CARBOHYDRATE METABOLISM

Dr waseem kausar.
 Introduction to Metabolism
 
 Complex substances are broken
down for energy, required metabolites,
structural components, etc.

 Cells must synthesize new complex

substances.
 
 Thousands of such reactions are
occurring simultaneously in a
single cell.
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Definitions:

Catabolism = the breakdown of

complex substances.
 
Anabolism = the synthesis of
complex substances from simpler
ones.
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General Pathways of Metabolism
-- Catabolism --
1- Breakdown of macromolecules to
building blocks -- generally hydrolytic

protein polysaccharide lipid nucleic acids

amino glucose, glycerol ribose,

acids other sugars fatty acids bases,
phosphate
-- no useable energy yield here-
only building blocks obtained
2- Breakdown of monomers to
common intermediates
amino glucose, glycerol,
acids other sugars fatty acids

pyruvate
NH4+ acetyl CoA
citric acid cycleETS/Ox PhosATP
  CO2
Oxidative processes-- produce ATP &
NADH for energy
3-Breakdown of intermediates to CO2
and electrons is accomplished
through a central oxidative pathway:

 the Citric Acid Cycle or TCA or the

Krebs Cycle.
Cycle

 This cycle leads to the production

of ATP by processes called electron
transport and oxidative
phosphorylation.
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**
proteins polysaccharides lipids
amino glucose, glycerol
acids other sugars fatty acids

NH4+ pyruvate
 
acetyl CoA Intermediates
 
citric acid cycle
  CO2
--Anabolism--
Anabolism
-- Some General Principles --
 Processes of metabolism are highly
controlled:
 Anabolism and catabolism are not
necessarily balanced - one or the
other may predominate in certain
cells or at different times depending
on cell needs
 The pathway to synthesize a
complex substance is not simply the
reverse of the degradative pathway.
 Modes of Control

1- Level of energy-
energy if low, anabolism
is unlikely or impossible
2- Level of substrates
3- Level of enzyme cofactors-
cofactors
lipoic acid, thiamine, NAD+, etc.
4- pH - affects ionization states, i.e.,
a molecule may be reactive only if in
(un)protonated state
5- Enzymes-
a) quantity- repression or induction
of expression of information in DNA
b) activity- may have inactive or less
active states, allosteric enzymes have
+ or - effectors, feedback control-
build-up of product inhibits enzyme
6- Compartmentalization - Some
enzymes and substrates restricted
to certain organelles so as to make
the substrate and enzyme available
together in right place.
7- Hormone control-
control Certain cells
are targeted by hormones, which
indirectly regulate cellular
pathways.

Definition: Hormones are small

regulatory molecules synthesized
elsewhere and delivered to target
cells.
Carbohydrate Metabolism Overview
glycogen

pentose GLUCOSE other

sugars
pyruvate

lactate acetyl CoA EtOH

TCA cycle ATP

Glycolysis
GLYCOLYSIS Glucose
ATP
hexokinase ADP
Glucose 6-phosphate
phosphogluco-
isomerase
Fructose 6-phosphate
ATP
phosphofructokinase ADP
Fructose 1,6-bisphosphate
aldolase

triose phosphate isomerase

Dihydroxyacetone Glyceraldehyde
phosphate 3-phosphate
 Glyceraldehyde 3-phosphate
glyceraldehyde NAD+ + Pi
3-phosphate NADH + H+
dehydrogenase
1,3-Bisphosphoglycerate
  ADP
phosphoglycerate kinase ATP
3-Phosphoglycerate
phosphoglyceromutase
2-Phosphoglycerate
enolase H2O
Phosphoenolpyruvate
ADP
pyruvate kinase ATP
Pyruvate
 Glycolysis
What is glycolysis?
 Ten step metabolic pathway to
convert glucose into two molecules
of pyruvate and two molecules
each of NADH and ATP.
 All carbohydrates to be catabolized
must enter the glycolytic pathway.
- Glycolysis is central in generating
both energy and metabolic
intermediaries.
 IMPORTANCE OF GLYCOLYSIS
Site = cytoplasm of ALL THE CELLS
means = sweet splitting
only source of energy of RBCs
Glucose entry into the cells by
1= Na+ independent facilitated
2= Na+ dependent
provide carbon skeleton for non essential amino acids and
glycerol part of fat.
Reversible reaction for gluconeogenesis
CERTAIN DISEASES
hemolytic disease = PK deficiency
Fatigue syndrome = PFK deficiency
-Pyruvate can be further processed:

