Glycolysis and

Gluconeogenesis
Anaerobic processes
in the generation of
metabolic energy.
Glycolysis
What is glycolysis?
sequence of reactions that converts one
molecule of glucose to two molecules of
pyruvate with the formation of two ATP
molecules
anaerobic
Glycolysis
Why is glucose such a commonly used
fuel?
tends to exist in ring form, very stable,
doesnt generally glycosylate proteins
formed from formaldehyde under prebiotic
conditions
Glycolysis
What are the possible fates of glucose?
Glycolysis
All the intermediates in glycolysis have either
3 or 6 carbon atoms
All of the reactions fall into one of 5
categories
phosphoryl transfer
phosphoryl shift
isomerization
dehydration
aldol cleavage
Glycolysis
Entire reaction sequence may be
divided into three stages
glucose is trapped and destabilized
six carbon molecule is split into two three
carbon molecules
ATP is generated
An overview
of glycolysis
FASE PERTAMA: LIMA TAHAP REAKSI GLIKOLISIS
Glycolysis reaksi 1

glucose converted to glucose-6-PO4

ATP is needed
catalyzed by hexokinase or glucokinase
induced fit
G01= -4.0 kcal/mole
Glycolysis Reaksi 2

phosphoglucoisomerase
aldose is converted to ketose
G01=+0.4 kcal/mole
Glycolysis Reaksi 3

rate limiting enzyme allosteric

inhibited by high ATP, citric acid, long-chain fatty acids
stimulated by ADP or AMP
G01= - 3.4 kcal/mole
Glycolysis
Glycolysis reaksi 4

six carbon molecule split into 2- 3 carbon molecules

aldose and ketose
G01=+ 5.73 kcal/mole
Glycolysis reaksi 5

At equilibrium most mixture exists as

dihydroxyacetone phosphate
G01=+ 1.83 kcal/mole
Triose Phosphate Isomerase
FASE KEDUA LIMA TAHAP REAKSI GLIKOLISIS
Glycolysis reaksi 6

redox reaction
energy from redox used to form acyl
phosphate
G01= +1.5 kcal/mole
Glycolysis reaksi 6
Consists of two coupled processes
Glycolysis reaksi 7

formation of ATP substrate level

phosphorylation
Glycolysis Stage 8,9,10

phosphoryl shift uses 2,3

bisphosphoglycerate G01= +1.1 kcal/mole
dehydration G01 = +.44 kcal/mole
phosphoryl transfer G01 = -7.5 kcal/mole
Glycolysis
Fate of Pyruvate
Alcoholic Fermentation
Which organisms carry out this process?
yeast
other microorganisms
PDC requires thiamine pyrophosphate as coenzyme
NAD+ is regenerated
Lactic Acid Fermentation
Occurs in muscle cells, microorganisms
Regenerates NAD+
NAD+ and Dehydrogenases
Various dehydrogenases have a similar binding
domain for NAD+ showing their common origin
Rossman fold
Glycolysis
How can fructose be used for energy?
Glycolysis
To use galactose it must be converted to
glucose-6-PO4
Glycolysis
Glycolysis
What causes lactose intolerance?
Glycolysis
What is galactosemia?
inability
to metabolize galactose
missing galactose 1-phosphate uridyl
transferase
liver disease
development of cataracts

CNS malfunction
Galactose is converted to glucose-6P via a four step reaction
involving UDP-glucose

Hexokinase

Fructokinase
In Summary
Control of Glycolysis
Of what value is glycolysis for cells?
provides energy in form of ATP
provides building blocks for synthetic reactions

Where are most control points found?

enzymes that catalyze irreversible reactions
hexokinase
phosphofructokinase
pyruvate kinase
Gluconeogenesis
What is gluconeogenesis?
synthesis of glucose from non-carbohydrate
precursors
Why is this an important pathway?
What are some of the major precursors?
lactate, amino acids, glycerol

Where does this process occur?

