Carbohydrate Metabolism

Mr.Tapeshwar Yadav.
II year M.Sc. Biochemistry
Mamata Medical College
Khammam
 Carbohydrates
Definition : Carbohydrates are
polyhydroxy aldehydes or ketones.
 Biological significance of
Carbohydrates
• These are major source of energy for
living organisms.
• Supplying a huge array of metabolic
intermediates for biosynthetic
reactions.
• The structural elements in cell coat
or connective tissues.
Digestion :
• Partly digested in mouth by salivary
amylase.
• In stomach there is no digestion takes
place.
• Complete digestion and absorption
will be taken place at small intestine
 Glucose transporters (GLUT)

• There are 5 types GLUT

–GLUT1: RBC
–GLUT4: Adipose tissue, Muscle
 The metabolism of glucose
• Aerobic oxidation
• Glycolysis
• Gluconeogenesis
• Pentose phosphate pathway
• Glycogenesis
• Glycogenolysis
• Uronic acid pathway
 Glycogen

Glycogenesis Glycogenolysis

Starch si s Pyruvate/
D o l y
absigesti Gly
c Lactate
orp on
tion A
Glucose oxiedrobic
Lactate, at i o
n
Amino n eo H2O+CO2
lu co is
acids, G e ne s
Glycerol -g Pentose phosphate
pathway

Ribose, NADPH
Aerobic oxidation
• C6H12O6 6H2O+6CO2 + Energy

(in the form of heat)

Glycolysis
• The anaerobic catabolic pathway by
which a molecule of glucose is
broken down into two molecules of
lactate.

Glucose → 2 Lactic acid (lack of O2)

• All of the enzymes of glycolysis

locate in cytosol.
Glycolysis overview

Glucose
Glycolysis

Pyruvate
If O2 is not available

Lactic acid
1) Glycolytic pathway :
Glucose → Pyruvate
including 10 reactions.
 (1) Glucose is phosphorylated to
Glucose 6-phosphate
HO CH2 P O CH2
ATP ADP O H
H O H 2+ H
H Mg H
OH H OH H
OH Hexokinase OH
OH OH
H OH H OH
G G-6-P

• Phosphorylated Glucose cannot get

out from cell.
• Hexokinase (having 4 Isoenzymes).
• Glucokinase, GK in liver.
• Irreversible.
 Comparison of
Hexokinase and Glucokinase
Particulars Hexokinase Glucokinase
Occurrence in all tissues only in liver
Km value 0.1mmol/L 10mmol/L
Substrate Glucose, Glucose
Fructose,
Mannose

Regulation G-6-P Insulin

(2) G-6-P is isomerised to Fructose 6-P

P O CH2 P O CH2 O CH2OH

H O H
H H OH
OH H
OH isomerase H OH
OH
H OH OH H
G-6-P F-6-P
(3) F-6-P is phosphorylated Fructose
1,6-bisphosphate
P O CH2 CH2OH P O CH2 CH2 O P
O O
PFK-1
H OH H OH
2+
Mg OH
H OH H
ATP ADP
OH H OH H
F-6-P F-1,6-BP

• The second phosphorylation

• Phosphofructokinase-1, PFK-1
 (4) F-1,6-BP cleaved to 2 Triose
phosphates
CH2 O P
C O
CH2 O P CHO
HO C H
C O + CHOH
H C OH aldolase
CH2OH CH2 O P
H C OH
CH2 O P dihydroxyacetone glyceraldehyde
phosphate, 3-phosphate,
F-1,6-BP DHAP GAP