a) anaerobically to lactate in muscle

and in certain micro-organisms or

b) anaerobically to ethanol
(fermentation) or

c) aerobically to CO2 and H2O via the

citric acid cycle.
-- Alcohol fermentation and
anaerobic glycolysis (lactate) are
not
efficient energy providers since this
glycolytic pathway yields little ATP.
-- Aerobic processing of pyruvate
through the citric acid cycle
and respiration gives a more
complete oxidation and a much
higher ATP yield.
- Glycolysis provides pyruvate,
a precursor for respiration, the
Glycolysis has two stages.

A. An energy investment phase.

Reactions, 1-5. Glucose to two
glyceraldehyde 3-phosphate
molecules. Two ATPs are invested.
B. An energy payoff phase.
Reactions 6-10. two glyceraldehyde
3-phosphate molecules to two
pyruvate plus four ATP molecules.
-- A net of two ATP molecules overall
plus two NADH.
Phase I. Energy Investment.
 
1- Glucose is phosphorylated. Glucose
enters a cell through a specific
glucose transport process. It is
quickly phosphorylated at the
expense of an ATP. The investment of
an ATP here is called “priming.”
Enzymes = hexokinase or
glucokinase
 ATP ADP

glucose glucose 6-phosphate

o
∆G ' = -16.7 kJ/mole
Reaction:

 first energy investment

 highly exergonic, ∆ G°´= -16.7
kJ/mole, (essentially irreversible)
Hexokinase
 found in all cells of every
organism
 low specificity for
monosaccharides (simple sugars)
i.e., other monosaccharides can be
phosphorylated by hexokinase.
 relatively high affinity for glucose,
KM = 0.1 mM
 inhibited by its product, glucose
6-phosphate
Glucokinase

 found in liver
 high KM (~10mM) for glucose
 not inhibited by glucose-6-
phosphate
 most effective when glucose level
in blood is high, i.e., right after meal.
2- Isomerization of glucose 6-
phosphate
Enzyme = phosphoglucoisomerase

glucose 6-phosphate fructose 6-phophate

aldose to ketose isomerization

reversible, ∆ G°′ = 1.7 kJ/mole
3- Second phosphorylation
Enzyme = phosphofructokinase

ATP ADP

fructose 1,6
bisphosphate
-second ATP investment
-highly exergonic, essentially
irreversible, ∆ G°´= -14.2 kJ/mole
- highly regulated,
regulated modulating carbon
flux through glycolysis in response
to energy and carbon requirements
4- Cleavage to two triose
phosphates
Enzyme = aldolase
HC=O H2COP
HCOH O=C
HCOP + CH2OH
H
glyceraldehyde
dihydroxyacetone
3-phosphate phosphate
where P = phosphate
 cleaves a 6C sugar to 2 3C sugars
 ∆ G°´= +23.8 kJ/mole, driven by
-- mechanism: keto at C2 of
F-1,6-bis P condenses with ε-amine
of lys at active site of enzyme
(Schiff base intermediate), aiding in
carbon-carbon bond cleavage.
H H
HC-OP H -H2O HC-OP
C=O + HN C=N
HCOH HCOH
ENZ ENZ

bond to be broken bond easily

broken now
5- Isomerization of dihydroxyacetone
phosphate
Enzyme = triose-phosphate isomerase
H2C-OH

