Carbohydrates

Metabolism of Carbohydrate

Cyberjaya University College of Medical Sciences

Prof Dr. Noor Aini AH 1
 Overview of Metabolism
protein polysaccharides lipids
ADP + Pi ADP + Pi ADP + Pi
ATP ATP ATP
amino acids hexoses
fatty acids
ADP + Pi pentoses
ADP + Pi ADP + Pi
ATP ADP + Pi
ATP
ATP
pyruvate ATP
urea
acetyl-CoA ADP + Pi
urea O2
ATP
cycle
electron transport
e- chain
citric acid oxidative
cycle phosphorylation
CO2

ATP 2
Overview of Catabolic Processes
Proteins
Proteins Carbohydrates
Carbohydrates Fats
Fats

Amino
Aminoacids
acids Simple
SimpleSugars
Sugars Fatty
Fattyacids
acids

Glycolysis
Glycolysis

Pyruvate ATP
Pyruvate

Acetyl
AcetylCoA
CoA

Citric
Citricacid
acidcycle
cycle

Oxidative
Oxidativephosphorylation
phosphorylation

ATP
3
 Release of Chemical Energy
• Cellular
respiration
occurs in three
sets of reactions:
glycolysis, the
citric acid cycle,
and the electron
transport chain
(oxidative
phosphorylation)

Session 2007/2008 4
 Glycolysis
Glycolysis
 Glucose  pyruvate/lactate
 It occurs in the cytosol
 Glycolysis – aerobic & anaerobic (does not
require oxygen to proceed)
 1st stage of glycolysis - glucose is
phosphorylated at two places, requiring ATP
 2nd stage - the 6-carbon glucose is split into two
3-carbon pyruvate molecules
 ATP is synthesized at the second stage of
glycolysis at two places

5
 Role of Glycolysis
- provide ATP
- precursor for ribose sugar
- glycerol 3-phosphate   triacylglycerol
- pyruvate   FA synthesis
amino acid synthesis

6
Reactions Involved in Glycolysis - I
glucose
ATP
hexokinase
ADP
glucose-6-P
6 carbon
Priming stage
stage
fructose-6-P
ATP phosphofructokinase
Requires
ADP Energy !!
fructose-1,6-diP

2 molecules
glyceraldehyde-3-P dihydroxyacetone-P

7
Reactions Involved in Glycolysis - II
glyceraldehyde-3-P
NAD+ Pi
Glyceraldehyde-3-P-
NADH + H+ dehydrogenase
1,3-diphosphoglycerate
ADP Phosphoglycerate kinase
ATP
3-phosphoglycerate 33 carbon
carbon
stage
stage
2-diphosphoglycerate
Double
Double this
this
H2 O
3-phosphoenolpyruvate since
since two
two
ADP Pyruvate kinase
pyruvate
pyruvate
ATP are
are made.
made.
pyruvate
8
 Overall Glycolysis

glucose + 2 ATP + 2 ADP + 2 Pi + 2 NAD+

10 enzymes

2 pyruvate + 2 NADH + 2 H2O + 4 ATP

Net energy produced is 2 ATP

In addition, the two pyruvate ions can go on to
the citric acid cycle to produce more energy.
9
 Fate of Pyruvate
Glycolysis pathway is similar in all organisms.

What happens to pyruvate will vary

significantly.
In our cells, under aerobic conditions, pyruvate
is converted to acetyl CoA in the mitochondria.

Under anaerobic condition, pyruvate is

converted to lactate
pyruvate Acetyl CoA
O dehydrogenase O
|| complex ||
CH3-C-COO- + CoA-SH CH3-C-S-CoA + CO2

NAD+ NADH 10
Regulation of Glycolysis

_ Glu 6-P

Phosphofructokinase 2
F2,6-BP, AMP
+
_
ATP, Citrate

ATP,
_ asetil KoA
Alanine
+ F1,6-BP Glucagon
Session 2007/2008 11
 Regulation of the glycolytic pathway
 The regulatory enzymes of the pathway are hexokinase,
phosphofructokinase and pyruvate kinase.