• Reversible
(5) Triose phosphate isomerization

CH2 O P CHO
C O CHOH
phosphotriose
CH2OH CH2 O P
isomerase

DHAP GAP

G→2 molecule glyceraldehyde-3-

phosphate, consume 2 ATP .
(6) Glyceraldehyde 3-phosphate oxidised
1,3-bisphospho glycerate
Pi NAD+ NADH+H +
CHO O C O~ P
CHOH CHOH
glyceraldehyde
CH2 O P 3-phosphate CH2 O P
dehydrogenase,
glyceraldehyde GAPDH glycerate
3-phosphate 1,3-bisphosphate,
1,3-BPG
 (7) 1,3-BPG dephosphorylated to 3-
phospho glycerate
ADP ATP
O C O~ P COO-
CHOH CHOH
Phosphoglycerate
CH2 O P CH2 O P
kinase
glycerate glycerate
1,3-bisphosphate 3-phosphate

• Substrate level phosphorylation

(8) Glycerate 3-phosphate mutated
glycerate 2-phosphate

COO- COO-
CHOH CH O P
Mutase
CH2 O P CH2OH
glycerate glycerate
3-phosphate 2-phosphate
(9) Glycerate 2-phosphate →
phosphoenol pyruvate

COO- COO-
CH O P C O ~ P + H2O
CH2OH enolase
CH2
glycerate PEP
2-phosphate
(10) PEP →pyruvate

COO - ADP ATP COO-

C O~ P C O
pyruvate kinase
CH2 CH3

PEP Pyruvate

• Second substrate level

phosphorylation
• irreversible
2) Pyruvate → lactate

NADH + H+ NAD+
COO COO
C O CHOH
Lactate dehydrogenase,
CH3 CH3
LDH
Pyr Lactic acid
 Summary of Glycolysis
ADP ADP
ATP ATP
Mg2+ Mg2+
G G-6-P F- 6-P F- 1,6-BP
HK Isomerase PFK-1
Aldolase
lactate
NAD+ GAP DHAP
LDH H3PO4
NAD+
NADH+H+ glyceraldehyde
pyruvate 3-phosphate
+
ATP NADH+H dehydrogenase
pyruvate kinase glycerate
1,3-bisphosphate
ADP
ADP
PEP
Enolase ATP Phosphoglycerate
glycerate Mutase kinase
H2 O glycerate
2-phosphate 3-phosphate
Total reaction:

C6H12O6 + 2ADP + 2Pi

2CH3CHOHCOOH + 2ATP + 2H2O

Formation of ATP:
The net yield is 2 ~P or 2 molecules of
ATP per glucose.
2. Regulation of Glycolysis

• Three key enzymes catalyze

irreversible reactions : Hexokinase,
Phosphofructokinase & Pyruvate
Kinase.
1) PFK-1
The reaction catalyzed by PFK-1 is
usually the rate-limiting step of the
Glycolysis pathway.

This enzyme is regulated by covalent

modification, allosteric regulation.
bifunctional
enzyme
 2) Pyruvate kinase

• Allosteric regulation:

F-1,6-BP acts as allosteric activator ；

ATP and Ala in liver act as allosteric

inhibitors;
• Covalent modification:
phosphorylated by Glucagon
through cAMP and PKA and inhibited.
ATP ADP
Pyruvate Kinase Pyruvate Kinase- P
(active) PKA (inactive)

cAMP

Glucagon
3) Hexokinase and glucokinase

• This enzyme is regulated by covalent

modification, allosteric regulation and
isoenzyme regulation.
• Inhibited by its product G-6-P.
• Insulin induces synthesis of
glucokinase.
3. Significance of glycolysis
1) Glycolysis is the emergency energy-
yielding pathway.
2) Glycolysis is the main way to
produce ATP in some tissues, even
though the oxygen supply is
sufficient, such as red blood cells,
retina, testis, skin, medulla of kidney.
• In glycolysis, 1mol G produces 2mol
lactic acid and 2mol ATP.
§ 3 Aerobic Oxidation of
Glucose
• The process of complete
oxidation of glucose to CO2 and
water with liberation of energy as
the form of ATP is named aerobic
oxidation.
• The main pathway of G oxidation.
 1. Process of aerobic oxidation

cytosol Mitochodria

first second third

stage stage stage
G Pyr Pyr CH3CO~SCoA CO2 + H2O+ATP
glycolytic TAC
pathway
1) Oxidative decarboxylation of
Pyruvate to Acetyl CoA