C=O

CH2-O- P
dihydroxyacetone glyceraldehyde
phosphate 3-phosphate
 allows interconversion of two triose
phosphate products of aldolase cleavage
only glyceraldehyde phosphate can
be used further in glycolysis.
 aldose-ketose isomerization similar
to phosphoglucoisomerase rxn
allows dihydroxyacetone phosphate
to be metabolized as
glyceraldehyde 3-phosphate
reversible, ∆ G°´= +7.5 kJ/mole.
This is important in gluconeogenesis
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End of First Phase:
- Production of two glyceraldehyde
3-phosphate molecules from one
glucose molecule with the
expenditure of two ATPs.
- Therefore: the energy yields of the
following steps are multipled by two.
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Second Phase:
6- Oxidation of glyceraldehyde
3-phosphate
Enzyme= glyceraldehyde-3-phosphate
dehydrogenase O
OPO
O NAD NADH O
OPOH C=O
+ O HCOH
H2C
O- P
glyceraldehyde 3-phosphate 1,3 bisphosphoglycerate
-addition of phosphate, oxidation,
production of NADH, formation of
high energy compound
- First high energy compound
generated = beginning of payoff.
- product is an acylphosphate, a
fused
carboxylic-phosphoric acid
anhydrate, which has a very high
free energy of hydrolysis.
- reversible rxn, ∆ G°´ = +6.3 kJ/mole
because this fused group retains
some of the energy produced by the
oxidation of the aldehyde to the
carboxylic acid.
-- reaction produces important
reducing compound NADH
= nicotinamide adenine dinucleotide,
reduced form. H

[Core NAD+ is
recycled and not
used up in
metabolism.]
Mechanism of G3P Dehydrogenase
7- Transfer of phosphate to make ATP
Enzyme = phosphoglycerate
kinase
O=C-O- P O=C-OH P
HC-OH + P HC-OH + P
H2C-O-P P H2C-O-P P
Adenosine Adenosine
1,3PG ADP 3-phosphoglycerate ATP
- first substrate level phosphorylation,
yielding ATP
- 2 1,3 bis PG yield 2 ATPs, thus so far
ATP yield = ATP input
- high free energy yield, ∆ G°´=
-18.8kJ/mole drives several of the previous
8- Phosphate shift setup
Enzyme= phosphoglycerate mutase

- shifts phosphate from position 3 to 2

- reversible, ΔG°′ = + 4.6 kJ/mole
- mechanism involves phosphorylated
enzyme intermediate with the
formation of 2,3 bisphosphoglycerate
 9- Generation of second very high
energy compound by a dehydration
Enzyme = enolase

-- little energy change in this

reaction,
ΔG°′ = +1.7 kJ/mole because the
- the energy is locked into the high
energy unfavorable enol
configuration by phosphoric acid ester

- upon later hydrolysis of phosphate:

H high energy low energy

O O
-C=C-  -C-C-

This energy is recovered the next step.

10- Final generation of ATP
Enzyme = pyruvate kinase
P
O H ADP ATP O
-
OOC-C=CH -
OOC-C-CH3
 phosphoenolpyruvate pyruvate
 - second substrate level
phosphorylation yielding ATP
- highly exergonic reaction,
irreversible, ΔG°′ = -31.4 kJ/mole.
- rxn is so exergonic because the enol
in PEP is transformed to a keto in
pyruvate.
- drives several previous reactions.
- pyruvate is the primary product
of glycolysis
- pyruvate kinase is a highly
regulated enzyme.
Bookkeeping:
Bookkeeping
- 2 ATPs from each glyceraldehyde
3-phosphate = total of 4 per original
glucose in second phase.

- 2 molecules of NADH also produced.

- 2 ATPs were invested in the first

phase of glycolysis.

Glycolysis: Invest 2 ATP 4 ATP 

net 2 ATP and 2 NADH
Summary of Energy Relationships
for Glycolysis 
Input = 2 ATP
 1. glucose + ATP  glucose-6-P
 2. fructose-6-P + ATP  fructose 1,6
bisphosphate
Output = 4 ATP + 2 NADH
1. 2 glyceraldehyde 3-P + 2 Pi + 2 NAD+
2 (1,3 bisphosphoglycerate) + 2 NADH
2. 2 (1,3 bisphosphoglycerate) + 2 ADP
2 (3-P-glycerate) + 2 ATP
3. 2 PEP + 2 ADP  2 pyruvate + 2 ATP
Net = 2 ATP and 2 NADH
 Energy Yield From Glycolysis

glucose 6 CO2 = -2840 kJ/mole

 
2 ATPs produced = 2 x 30.5 =
61 kJ/mole glucose
 
Energy yield = 61/2840 = 2%
recovered as ATP
- subsequent oxidation of pyruvate and
NADH can recover more of the free
energy from glucose.
Fate of Product of Glycolysis- Pyruvate
- Pyruvate is at a central branch point
in metabolism.