Hexokinase
 Strongly inhibited by the product reaction, glucose 6-P.
 Concentration of glucose 6-P high, the enzyme activity reduced.
 Glucokinase, liver enzyme of hexokinase not inhibited by glucose 6-
p. Concentration increased when glucose level in liver increased.
Insulin also regulates the activity.

Phosphofructokinase
 The rate-limiting enzyme. The reaction of phosphofructokinase
commits the cell to the metabolism of glucose by glycolysis. (Check
the pathway)

Pyruvate Kinase
 Inhibited by ATP concentration. Stimulated by insulin & high glucose.
12
 Aerobic Respiration

• When oxygen
is available,
pyruvate
moves from
cytosol into
mitochondria.

Session 2007/2008 13
 Anaerobic
Anaerobic Glycolysis
Glycolysis
 In the absence of O2, lactate formed (eg. RBC,
muscle of a sprinter  muscle cramps)
 Pyruvate ---> Lactate (NADH -->NAD+; LDH)
 Source of NADH (G3-P --> 1,3-BPG)
 Formation of NAD+ is important for glycolysis to
go on
 Formation of ATP 100x faster than aerobic
respiration (citric acid cycle and oxidative
phosphorylation)
 Lactate will return to the liver to be converted to
pyruvate and to undergo gluconeogenesis - Cori
Cycle

14
Cori Cycle

15
 Overall
Overall reaction
reaction for
for Glycolysis
Glycolysis
Glucose + 2ATP + 2 ADP + 2PO4- + 2NAD+

2 Pyruvate + 2 NADH + 2H2O + 4 ATP

Net Energy : 2ATP
Anaerobic : no NADH

16
KREBS CYCLE

17
Krebs Cycle (overview)

18
 Function of Krebs cycle:

 Release of chemical energy (NADH,

FADH2, GTP/ATP)
 Its intermediates are precursors of many
important compounds (Succinyl CoA =
heme synthesis, OAA  aspartate, an
amino acid)

19
 Krebs cycle
 Acetyl CoA combines with oxaloacetate to form
citrate.
 A series of reactions regenerate oxaloacetate
and produce ATP, NADH + H+, FADH2, and
carbon dioxide.
 This cycle can be repeated as long as oxygen
and pyruvate are available

20
                                                                         

Session 2007/2008 21
 Energy & the Citric Acid Cycle
acetyl CoA citrate H2 O

oxaloacetate
cis-aconitate H2 O
NADH

malate isocitrate
CO2
H2O +
NADH
fumarate
-ketoglutarate

FADH2 Coenzyme A
succinate succincyl
CoA
GTP CO2 + NADH
+ Coenzyme A GDP
22
 Pyruvate dehydrogenase

 Pyruvate  Acetyl CoA

 Multi enzyme complex
 Pyruvate dehydrogenase/pyruvate
decarboxylase
 Dihydrolipoyl transacetylase
 Dihydrolipoyl dehydrogenase

23
Regulation of TCA Cycle

24
 Citrate synthase

 Acetyl CoA + OAA  citrate

 An allosteric enzyme
 Positive effector: ADP
 Negative effectors: ATP, NADH,
succinyl CoA, acyl CoA derivative , fatty
acids

25
 Isocitrate dehydrogenase:
rate limiting TCA

 CO2
 NADH (need Mg2+)
 ADP , positive effector
 ATP and NADH negative effector

26
 -Ketoglutarate dehydrogenase
 Multi enzyme complex:
-ketoglutarate dehydrogenase/decarboxylase,
transsuccinylase,
lipoamide dehydrogenase
 -Ketoglutarate  Succinyl CoA
 Release of CO2
 Formation of NADH
 Coenzymes: NAD+, TPP, lipoic acid and CoA
 Negative effectors: ATP , GTP, NADH and
Succinyl CoA
 Similar structure to PDH

27
 Succinate dehydrogenase

 Succinate  Fumarate
 Formation of FADH
 Malonate is an analogue of succinate, a
competitive inhibitor of the enzyme

