or 7 glucose
residues from the non-reducing end of a glycogen to the C-6 hydroxyl group of a glucose
residue at a more interior position of the same or another glycogen chain, thus creating a
new branch.
Further glucose residues may be added to the new branch by glycogen synthase.
The biological effect of branching is to make the glycogen molecule more soluble and to increase the
number of non-reducing ends.
This increases the number of sites accessible to glycogen phosphorylase and glycogen synthase, both
of which act only at non-reducing ends.

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