COO- NAD+ NADH + H + O

C O + HSCoA H3C C ~SCoA + CO2
Pyruvate
CH3 dehydrogenase
complex
pyruvate Acetyl CoA

• irreversible;
• in mitochodria.
Pyruvate dehydrogenase complex:
E1 pyruvate dehydrogenase
Es E2 dihydrolipoyl transacetylase
E3 dihydrolipoyl dehydrogenase
thiamine pyrophosphate, TPP (VB1)

HSCoA (pantothenic acid)

cofactors lipoic Acid
NAD+ (Vpp)
FAD (VB2)
Pyruvate dehydrogenase complex:

HSCoA

NAD+
 The structure of
pyruvate dehydrogenase complex
 O- O-
N NH2 S
H3C C C HC C CH2CH2 O P O P O-
N C N C CH3 O O
C C +
H H2
TPP

H2 H2
C +2H C
H2C CH (CH2)4 COOH H2C CH (CH2)4 COOH
- 2H
S S SH SH

lipoic acid dihydrolipoic acid

 HSCoA
4'-phosphopantotheine

OH CH3 OH OH
HS CH2 CH2 NH C CH2 CH2 NH C C C CH2 O P O P O 3'AMP
O O H CH3 O O

¦Â-mercapto- ¦Â-alanine pantoic acid pyrophosphate

ethylamine
pantothenic acid
 CO2

NADH
+H+

NAD+
CoASH
2) Tricarboxylic acid cycle, TCAC
• The cycle comprises the combination of a
molecule of acetyl-CoA with oxaloacetate,
resulting in the formation of a six-carbon
tricarboxylic acid, citrate. There follows a
series of reactions in the course of which
two molecules of CO2 are released and
oxaloacetate is regenerated.
• Also called citrate cycle or Krebs cycle.
(1) Process of reactions
 CH3CO~SCoA
acetyl CoA HSCoA CH2 COO H2O CH2 COO
CO COO
HO C COO C COO
CH2 COO citrate aconitase
H2O synthase CH2 COO CH COO
oxaloacetate
NADH+H+ citrate cis-aconitate
malate dehydrogenase H2O
+
NAD aconitase
HO CH COO
malate CH2 COO
CH2 COO
Citrate cycle isocitrate CH COO
fumarase
H2O HO CH COO
+
HC COO NAD
fumarate
OOC CH isocitrate dehydrogenase
CO2
FADH2 succinate dehydrogenase NADH+H+

FAD succinyl CoA CH2 COO NADH+H+ NAD+ CH2 COO

CH2 COO syntetase
CH2 CH2
CH2 COO
CoASH GTP GDP+Pi CO~ SCoA CO2 HSCoA COCOO
succinate succinyl-CoA ¦Á-ketoglutarate ¦Á-keto-
ADP ATP
dehydrogenase glutarate
complex
 Summary of
Krebs Cycle
①

Reducing
equivalents
② The net reaction of the TCAC:

acetylCoA+3NAD++FAD+GDP+Pi+2H2O

→ 2CO2+3NADH+3H++FADH2+GTP+
HSCoA

③ Irreversible and aerobic reaction

④ The enzymes are located in the

mitochondrial matrix.
⑤ Anaplerotic reaction of
oxaloacetate

ATP ADP + Pi COOH

CH3
Biotin C H2
C O + CO2
pyruvate carboxylase C O
COOH
COOH

COOH COOH
NADPH+H+ NADP+ NAD+ NADH+H+
CH3
C H2 C H2
C O + CO2
malic enzyme CHOH malic acid DH C O
COOH
COOH COOH
(2) Bio-significance of TCAC
① Acts as the final common pathway for
the oxidation of carbohydrates, lipids,
and proteins.