28
 OVERALL REACTION OF CITRIC
ACID CYCLE
 Acetyl CoA + 3NAD+ + FAD + GDP + Pi + 2H2O

2CO2 + 3NADH + FADH2 + GTP + 3H+ + CoA
Regulatory enzymes (mainly allosteric): citrate
synthase, isocitrate dehydrogenase, -
ketoglutarate dehydrogenase
Regulatory factors:
ADP/ATP and NAD+/NADH = regulate respiration
for energy formation

29
Efflux of intermediates from TCA
cycle

Session 2007/2008 30
Electron Transport Chain

31
 ETC

 Electrons are transferred through a series of

enzyme complexes on the folds of the inner
mitochondrial membrane
 Electron energy is transferred to ATP
 Oxygen receives the electrons to form water

32
 ETC complexes
 Complex I : NADH dehydrogenase/NADHQ
reductase (FMN, protein ferum-sulphur)
 Complex II: Succinate dehydrogenase/succinate
CoQ oxidoreductase (FAD, protein ferum-sulphur)
 Complex III: Cytochrome b-c1
complex/Ubiquinone-cytochrome c oxidoreductase
(cytochrome b and c1, protein ferum sulphur)
 Complex IV: Cytochrome oxidase/Cytochrome aa 3
(cytochrome a and a3, cupric ion)

33
Session 2007/2008 34
 ELECTRON FLOW

 NADH oxidized by FMN at complex I and

pass to CoQ (ubiquinone)
 FADH2 oxidized by ferum-sulphur at
complex II and pass to CoQ
 CoQ oxidized by cytochrome c at complex
III
 cytochrome c oxidized by O2 at complex IV

35
Session 2007/2008 36
 Sources of electrons
 Complex I: NADH from MDH, IDH, PDH and
-KDH and from oxidation of fatty acids,
glycolysis via malate/aspartate shuttle

 Complex II: FADH2 from SDH, and glycolysis

via glycerol phosphate shuttle, and oxidation
of fatty acids

37
 Chemiosmotic theory of oxidative
phosphorylation
 Electron flows through ETC, H+ is pumped
outside, from matrix into inter membrane space
(cytosolic side of inner membrane).
 Protons are pumped outside at complexes I, III
and IV.
 This creates a proton motive force due to pH
gradient or membrane potential
 H+ is pumped into the matrix via FO-F1-ATP
synthase, coupling with ATP synthesis and
acceptance of electrons by oxygen  oxidative
phosphorylation
38
Session 2007/2008 39
 Synthesis of ATP…….

 When electron flows through the complexes

to its final destination (O2), ATP and heat
generated

40
 NET OVERALL REACTION
 Pyruvate + 4NAD+ + FAD + GDP + Pi + 2H2O 
2CO2 + 4NADH + FADH2 + GTP + 3H+ + CoA
 Oxidation of 1 mole of glucose yields:
 4x2 NADH , 1x2 FADH2, 1x2 GTP, 2 NADH
(glycolysis), 2 ATP (glycolysis)
 24 ATP + 4 ATP + 2 ATP + 6/4 ATP +2 ATP =
38/36 ATP

41
 REGULATION OF OXIDATIVE
PHOSPHORYLATION

 If ADP increases  Oxidative

Phosphorylation increases
 Electron source (substrate)
 NADH/NAD+ and ATP/ADP
 Oxygen

42
 INHIBITORS OF ETC
Carboxin

ATP Barbiturates,
Rotenone,
FAD
Piericidin A Complex II
REDUCED e- e- e
NAD+ FMN CoQ
SUBSTRATE -
Complex I
e-
Antimycin A,
H2S, cyanide (CN-), CO, sodium azide
Dimercaprol

O2 e- Cyto a + a3 Cyto c Cyto b

e- e-
Complex IV Complex III
ATP ATP
Inhibitor of Fo-F1 synthase = oligomycin (antibiotic)
43
 Uncoupler of oxidative
phosphorylation
Lipophilic substances which are permeable
to membrane
 2,4-Dinitrophenol (slimming drug)
 Dicumarol (anticoagulant)
 Bilirubin (normal amount is not significant)

