DR.

KHIN MAR AYE

HOD
BIOCHEMISTRY UNIT
FACULTY OF MEDICINE
 Overview of Carbohydrate
metabolism
Carbohydrates are composed of carbon and water
and have a composition of (CH2O)n.
Most abundant organic molecules in nature
Provide most of the energy in the diet of most of the
organisms
Storage form of energy in the body
Serve as cell membrane component (mediate some
forms of intercellular communication) & structural
components
 The major nutritional role of carbohydrates is to
provide energy and digestible carbohydrates provide
4 kilocalories per gram

Metabolism = Anabolism + Catabolism

Anabolism ---- Synthesis

Catabolism --- Breakdown
 Metabolism the sum of all the chemical
changes occurring in a cell , a tissue or the body
Catabolism- the breakdown of food stuffs to
simple molecules
Anabolism- the synthesis of biomolecules from
simple precursors
Catabolism - Breakdown
Proteins Amino Acids,
Starch Glucose

Anabolism - Synthesis
Amino Acids Proteins,
Glucose Starch
 Metabolic pathways
1. Anabolic pathways
-Synthesis of compounds

2. Catabolic pathways
-Oxidative processes, release free energy

3. Amphibolic pathways
-involved in both breakdown and buildup of molecules
-Links between anabolic and catabolic pathways
eg. KCAC
 Three main pathways for energy production
Glycolysis
Citric acid cycle
Oxidative-Phosphorylation
Important sugars
Glucose Major fuel of all tissues of mammals
- Universal fuel

Glycogen Storage form of glucose

Ribose - Nucleic acids, Nucleotides,

Nucleosides

Galactose - Lactose of milk, glycolipids and

glycoproteins
 GLUCOSE- Obligate Fuel
CNS/Brain
Dependent on glucose as
primary source of fuel
120 grams of glucose / day =
480 kilocalories

RBC
Dependent on glucose
Lack mitochondria
Basic Steps involved in Energy
production
1 Glycolysis

2 Acetyl CoA Formation

3 Krebs Cycle

4 Electron Transport System

Learning Objectives of Glycolysis
Reactions of glycolysis
Regulation of glycolysis
Inhibitors of glycolysis
The oxidation of pyruvate
 GLYCOLYSIS
Definition
set of reactions that oxidized glucose to pyruvate under
aerobic or to lactate under anaerobic condition

Site Cytosol of liver and extrahepatic tissue

Glucose + 2NAD+ + 2 ADP + 2Pi

2 Pyruvates + 2NADH + 2H+ + 2ATP + 2H2O

 Overview of Glycolysis
The Embden-Meyerhof Pathway

Essentially all cells carry out glycolysis

Ten reactions - similar in most cells - but rates differ

Two phases:
-First phase converts glucose to two Gly-3-P
-Second phase produces two pyruvates

Products are pyruvate, ATP and NADH

NADH must be recycled
Overall Reactions of Glycolysis
Glucose (C6)

Hexose 6P (C6)

Triose Phosphate (C3) Triose Phosphate (C3)

NAD+
NADH+H+

Pyruvate (C3) Lactate (C3)

 Overall Reactions
Aerobic condition
2 x ADP 2 x ATP

Glucose 2 x Pyruvate

2 x NAD+ 2 x NADH + 2H+

Anaerobic 2 x ADP 2 x ATP

condition
Glucose 2 x Lactate
 Stages of Glycolysis
Priming stage

Splitting stage

Oxidoreduction-phosphorylation stage
(Energy yielding stage)
1.Priming stage
- Glucose is phosphorylated by glucokinase in
liver and hexokinase in extrahepatic tissues.