② Serves as the crossroad for the

interconversion among carbohydrates,
lipids, and non-essential amino acids,
and as a source of biosynthetic
intermediates.
Krebs Cycle is at the
hinge of metabolism.
 2. ATP produced in the aerobic
oxidation
• acetyl CoA → TCAC : 3 (NADH+H+) +
FADH2 + 1GTP → 12 ATP.
• pyruvate →acetyl CoA: NADH+H+ → 3 ATP
• 1 G → 2 pyruvate : 2(NADH+H+) → 6 or
8ATP
1mol G ： 36 or 38mol ATP
（ 12 ＋ 3 ） ×2 ＋ 6 （ 8 ）＝
36 （ 38 ）
 3. The regulation of aerobic
oxidation
• The Key Enzymes of aerobic oxidation
The Key Enzymes of glycolysis
Pyruvate Dehydrogenase Complex
Citrate synthase
Isocitrate dehydrogenase (rate-limiting )
α-Ketoglutarate dehydrogenase
 (1) Pyruvate dehydrogenase complex
allosteric inhibitors: allosteric activators:
ATP, acetyl CoA, AMP, CoA,
NADH, FA NAD+,Ca2+

Pyruvate dehydrogenase
(active form)

Pi ATP

pyruvate dehydrogenase pyruvate dehydrogenase

phosphatase kinase
H2O ADP

pyruvate dehydrogenase P acetyl CoA, ADP,

Ca2+,insulin (inactive form) NADH NAD+
(2) Citrate synthase
• Allosteric activator: ADP
• Allosteric inhibitor: NADH, succinyl CoA,
citrate, ATP

(3) Isocitrate dehydrogenase

• Allosteric activator: ADP, Ca2+
• Allosteric inhibitor: ATP

(4) α-Ketoglutarate dehydrogenase

• Similar with Pyruvate dehydrogenase complex
Oxidative
phosphorylation→TCAC↑

• ATP/ADP↑ inhibit TCAC,

Oxidative phosphorylation ↓
• ATP/ADP↓ ， promote
TCAC ，
Oxidative phosphorylation ↑
4. Pasteur Effect
• Under aerobic conditions, glycolysis is
inhibited and this inhibitory effect of
oxygen on glycolysis is known as
Pasteur effect.
• The key point is NADH ：
NADH mitochondria
Pyr TCAC CO2 ＋ H2O
Pyr can’t produce to lactate.
§4 Pentose Phosphate
Pathway
1. The procedure of pentose
phosphate pathway/shunt

 In cytosol
 1) Oxidative Phase

NADPH+H+
NADP + H2O
G-6-P 6-phosphogluco- 6-Phosphogluconate
G-6-P nolactone 6-Phospho
dehydrogenase gluconolactonase NADP+

6-phosphogluconate
dehydrogenase NADPH+H+
Ribose 5-P CO2
Isomerase
Ribulose 5-P

Xylulose 5-P Epimerase

 2) Non-Oxidative Phase
Ribose 5-p Fructose 6-p Glycolysis

Fructose 6-p

Xylulose 5-p

Xylulose 5-p
Glyceraldehyde 3-p

• Transketolase: requires TPP

• Transaldolase
The net reation:
3G-6-P + 6NADP+ →
2F-6-P + GAP + 6NADPH + H+ + 3CO2

2. Regulation of pentose phosphate

pathway
 Glucose-6-phosphate Dehydrogenase is the
rate-limiting enzyme.