44
 Brown fat
 Tissue containing brown fat rich in
mitochondria (cytochrome)
 In cold condition, noradrenaline is
stimulated, lipase activated, fats being
oxidized ---> FADH2, NADH
 Thermogenin activated, proton enters
mitochondrial matrix, ETC activated,
FADH2, NADH oxidized, heat generated
(but with no ATP formation)
45
 BROWN FAT

Cold
Noradrenaline

Heat

Thermogenin H
+

O2 H2 O
TG

46
Glycogen Metabolism

47
 Glycogen
 Glycogen, also known as “animal starch,” is a
glucose polymer that serves as the body’s quick
energy reserve.

 The average person usually maintains enough for

one day’s needs.

O
O
O
 (1-6) glicosidic linkage).
O

O
O c
O
O
 (1-4) glicosidic linkage O

48
 Glycogen

Stored
Stored in
in the
the liver
liver and
and skeletal
skeletal muscles
muscles

49
 Glycogenesis
 Glycogenesis produces glycogen from
glucose.
 Can occur in all cells, but is an especially
important function of liver and muscle cells.
 When blood sugar level increases,
glycogenesis occur.
 Involves several enzymes converting the
glucose to glycogen (rebuilt the  (1-4) & 
(1-6) glicosidic linkage).
 Glucose-6-P converted to glucose-1-P &
finally glycogen formed.
 Blood glucose return to normal level.
50
 Glycogenolysis
 Glycogenolysis is the breakdown of glycogen to
glucose.
 Occurs in liver (and kidney and intestine) but in
muscle tissue, glucose 6-phosphate enter the
glycolysis to produced ATP – lack of glucose 6
phosphatase
 When blood sugar level reduced, glycogenolysis
induced.
 Hydrolysis of glycogen to glucose unit in liver
(hydrolysis of  (1-4) &  (1-6) glicosidic linkage).

51
GLYCOGEN METABOLISM
GLUCOSE
INSULIN
ATP
Hexokinase GLYCOGENESIS

ADP Glycogen synthase

Glu 6-P Glu I-P UDP-Glu Glycogen

Glycogen phosphorylase
GLYCOGENOLYSIS

GLUCAGON,
ADRENALINE

52
 GLYCOGEN METABOLISM

• liver glycogen maintains blood glucose when

blood glucose is low, however glycogen ~ 12
hours
•In muscle, glycogen ~ 400g (2% of muscle’s
weight
•In liver, glycogen ~100g (6% of liver’s weight)
•Need 160 g of glucose/day
•Brains uses 120 g
•Glycogen reserve (fr liver) ~ 12-24 hours.

53
54
 GLYCOGENESIS

 In liver, GLUT-2, takes in glucose

when it is high in the blood ie, after
meal
 Glucokinase converts glucose to glu 6-
P; if glu 6-P is in excess than is
needed for glycolysis, it will be
converted to glu 1-P for glycogenesis.

55
REGULATION OF GLUCOSE IN FED AND FASTING
STATE

56
Low Blood Glucose:
(Panic reaction)
Adrenaline mediates the fight or flight response

Effects similar to glucagon. It mobilizes glucose from glycogen.

In contrast, it also promotes glucose catabolism in muscle
tissue (for production of ATP for activity).

57
High Blood Glucose:
)
Insulin lowers blood glucose levels (hypoglycaemic hormone

Session 2007/2008 58
 Glycogen storage disease
 Type I (von Gierke’s): deficiency of glu 6-
phosphatase (liver)  hypoglycaemia  death
 Type II (Pompe’s): deficiency -1,4-glucosidase-
muscle, heart (baby: death < 2 yrs; adult: muscular
dystrophy)
 Type V (Mc Ardle ): deficiency of muscle glycogen
phoshorylase (muscle pain during exercise, cramp,
weak)
 Type VI (Hers): deficiency of liver glycogen
phosphorylase  hepatomegaly, acidosis,
retardation of growth