- 2 mol: of ATP are used to form G6P and

Fructose 1,6 bisphosphate.
Priming stage
2.Splitting stage
-6 carbon F 1,6 BP splitted into two mol: of
Glyceraldehyde 3P

3.Oxidation-reduction phosphorylation stage

-Two mol: of Glyceraldehyde 3P are converted
into two molecules of pyruvate or lactate
Splitting stage
Energy
releasing
stage
 Energy yielding
Aerobic glycolysis
Conversion of glucose to 2 pyruvate 4 ATP

Respiratory chain linked oxidation of 2NADH

2 x 3 = 6 ATP
Total energy = 10 ATP

Used 2 mol: of ATP in priming stage

Net 10 2 = 8 ATP
Anaerobic glycolysis
Conversion of glucose to 2 lactate produced 4ATP

Used 2 mol: of ATP in priming stage

Net 2 ATP
 Metabolic significance
1.Principal route of glucose utilization to produce
acetyl CoA and oxidation in KCAC

2.Provide 8 ATP in aerobic glycolysis and 2 ATP

in anaerobic glycolysis.

3.Provide ATP in the absence of oxygen

allows skeletal muscle to get energy when
aerobic respiration is insufficient.
4.RBC -90% , eye tissues -84% of energy is
derived from glycolysis.
5.Conversion of 1,3 BPG to 2,3 BPG decrease in
oxygen affinity and shift the oxyhaemoglobin
dissociation curve to the right
6.Glyceraldehyde 3 P used for fat (TAG)
synthesis.
7.Modified reversible pathway is gluconeogenesis.
 Regulation of Glycolysis
Key enzymes
1.Glucokinase / Hexokinase
2.Phosphofructose kinase (PFK)
3.Pyruvate kinase

Control Allosteric control

- Covalent modification short term control
(Reversible phosphorylation)
- Induction / Repression of enzyme synthesis
{long term control}
 Allosteric control
Allosteric activators
AMP (Liver, Muscle) F2,6 BP (Liver)
+ +
PFK1
F6P F1,6 BP
_ _

ATP (Liver, Muscle) Citrate (Liver, Muscle)

Allosteric inhibitors
 Specific effectors of glycolysis

Enzyme Inhibitor Activator

Hexokinase G6P none

PFK ATP, citrate, PEP ADP, AMP,

cAMP
F6P, F2,6BP,
Pyruvate kinase ATP none
Phosphatase AMP ATP,Citrate
 Covalent modification

Active PFK2 Dephosphorylated

Inactive PFK2 Phosphorylated

Pyruvate kinase(a) Dephosphorylated

Pyruvate kinase(i) Phosphorylated

Insulin dephosphorylates the key enzymes.

Glucagon, epinephrine phosphorylates the key
enzymes through cAMP dependent protein kinase.
 Fate of Pyruvate
Oxaloacetate
pyruvate
Biotin carboxylase
ADP

CO2 ATP
LDH
Ethanol Pyruvate Lactate

NAD+ NADH +H+ NADH +H+ NAD+

NAD+
CO2
TPP PDH
NADH +H+
Acetyl CoA
 Pyruvate + NADH + H+ LDH Lactate + NAD+
All muscle lactate is transferred to the liver where it is
turned back to glucose

Pyruvate Py carboxylase Oxaloacetate

ATP, Biotin ,CO2
Pyruvate Py dehydrogenase Acetyl CoA + CO2

NAD+ TPP NADH+H+

Pyruvate Acetaldehyde Ethanol

CO2 NADH+H+ NAD+

 Pyruvate dehydrogenase (PDH) reaction
Pyruvate is first transported into mitochondria via a specific
transporter on the inner membrane and then oxidized to
acetyl-CoA by pyruvate dehydrogenase complex.
Source of acetyl CoA for TCA and fatty acid synthesis
Reaction- Oxidative decarboxylation, irreversible
Site - mitochondrial matrix

Pyruvate PDH complex Acetyl CoA +CO2

TPP,CoA,Lipoate,
NAD+ FAD NADH+H+

5 Coenzymes - TPP, Lipoic acid, CoA, FAD, NAD+

Regulation of pyruvate dehydrogenase
Irreversible reaction must be tightly controlled
1. Allosteric Inhibition
- inhibited by products: acetyl-CoA and NADH
- inhibited by high ATP
2. Allosteric activation by AMP
Ratio ATP/AMP important