NADPH/NADP+↑, inhibit;
NADPH/NADP+↓, activate.
 3. Significance of pentose
Phosphate pathway
1) To supply ribose 5-phosphate for bio-
synthesis of nucleic acid;
2) To supply NADPH as H-donor in
metabolism;
 NADPH is very important “reducing
power” for the synthesis of fatty acids
and cholesterol, and amino acids, etc.
 NADPH is the coenzyme of glutathione
reductase to keep the normal level of
reduced glutathione;
H2O2 2GSH NADP+
glutathione reductase

2H2O G-S-S-G NADPH + H+

So, NADPH, glutathione and glutathione

reductase together will preserved the integrity
of RBC membrane.
Deficiency of glucose 6-phosphate
dehydrogenase results in hemolytic
anemia.
favism

 NADPH serves as the coenzyme of

mixed function oxidases (mono-
oxygenases). In liver this enzyme
participates in biotransformation.
§5 Glycogen synthesis and
catabolism
Glycogen is a polymer of glucose
residues linked by
 α (1→4) glycosidic bonds, mainly
 α (1→6) glycosidic bonds, at
branch points.
1. Glycogen synthesis (Glycogenesis)

• The process of glycogenesis

occurs in cytosol of liver and
skeletal muscle mainly.
 ATP ADP UTP PPi Gn UDP
G G-6-P G-1-P UDPG Gn+1
HK or GK UDPG glycogen
pyrophosphorylase synthase

• UDPG: G active pattern, G active donor.

• In glycogen anabolism, 1 G consumes
2~P.
• Glycogen synthase: key E.
 O

CH2OH HN

H O H
H O N
OH H O O

OH O P O P O CH2
O
H OH O− O− H H
H H
OH OH

UDPG
Branching enzyme
2. Glycogen catabolism (glycogenolysis)

Pi Gn-1 H2 O Pi
Gn G-1-P G-6-P G
Phosphorylase G-6-Pase

Phosphorylase: key E;
The end products: 85% of G-1-P and 15%
of free G;
There is no the activity of glucose 6-
phosphatase (G-6-Pase) in skeletal
muscle.
Debranching enzyme:

glucan transferase
α-1,6-glucosidase
 (α1→6) linkage
Nonreducing ends

Glycogen
phosphorylase

Transferase activity of
debranching enzyme

(α1→6) glucosidase activity of

debranching enzyme Glucose
3. Regulation of glycogenesis and
glycogenolysis
1) Allosteric regulation
In liver:
G phosphorylase
glycogenolysis
InAMP
muscle:
phosphorylase-b
Ca2+
glycogenolysis
ATP phosphorylase-a
G-6-P
2) Covalent modification

Glucagon Adenylyl
receptor G protein
epinephrine cyclase

cAMP PKA Phosphorylase

Glycogen synthase

glycogenolysis
Blood sugar
glycogenesis
 glucagon, epinephrine
active inactive
adenylate cyclase adenylate cyclase
phosphorylase b
ATP cAMP kinase
ATP Pi

inactive active
PKA PKA P
phosphorylase b H 2O
ATP ADP ADP kinase
ATP ADP
P
glycogen glycogen P
synthase synthase
phosphorylase b phosphorylase a
(active) (inactive)

H2O Pi H2O
Pi
protein
phosphatase-1

inhibitor-1 inhibitor-1
(inactive) P
ATP (active)
§6 Gluconeogenesis
• Concept:
The process of transformation of non-
carbohydrates to glucose or glycogen
is termed as gluconeogenesis.
• Materials: lactate, glycerol, pyruvate
and glucogenic amino acid.
• Site: mainly liver, kidney.
1. Gluconeogenic pathway

• The main pathway for gluconeogenesis

is essentially a reversal of glycolysis,
but there are three energy barriers
obstructing a simple reversal of
glycolysis.
1) The shunt of carboxylation of Pyr
CO2 -
GDP COO
CH O ~ P
PEP carboxykinase CH2 PEP
GTP £¨ 1/3Mt.
. 2/3cytosal£© ADP
Pyr kinase
- ATP
COO
ADP+Pi ATP CO2 COO-
C O
Biotin
C O
CH2
Pyr carboxylase
CH3
COOH £¨ Mt.£©
oxaloacetic acid pyruvate
2) F-1, 6-BP →F-6-P