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Gluconeogenesis

60
 GLUCONEOGENESIS
 Synthesis of glucose from non carbohydrate sources,
occurs in liver and kidney:
Lactate
Glycerol
Amino acids
 Need energy (mainly ATP supplied from fatty acid
oxidation from adipose tissue; 6 moles of ATP)
 PDH becomes inactive (phosphorylated)
 During fasting when hepatic glycogen depletes,
gluconeogenesis is essential for maintenance of blood
glucose
 Hormonally controlled by insulin:glucagon

61
 Gluconeogenesis pathway
Lactate Pyruvate Oxaloacetate

Phosphoenolpyruvate

Fructose 1,6-biphosphate

Fructose 6-phosphate
*Gluconeogenesis
uses many
glycolytic enzymes Glucose 6-phosphate
but in the reverse
direction
Glucose 62
Session 2007/2008 63
Session 2007/2008 64
 /RBC
Session 2007/2008 65
Session 2007/2008 66
 GLU-ALA
GLU-ALA CYCLE
CYCLE

Glu Glu Glu

Gluconeogenesis

NAD+
Blood Glycolysis
6ATP
Urea Urea NADH
ATP 2 Pyruvate
2 Pyruvate
Kidney Prot
NH2
2 Alanine 2 Alanine 2 Alanine

Liver
Muscle
Both Cori and Alanine cycles occur during fasting or in between meals 67
Session 2007/2008 68
Session 2007/2008 69
 Regulation of gluconeogenesis
• Concentration of its precursors (lactate, glycerol, amino
acids)
• Regulatory enzymes :
Pyruvate carboxylase
PEP carboxykinase
Glucose 6-phosphatase
Fructose 1,6-bisphosphatase
- changes in allosteric effector – gluc  F2,6-BP
- covalent modification – inactivate pyruvate kinase- PEP
Note: glucagon, adrenaline and
glucocorticoids stimulate the synthesis of
these enzymes while insulin inhibits the synthesis
of these enzymes but enhances the synthesis of
glycolytic enzymes
70
Session 2007/2008 71
Enzymes regulation

Session 2007/2008 72
CARBOHYDRATE METABOLISM AFTER MEAL
CO2 + H2O + ATP (Na+ / ATPase at neuron Lactate
Acetyl CoA cell’s membrane)
Pyruvate
BRAIN Glu RBC
Carbohydrate Glu
Glycogen
-Amylase
Glucose
Rice Lactate LIVER
-amylase pancrease
disaccharidases (lactase, Pyruvate
maltase, sucrase)
TG
Glucose Insulin  Acetyl CoA
VLDL
Glucagon  CO2 + H2O
LDL
Small intestine +
Portal vein Fatty
ATPacids
Glycogen
+ Glycerol
Glucose Glu
Gly-3-P
Acetyl CoA MUSCLE
TG
Adipose cell CO2 + H2O + ATP
73
 METABOLISM OF CARBOHYDRATE DURING FASTING
CO2 + H2O + ATP (Na+ / ATPase nerve cell membrane) Lactate
Acetyl CoA Pyruvate
Pyruvate BRAIN Glu RBC
Glu Glycogen

Glycogenolysis
Rice Glucose
Gluconeogenesis TG
Gli 3-P FA
Insulin  Acetyl CoA
Glucagon  Pyruvate
CO2 + H2O + ATP
Lactate
Small
Portal vein Prot Glycogen
intestine
LIVER AA
TG Alanine MUSCLE
FA Pyruvate
Glycerol Lactate
Acetyl CoA
ADIPOSE
74
CO2 + H2O + ATP
TISSUE
Pentose Phosphate Pathway/
Hexose Monophosphate

75
 Pentose phosphate pathway

Also known as hexose monophosphate shunt

or 6-phosphogluconate pathway.
It functions not for producing energy directly,

but providing NADPH (mostly needed in fatty

acid synthesis) & ribose 5-P (nucleic acid
synthesis).