Covalent modification-
Active form- Dephosphorylated by PDH phosphatase
Inactive form- Phosphorylated by PDH kinase
 Learning outcome on Glycolysis &
Fate of Pyruvate
Define metabolism
Distinguish between anabolism & catabolism
Describe the glycolytic pathway
State the energy requiring & energy producing steps
in the glycolytic pathway.
Explain the metabolic fate of pyruvate.
 Krebs Citric Acid Cycle (KCAC)
Tricarboxylic Acid (TCA)

Cellular respiration can be divided into 3 stages

Stage I All the fuel molecules are oxidized to generate a

common two-carbon unit, acetyl-CoA
Stage II The acetyl-CoA is completely oxidized into CO2,
with electrons collected by NAD+ and FAD via a the citric
acid cycle
Stage III Electrons of NADH and FADH2 are transferred
to O2 via the respiratory chain (a series of electron
carriers), producing H2O and a H+ gradient, which will
promote ATP formation.
Mitochondria is the major site for fuel oxidation
to generate ATP
Starts with
Acetyl CoA
 KCAC or TCA

8 Steps Reaction
3 Steps are Irreversible
 Tricarboxylic acid (TCA) cycle
(KCAC)
It is the final common pathway where oxidative metabolism
of CHO, fatty acids & amino acids converge & their carbon
skeletons are converted to CO2 & H2O

8 steps, 3 steps irreversible

Oxaloacetate + Acetyl CoA citrate,
Isocitrate - ketoglutarate,
- ketoglutarate Succinyl CoA

Series of reactions occur totally in the mitochondrial matrix

closely to the electron transport chain (ETC)
 Catabolism of acetyl CoA liberating reducing equivalents
which are oxidized to produce energy, trapped as ATP

Provides energy as ATP (12 ATP per cycle)

Cycle operates only under aerobic conditions bec: O2 is

required as final electron acceptor.

Plays a major role in gluconeogenesis, transamination,

deamination,lipogenesis

Succinyl CoA is used in heme synthesis

Plays in both oxidative and synthetic processes ie.

amphibolic pathway
 Eg :Amphibolic pathway
Acetyl CoA oxidative p/w KCAC Energy

Synthetic p/w

Fatty acid

Succinyl CoA oxidative p/w KCAC Energy

Synthetic p/w
Heme
 Oxidation of 1 mole of Glucose
Glycolysis 2 pyruvate +2NADH + 4 ATP
= (2 x 3ATP + 4 ATP) = 10 ATP
Net = (10-2) = 8 ATP

PDH
2 Pyruvate 2 Acetyl CoA+ 2CO2 + 2 NADH
= 2 x 3 ATP= 6 ATP
KCAC
2 Acetyl CoA complete oxidation = 6NADH +2FADH2+2ATP
=(6x3 + 2x2 + 2ATP) = 24 ATP
Complete oxidation of 1mole of glucose=(8+6+24)= 38ATP
Regulation of
metabolite
flow from PDH
enz comlex

High ATP:ADP,
NADH:NAD,
Acetyl CoA:CoA
inhibit PDH complex
 Regulation of KCAC
Three enzymes of KCAC are regulated to maintain a
steady state level of ATP for the cells
Most of the regulation substrate availability & product
inhibition
Enzyme Activation Inhibition

1.Pyruvate AMP, CoA, ATP, Acetyl CoA,

dehydrogenase NAD+,Ca2+ NADH,Fatty acids

2. Citrate synthase ADP NADH, Citrate,

ATP, Succinyl CoA

3.Isocitrate Ca2+, ADP ATP

dehydrogenase
 Learning outcome on KCAC
Outline the KCAC (TAC) pathway & state the
irreversible reactions in the pathway.
Give the reactions leading to degradation of acetyl
CoA resulting in liberation of CO2
Describe the steps which result in formation of
reducing equivalents.
Discuss the importance of the pathway in energy
production & biosynthesis
Show the yield of ATP in each turn of the cycle &
when glucose is completely degraded to CO2
 Hexose Monophosphate Shunt (HMS)
Pentose Phosphate Pathway (PPP)

