Carbohydrates

Gluconeogenesis

PROF.DR. Mursal O. Sheikh

PROF. OF MEDICAL BIOCHEMISTRY
Benadir University
Substrates for Gluconeogenesis
• Gluconeogenesis enables the maintenance of blood
glucose levels long after dietary glucose has been absorbed
and oxidized.
• Two cycles between tissues are recognized for
gluconeogenesis:
These cycles are only functional between the
liver and the peripheral tissues that:
do not completely oxidize glucose to CO2 and
H2O, thus releasing lactate as the end product
of glycolysis.
OR
tissues that must release alanine as the end
product of glycolysis and have O2 and
mitochondria.
The Cori Cycle
Cori Cycle:
Lactate returns to the liver from peripheral
tissues.
Alanine Cycle:
Alanine returns to the liver from the
peripheral tissues.
In the alanine cycle, NADH generated
during glycolysis cannot be used to reduce
pyruvate to lactate.
The Alanine Cycle
In peripheral tissues that participate in the “Alanine
Cycle” 6 to 8 molecules of ATP can be
formed/glucose molecule.
Because in tissues that have mitochondria, the
electrons of NADH are transported into the
mitochondria by aspartate- malate shuttle or
Glycerol phosphate shuttle, and ATP is formed by
oxidative phosphorylation.
On the other hand, in peripheral tissues that use the
Cori Cycle only 2 ATP/glucose molecules are
produced.
9
Overall Cost of the Alanine Cycle:
O2 and mitochondria are required in the peripheral tissue for
the alanine cycle.
10 ATP liver+ 6-8 (ADP + Pi) muscle+ O2--->10 (ADP + Pi) live +
6-8 ATP
muscle

(Alanine --------------> glucose) Cost = 2 - 4 ATPs

Overall Cost of the Cori Cycle:
6 ATP liver +2 (ADP + Pi) RBC -------------> 6 (ADP + Pi) liver + 2
ATP
RBC

(Lactate --------------> glucose) Cost = 4 ATPs

Therefore the alanine cycle seems energetically
more efficient than the cori cycle.
The liver needs 6 ATPs to convert either alanine or
lactate to glucose.
However, the alanine cycle presents the liver
with amino nitrogen, which must be removed
as urea.
4 ATP molecules are needed for each urea
molecule synthesized in the liver (2 NH2
groups of urea are provided by 2 molecules of
alanine).
The “liver” is the most important gluconeogenic
tissue.

The kidney has similar capacity, but the liver is four

times bigger than the kidneys, thus the liver
contributes more to the maintenance of blood glucose
levels by gluconeogenesis than the kidneys.

Certain muscle fibers may have the capacity for

limited gluconeogenesis.
Since adults have 18 times more muscle mass than
liver, glucose synthesis in muscle may eventually
become important.

However, muscle tissue lacks “glucose 6-

phosphatase”, which catalyzes the last step of the
gluconeogenesis pathway.

Thus, any gluconeogenesis occurring in muscle

provides glucose for glycolysis or helps to replenish
glycogen stores in muscle and does not provide
“free glucose” for the maintenance of blood glucose
levels (Imp.).
..
 I. Glucose Synthesis From Lactate
(Cori Cycle)
Overall:
2 lactate + 6 ATP ------------> glucose + 6 ADP + Pi

Many enzymes of the glycolytic pathway are common to

gluconeogenesis but “additional” reactions are involved.

Some reactions of glycolysis are: “irreversible” and are

replaced by other “irreversible” steps of the
“gluconeogenenic” pathway.
c) Conversion of 2 moles of PEP to 1 mole of fructose-1,6
biphosphate is just the reversal of glycolytic pathway steps
• (NADH produced by the reaction lactate to pyruvate in the
• initial step is used up in converting 1,3 biphosphoglycerate to
glyceraldehyde-3 PO4)
d) Conversion of Fructose 1,6 biphosphate to Fructose 6 PO4. This
irreversible reaction is catalyzed by “Fructose-1,6
biphosphatase”.
• Conversion of Fructose 1,6 biphosphate to Fructose 6 PO4. This
irreversible reaction is catalyzed by “Fructose-1,6
biphosphatase”.
6-phosphofructo-1-kinase
(during glycolysis)
• Fructose 6 PO4 Fructose 1,6 bi PO4

Fructose 1,6 biphosphatase

(during gluconeogenesis)
..
V) Fructose 6-PO4<-------------------> glucose 6 PO4
Phosphoglucose isomerase
This is a reversible reaction (functions both in
glycolysis and gluconeogenesis).

irreversible
(VI) Glucose 6 PO4 ---------------------------> Glucose
glucose-6-phosphatase
Glucose 6 phosphatase is a hepatic enzyme and is
used in
gluconeogenesis (instead of glucokinase in
glycolysis).
..
 II. Glucose Synthesis from Amino Acids
(The Alanine Cycle)
All amino acids except leucine and lysine can supply carbons
for the net synthesis of glucose by gluconeogenesis.

If catabolism of “amino acids” leads to pyruvate or

oxaloacetate
formation, then net glucose synthesis can occur.

Catabolism of amino acid feeds carbons into the TCA cycle at

various points.