Recall: 
Aerobic pathway - through
citric acid cycle and respiration;
this pathway yields far more energy
and will be discussed later.
NADH + O2  NAD+ + energy
Pyruvate + O2  3CO2 + energy
Two anerobic pathways:

 - to lactate via lactate dehydrogenase

- to ethanol via ethanol dehydrogenase

- Note: both use up NADH produced

so only 2 ATP per glucose consumed
1. Lactate Fermentation
Enzyme = Lactate Dehydrogenase
COO- COO-
C=O + NADH + H+  H-C-OH + NAD+
CH3 CH3
pyruvate lactat
- Note: uses up all the NADH
(reducing equivalents) produced in
glycolysis.
 Helps drive glycolysis by using up
NADH
 Reversible so pyruvate can be
regenerated in alternative
metabolism
 Lactate fermentation important in
red blood cells, parts of the retina,
and in skeletal muscle cells during
strenuous exercise.
 Important in plants and in
microbes growing in absence of O2.
-- Lactate Dehydrogenase (LDH) has
multiple forms. It is an isozyme.
Two polypeptides M and H come
together to form LDH. It is a tetramer
so a mixture is formed:
 
M4, M3H, M2H2, MH3 and H4

M M M H H H H H H H
M M M M MM M H
H H
 Skeletal muscle and liver contain
predominantly the “M” forms;
heart the “H” forms. During and
after myocardial
infarction (heart
attack), heart
cells die releasing
LDH into the
circulation.

 Diagnostic.
Summary Glucose
of Reactions 2 ATP
2 NADH
2 pyruvate
2 NADH 2 NADH
anaerobic anaerobic
2 ethanol + CO2 2 lactate

2 acetyl CoA + 2 CO2

  O2 aerobic
4 CO2 + 4 H2O
-- REGULATION OF GLYCOLYSIS --

Three irreversible kinase reactions

primarily drive glycolysis forward.
 
 hexokinase or glucokinase
 phosphofructokinase
 pyruvate kinase

These enzymes regulate glycolysis

as well.
 
1. HEXOKINASE and GLUCOKINASE
– Discussed
HEXOKINASE
 Phosphorylation of glucose.

Inhibited by its product, glucose

6-phosphate, as a response to
slowing of glycolysis
GLUCOKINASE

 liver enzyme with high KM for

glucose so most effective when
glucose levels are very high
 not inhibited by glucose 6-phosphate
sensitive to high glucose in
circulation from recent meal
 it decreases high level of glucose
in blood by taking glucose into liver
2. PHOSPHOFRUCTOKINASE
 rate limiting for glycolysis
 an allosteric multimeric regulatory
enzyme.
 Measures adequacy of energy levels.

 Inhibitors: ATP and citrate

high energy
 Activators: ADP, AMP, low energy
and fructose 2,6 bisphosphate
 
 ATP inhibits phosphofructose
activity by decreasing fructose
6-phosphate binding
 AMP and ADP reverse ATP inhibition

 Fructose 2,6 bisphosphate is a very

important regulator, controlling the
relative flux of carbon through
glycolysis versus gluconeogenesis.
- It also couples these pathways to
hormonal regulation.
3. PYRUVATE KINASE PEP + ADP
pyruvate + ATP
 An allosteric tetramer

- inhibitor: ATP
- inhibitors: acetyl CoA and
fatty acids (alternative fuels for
TCA cycle)

- activator: fructose 1,6-

bisphosphate (“feed-forward”)
 Phosphorylation (inactive form) and
dephosphorylation (active form)
under hormone control.

Also highly regulated at the level of

gene expression
(“carbohydrate loading”)