Alternative pathway of
glucose oxidation

Site Cytosol of liver,

lactating mammary glands, adipose tissue,
thyroid gland, adrenal
cortex, testes and
erythrocytes
 Pathway begins with glycolytic intermediate (G6P)
3 mol: of G6P gives 3 mol: of CO2 & 3 five carbon
sugar
3 five carbon residues are rearranged to regenerate
2 mol: of G6P and one mol: of glycolytic intermediate
glyceraldehyde 3 P
Does not generate ATP
Generate NADPH and Ribose sugar
- NADPH - used in anabolic reactions requiring electrons
- Ribose 5 P - used in Nucleotide biosynthesis leading to:
DNA & RNA
 HMS pathway 2 phases
1. Oxidative phase
Reactions producing NADPH
Irreversible

2. Non-oxidative phase
Produces ribose-5-P
Reversible reactions feed to glycolysis
Oxidative phase G6PD

Non-oxidative
interconversion Transketolation
of sugars

Transaldolation

Transketolation
 NADPH producing Reactions
Glucose-6-P dehydrogenase (key enzyme) (step 1)
6-P-gluconate dehydrogenase (step 3)
 The glucose 6-phosphate DH (G6PD) reaction is
the rate limiting step and is irreversible

NADPH + H+ is formed from two separate

reactions.

Transketolase requires the coenzyme thiamine

pyrophosphate (TPP), the transaldolase does
not.
 Role of NADPH in RBC
.
2O + 2H+
2

H2O2 GSH NADP+

Glutathione Se Glutathione
peroxidase reductase

H2O GSSG NADPH +H+

H2O2 can produce reactive free radical species,

damage cell membranes, and cause hemolysis
HMS Pathway Nonoxidative phase
 Regulation of HMS pathway
Glucose-6-P dehydrogenase (Key enzyme)
First step / rate limiting step

Allosteric Regulation
- Feedback inhibited by NADPH (Strong inhibitor)

Induction of enzyme
- synthesis of G6PD is induced by insulin increased
insulin / glucagon ratio after carbohydrate meal
 G6PD Deficiency
G6PD deficiency causes haemolytic anaemia

Mutations present in some populations causes a

deficiency in G6PD with consequent impairment of
NADPH production.

Detoxification of H2O2 is inhibited, and cellular

damage results - lipid peroxidation leads to
erythrocyte membrane breakdown and hemolytic
anemia.
 Exposure to anti-malarial drugs (Primaquine)
results in increased cellular production of superoxide
and hydrogen peroxide (Primaquine sensitivity)

Other chemicals known to increase oxidant stress

Sulfonamides (antibiotic)
Aspirin and NSAIDs
Quinine
Fava beans
 Symptoms of G6PD Deficiency
Black colored urine
(Hemolysis may result in urinary excretion of Hb)
Low RBC count & low hemoglobin
(Result of high rate of hemolysis)
Elevated bilirubin
(Catabolism of heme)
 Uronic acid pathway
Conversion of glucose to glucuronic acid,
ascorbic acid (but not in all primates and
guinea pig) and pentose sugar

Alternative pathway of glucose oxidation

Does not generate ATP

Site Mainly cytosol of liver parenchymal cells

 Humans and other primates lack enzyme
gulonolactone oxidase

L-gulonolactone gulonolactone 3keto

oxidase L-gulonolactone

Ascorbic acid
Thus ascorbic acid is provided from the diet in
human being
 Importance of Uronic acid pathway
For excretion of metabolites and foreign
chemicals (xenobiotics)

UDP glucuronate is the active glucuronate for

conjugation of drugs, steroid hormones,
bilirubin.

Synthesis of proteoglycans
(mucopolysaccharides)
 Learning outcome on HMS & Uronic
acid pathway
Describe the pentose phosphate pathway (HMS)
Discuss in detail the oxidative phase of the pathway
in generation of NADPH & pentose sugars.
Outline the nonoxidative phase of the pathway & its
role in metabolism of 3C, 4C, 5C and 7C sugars.
Discuss the importance of the pathway.
Explain glucose 6P dehydrogenase (G6PD)
deficiency.
Significance of uronic acid pathway.