76
Divided to two steps:
1) Converting glucose 6-P to 6-phosphogluconate.
Glucose 6-dehydrogenase enzyme involved & NAD+
as a coenzyme needed. 6-phosphogluconate then
converted to ribulose-5-P and finally ribose 5-P for
nucleic acid synthesis.
2 molecules NADPH produced
2) Ribose 5-P (excess) converted into glycolytic
intermediates; 2 molecules fructose 6-P &
1 molecule glyceraldehyde 3-P.
77
 HMP (cytosol)

 2 phases: oxidative and non oxidative

 Oxidative (irreversible reactions; NADPH is
formed)
 Non oxidative (reversible reactions),
ribose 5-P returned to intermediates of
glycolysis
Role of Pentose Phosphate Pathway in
the generation of NADPH
 H2 O H2O2 (peroxide, free radicals)
Glutathione peroxidase
Glucose Infection
Synthesis of fatty acids GSSG GSH Drugs
& cholesterol Glutathione reductase ATP
NADPH NADP+ Hexokinase Fava beans
Other reactions
ADP
6-Phosphogluconate Glu 6-P
Oxidative
Glucose-6-P deh
NADPH + ydrogenase (G6
CO2 PD)
Ribulose 5-P
Isomerase
Non oxidative
Ribose 5-P Fru 6-P
Transketolase Transaldolase
Nucleic acids (TPP)
Glyceraldehyde 3-P
Hexose
Monophosphate Shunt
Pyruvate (glycolysis)
 Glutathione
ATP ADP ATP ADP
Cysteine Glutamyl- Glutathione
cysteine
glutamate glycine
Glutathione acts as an intracellular
antioxidant
Removes oxidants which are harmful, eg.
free radicals formation as a result of
infection, drugs, ingestion of fava beans
It maintains the integrity of cell membranes
 DEFICIENCY OF G6PD
NADP + NADPH+H+
ATP ADP
Glu 6-P 6-phospho
Glu
G6PD gluconate
 individuals with deficiency of G6PD will undergo hemolytic
anemia
RBC membrane lyses after an episode of stressed oxidants
eg. ingestion of certain drugs (eg. sulfa, primaquine and other
antimalarial), ingestion of food such as fava beans, or due to
infections
Oxidants (ROS) imposes stress on cell membrane  (GSSG)
 Heinz bodies (aggregates of metHb)  lysis jaundice
Incidence high in Asia, Africa, Mediterranean
Fructose and Galactose
Metabolism

85
 Sources of fructose

 Fruits (sucrose) , honey (fructose),

chocolate (sucrose)
 Glucose sorbitol fructose (polyol
pathway)

86
 Metabolism of Fructose
 In liver (small amount in kidney and small intestine)
 Pyruvate, lactate, glycogen, and glucose can be
generated from fructose
 Genetic disease:
 Essential Fructosuria (deficiency in fructokinase) ;
not a serious condition, fructose found in urine
 Fructose intolerance (reduced activity of aldolase B),
 hypoglycaemia (fru 1-P inhibits glycogenolysis and
gluconeogenesis), jaundice, liver failure
 Treatment : diet without sucrose and fructose

87
 Metabolism of galactose

 Galactose can be converted to glycogen, glucose via

gal 1-P
 Deficiency of galactokinase  galactosemia and
galactosuria; not serious; accumulation of galactitol
cause cataract
 Def of uridyltransferase; Classical type = accumulation
of gal 1-P dan galactitol at neuron cells, lens of eyes,
liver and kidney  mental retardation (nephropathy),
cataract, and liver impairment
 Therapy: diet without galactose or lactose

88
 Polyol pathway
Galactose Glucose (aldehyde)

NADPH + H+
Aldose reductase
NADP+
Galactitol* Sorbitol (polyol)*
Reaction is high in
NAD+ spermatozoa cells
Sorbitol dehydrogenase
NADH + H+

Fructose
*raised in eye lens (cataract) for pt with galactosemia and
diabetes mellitus;
*sorbitol high high osmotic pressure cataract and
neuronal cells dysfunction
89
 Cataract formation
Glucose (Diabetes melitus)

Glucose

Sorbitol

Fructose
H2O H2O
 high osmotic pressure and nonenzymatic
glycosylation of lens proteins
90
Thank You

91