liver, kidney
Gluconeogenesis
Gluconeogenesis is the synthesis of glucose
from non-carbohydrate precursors.
Glucose stores are depleted during periods of
starvation or fasting beyond a day.
Since the brain relies on glucose (120g/d) as
a source of energy, glucose must be
synthesized from molecules other than
carbohydrates.
PYRUVATE GLUCOSE
Gluconeogenesis
PYRUVATE GLUCOSE
So any molecule that can be
converted to pyruvate is considered
glucogenic
Lactate and alanine are glucogenic.
Glycerol is also glucogenic.
Outline of pathways for glucose
synthesis from the major
gluconeogenic precursors.
Reactions of glycolysis and
gluconeogenesis
glycolysis contains several
irreversible reactions
1. phosphoenolpyruvate to
pyruvate
2. fructose 6-phosphate to
fructose 1,6-
bisphosphate
3. glucose to glucose 6-
phosphate
Gluconeogenesis
The irreversible glycolytic enzymes are:
hexokinase (G =-8 kcal mol-1)
phosphofructokinase (G = -5.3 kcal mol-1 )
pyruvate kinase (G = -4.0 kcal mol-1).

The enzymes of gluconeogenesis are:

pyruvate carboxylase (ATP)
phosphoenolpyruvate carboxykinase (GTP)
fructose 1,6-bisphosphatase
glucose 6-phosphatase
The reactions unique to gluconeogenesis

1. Pyruvate + CO2 + H2O + ATP <=> Oxaloacetate + ADP +

Pi + 2H+ (= -2.1 kJ/mol).
This reaction is catalyzed by pyruvate carboxylase. Unlike the
other reactions of glycolysis and gluconeogenesis, this
reaction occurs in the mitochondrial matrix.
2. Oxaloacetate + GTP <=> Phosphoenolpyruvate +
CO2 + GDP (=+2.9 kJ/mol)
This reaction is catalyzed by phosphoenolpyruvate
carboxykinase - PEPCK.
3. Fructose-1,6-bisphosphate + H2O <=> Fructose-6-
phosphate + Pi (=-16.3 kJ/mol)
This reaction is catalyzed by fructose 1,6 bisphosphatase.
4. Glucose-6-phosphate + H2O <=> Glucose + Pi ( = -12.1
kJ/mol)
This reaction is catalyzed by glucose-6-phosphatase.
Gluconeogenesis
What is the first reaction?

O
biotin
CH3 CCO2 - + CO 2 + A TP
pyruvate
Pyruvate carboxylase
O
CH2 CCO2 - + A DP + P i
CO 2 -
Oxaloacetate
Gluconeogenesis
Why is pyruvate carboxylase of special
interest?
structural properties
contains ATP-grasp domain at N-terminal end
contains biotin-binding domain at C-terminal
end
Pyruvate Carboxylase
Pyruvate + CO2 + ATP + H2O
oxaloacetate + ADP + Pi + 2 H+

Pyruvate Carboxylase fixes CO2. Enzymes

which fix CO2. require the cofactor BIOTIN.
Biotin is a vitamin and is always involved in
CO2 fixation.
This reaction takes place in the mitochondrial
matrix.

 Biotin

Notice there are no components of ATP

in the structure of biotin.
Gluconeogenesis
What is the role of biotin in this reaction?
prosthetic group lined to -amino group of lysine
residue
carrier of activated carbon dioxide
Gluconeogenesis
Pyruvate
carboxylase is an
allosteric enzyme
activated by acetyl
CoA
needed to form
carboxybiotin
Gluconeogenesis
Carboxylation of pyruvate
occurs in the mitocondria but
next step in reaction
sequence occurs in cytosol
Oxaloacetate cant pass
out of the mitochondria.
Oxaloacetate
decarboxylated and
phosphorylated in the
cytosol.
Phosphoenolpyruvate
Carboxykinase
Gluconeogenesis
Decarboxylation of oxaloacetate is coupled with
phosphorylation by GTP
enzyme is phosphoenolpyruvate carboxykinase