ATP ADP

PFK-1
F-6-P F-1,6-BP
Fructose-
bisphosphatase

Pi H2O
3) G-6-P →G
ATP ADP

HK
G G-6-P
Glucose-6-
phosphatase

Pi H2O

• 2 lactic acid G consume

ATP?
 glucose glycogen
gluconeogenesis
G-6-P G-1-P
CYTOSOL MITOCHONDRIA
F-6-P

F-1,6BP malic acid malic acid

NAD+ glutamate glutamate NAD+

NADH+H+ ¦Á-ketoglutarate ¦Á-ketoglutarate

glyceral- NADH+H+
DHAP dehyde 3-P OAA Asp Asp OAA

GTP GTP
glycerol ADP
1.3-bisphospho-
glycerate 2/3 CO2 CO2
ADP 1/3
GDP GDP
ATP
glycerate 3-P phosphoenol phosphoenol ATP
pyruvate pyruvate
ADP CO2
glycerate 2-P PK
NAD+ NADH+H+ ATP
lactate pyruvate pyruvate
2. Regulation of gluconeogenesis
• Substrate cycle:
The interconversion of two substrates
catalyzed by different enzymes for
singly direction reactions is called
“substrate cycle”.
• The substrate cycle produces net
hydrolysis of ATP or GTP.------futile
cycle
Key enzymes of gluconeogenesis

PEP carboxykinase
Pyr carboxylase
Fructose-bisphosphatase
Glucose-6-phosphatase
 gluconeogenesis:

F-6-P ATP
Pi
F-2,6-BP
FBPase-1 PFK-1
AMP
ADP
H 2O
F-1,6-BP

glycolysis
 glucagon F-2,6-BP
PEP ADP
insulin F-1,6-BP
glucagon
Ala in liver
OAA
ATP
Pyr

acetyl CoA
3. Significance of gluconeogenesis
(1) Replenishment of Glucose by
Gluconeogenesis and Maintaining
Normal Blood Sugar Level.
(2) Replenishment of Liver Glycogen.
(3) Regulation of Acid-base Balance.
First stages
(cytosol)

Second stages
(Mt.)
Third stages
(Mt.)
 Lactic acid (Cori) cycle
• Lactate, formed by the oxidation of
glucose in skeletal muscle and by
blood, is transported to the liver where
it re-forms glucose, which again
becomes available via the circulation
for oxidation in the tissues. This
process is known as the lactic acid
cycle or Cori cycle.
• prevent acidosis ； reused lactate
 Lactic acid cycle
glucose glucose glucose

gluconeo-
genesis glycolytic
pathway
pyruvate
pyruvate
NAD+ NADH+H+

NADH+H+ NAD+
lactate lactate lactate

liver blood muscle

§6 Blood Sugar and Its
Regulation
 1. The source and fate of blood sugar

origin (income) fate (outcome)

dietary supply CO2 + H2O + energy

liver glycogen glycogen

blood sugar
3.89¡« 6.11mmol/L
non-carbohydrate
(gluconeogenesis) other saccharides

non-carbohydrates
other saccharides (lipids and some
>8.89¡« 10.00mmol/L amino acids)
(threshold of kidney)

urine glucose
Blood sugar level must be maintained
within a limited range to ensure the
supply of glucose to brain.

The blood glucose concentration is 3.89

～ 6.11mmol/L normally.
2. Regulation of blood sugar level
1 ） insulin ： for decreasing blood sugar
levels.
2 ） glucagon ： for increasing blood sugar
levels.
3 ） glucocorticoid: for increasing blood
sugar levels.
4 ） adrenaline ： for increasing blood
sugar levels.
3. Abnormal Blood Sugar Level

• Hyperglycemia: > 7.22 ～ 7.78 mmol/L

• The renal threshold for glucose: 8.89

～ 10.00mmol/L

• Hypoglycemia: < 3.33 ～ 3.89mmol/L

Pyruvate as a junction point