O
CH2 CCO2 - + GTP
CO 2 - OPO 3 2 -
Oxaloacetate CH2 = CCO2 - + CO 2 + GD P
Phosphoenol pyruvate
Phosphoenolpyruvate
Carboxykinase

Oxaloacetate + GTP
phosphoenolpyruvate + GDP + CO2
This reaction takes place in the cytosol
PEP is now synthesized and the sum of
the two reaction is:
Pyruvate + ATP + GTP + H2O
PEP + ADP + GDP + Pi + H+.
Reversible
reaction
Gluconeogenesis
Which other steps in glycolysis are
irreversible?
conversion of fructose 1,6-bisphosphate to
fructose 6-phosphate
conversion of glucose 6-phosphate to
glucose
Gluconeogenesis
CH2 OP O3 2 - CH2 OH
C O C O
HO H fructose-1,6-bis- HO H
H OH phosphatase H OH
H OH H OH
H2 O Pi
CH2 OP O3 2 - CH2 OP O3 2 -
Fructose-1,6-bisphosphate Fructose-6-phosphate

G = -16.7 kJ mol-1
fructose-1,6-bisphosphatase is an allosteric
enzyme, inhibited by AMP and activated by ATP
Fructose 1,6-bisphosphatase
fructose 1,6-bisphosphate + H2O
fructose 6-phosphate + Pi
Fructose 1,6-bisphosphatase
is an allosteric enzyme and regulates
gluconeogenesis.

fructose 6-phosphate is easily

converted to glucose 6-phosphate.
glucose 6-phosphate + H2O
glucose + Pi.
Liver can send glucose to blood
to maintain homeostasis.
Glucose 6-phosphate is also a
precursor to glycogen.
Gluconeogenesis
Enzyme that catalyzes last reaction not found
in all tissues
liver and kidney cortex

Lumen of the ER. It takes 5 proteins to convert glucose 6-

phosphate to glucose
 Gluconeogenesis Stoichiometry
2pyruvate + 4ATP + 2GTP + 2NADH + 6H2O
glucose + 4ADP + 2GDP +6Pi +2NAD+ + 2H+;
G = -9kcal mol-1.
Just the reverse of glycolysis, G = 20kcal mol-1.

Note: it takes 6 nucleotide triphosphate

molecules to synthesize glucose. Only 2
nucleotide triphosphate molecules are
generated from glycolysis.
So it takes four extra high phosphoryl-transfer
potential molecules to drive the unfavorable
gluconeogenesis pathway.
Regulation of Glycolysis and
Gluconeogenesis
What are some of the factors that
ensure the reciprocal regulation of these
processes?
allostericregulators of key enzymes
energy charge
fructose 2,6-bisphosphate
hormones
 Glycolysis and
gluconeogenesis are
reciprocally regulated
Insulin stimulates Glucagon stimulates

A high [AMP] indicates

that the energy charge
is low and signals the
need for ATP.

High [ATP] and [citrate]

indicate the energy
charge is high and
intermediates are
abundant.
Regulation of Glycolysis and
Gluconeogenesis

fructose 2,6-bisphosphate stimulates

PFK and inhibits fructose 1,6-
bisphosphase
controlled by insulin and glucagon and
reflects the nutritional status of the cell
Regulation of Glycolysis and
Gluconeogenesis
How do hormones influence the
enzymes associated with these
processes?
influence gene expression
change transcription rate
influence degradation of m-RNA
insulin PFK, PK
glucagon PEPCK, fructose 1,6-bisphosphatase
 The Cori Cycle

Lactate from active muscle is converted to glucose in liver.

 Carl and Gerty Cori
Nobel Prize in Physiology and medicine
1947

for their discovery of the course of the catalytic

conversion of glycogen
Lactate and alanine are glucogenic

In muscle alanine is produced from

pyruvate by transamination.
pyruvate + glutamate alanine + -ketoglutarate
In the liver alanine is converted back to
pyruvate.
In active muscle lactate builds up, passes
through the blood and is converted to
pyruvate in the liver.
Thus, part of the metabolic burden of
active muscle is shifted to the liver.