If “net” synthesis of a TCA cycle intermediate occurs from an

amino acid catabolism, then net synthesis of oxaloacetate to
glucose will occur.
 - ketoglutarate + aspartate glutamate+ oxaloacetate

 This reaction is not anaplerotic because there is no net

synthesis of a TCA intermediate. Here  - ketoglutarate,
an intermediate of the TCA cycle is utilized in the reaction.
The relationship between Glucose Synthesis from Amino
Acids and the Urea Cycle (Fig. 15.38)
In the liver:
a)2 amino acids ----------> 2 pyruvates + 2 amino nitrogens

1 Glucose

b)In the liver this N is converted to urea.

Two alanine molecules (the most important gluconeogeneic
amino acid) are converted to 2 pyruvates in the cytoplasm
by removing NH2 groups

The NH2 group removed from 1 alanine molecule in

cytoplasm, is picked up by α-ketoglutarate which is
converted to glutamate and enters mitochondria. In the
mitochondria, glutamate converts back to α-ketoglutarate
by releasing NH4. This NH4 then becomes part of the urea
cycle (fig 15.38).

The NH2 group removed from the other alanine is similarly

transported by the α-ketoglutarate-glutamate pair to
mitochondria, and then NH2 group is transferred from
glutamate to oxaloacetate to make aspartate.
 Glucogeneic amino acids:
give rise to a net synthesis of pyruvate or
oxaloacetate.
Glucogeneic-Ketogeneic amino acids:
yield pyruvate or oxaloacetate, and also the ketone
bodies acetoacetate or acetyl Co A.
( Thereonine, isoleucine, tyrosine and tryptophan)
 Regulation of Gluconeogenesis:
Enzymes that “go around” the irreversible steps of
glycolysis are involved in the regulation of
gluconeogenesis.
Inhibition of glycolysis at its regulatory sites or repression of
synthesis of glycolytic enzymes (glucokinase and pyruvate
kinase) at their regulatory sites increases the effectiveness of
gluconeogenesis enzymes.

Turning on gluconeogenesis is accomplished mostly by

shutting off glycolysis.

Regulation of Gluconeogenesis
I.Allosteric regulation
II.Hormonal regulation
. Allosteric Regulation:

Fatty acid oxidation promotes glucose synthesisby:

I a. An increase in steady state concentration of

mitochondrial acetyl CoA.

Increased concentration of acetyl CO A is a

+ ve allosteric effector of the enzyme pyruvate
carboxylase (pyruvate--------> OAA)
II.Increased Fatty acid oxidation produces more ATP, which
leads to a decrease in AMP.

Low AMP:
(a)will not activate “6 phosphofructo-1- kinase” and
“pyruvate kinase”.
( b) Will not Inhibit “Fructose 1,6 biphosphatase”.

This will favor “gluconeogenesis”and inhibit glycolysis.

On the other hand, a shortage of O2 or fatty acid oxidation

will result in inhibition of oxidative phosphorylation.
The liver will then turn from gluconeogenesis to glyclysis
..
II. (a) Glucagon and insulin also regulate
gluconeogenesis by influencing “phosphorylation of
hepatic enzymes” which are subject to covalent
modulation.

(b) Glucagon and Insulin also “induce” or

“repress” the synthesis of hepatic enzymes of
glycolysis and gluconeogenesis.
 II. Hormonal Control of gluconeogenesis
Hormones act by:
I.Regulating the supply of fatty acids to the liver.
II.Regulating the enzymes of both the gluconeogeneic and
glycolytic pathway.

I. Glucagon increases lipolysis in adipose tissues, which

leads to:
Increased fatty acid oxidation by the liver.
Increased glucose synthesis in the liver by increased
supply of fatty acids to the liver. Insulin opposes lipolysis
and has the opposite effect.
II. (a) Glucagon’s Influence by Covalent Modulation
1) By inactivating glycolytic enzyme “pyruvate
kinase”.
(Glucagon activates adenylate cyclase------> cAMP----
> cAMP dependent protein kinase ----> inactivates
pyruvate kinase by phosphorylation)

Inactivation of “pyruvate kinase” stimulates

gluconeogenesis by blocking the “futile” conversion of
PEP ---------> Pyruvate.
2) Glucagon also stimulates gluconeogenesis by: working
through cAMP, which decreases the concentration of
fructose 2,6 bi PO4 in the liver by stimulating the
phosphorylation of the bifunctional enzyme
“6-PF-2-kinase/fructose 2,6 biphosphatase”.

Phosphorylation of 6-PF-2-kinase/fructose 2,6

biphosphatase:
Inactivates “kinase” that converts F6PO4 to F2,6 bi
PO4 and also
Activates “phosphatase” that hydrolyzes F2,6 bi PO4
to
F6PO4.
II b. Glucagon and insulin also have long-term
effects on levels of hepatic enzymes of glycolysis and
gluconeogenesis

Increased Glucagon/insulin ratio results in:

Increased Gluconeogenesis
and
Decreased Glycolysis
Increased Insulin/glucagon ratio :
Has an opposite effect.
This is accomplished by induction and repression
of synthesis of key enzymes of these pathways.

Increased Glucagon signals:

(a)the induction in the liver of “increased”
synthesis of
PEP carboxykinase and glucose 6-phosphatase,
(b) repression of synthesis of glucokinase and
pyruvate kinase.
 Alcohol Oxidation Inhibits Gluconeogenesis

Oxidation of alcohol in the liver produces a large load of

NADH that have be transported to the mito. by the Mal-Asp
shuttle.

Excess NADH in the cytoplasm interferes with gluconeogenesis

because it forces the equlibrium of the reaction catalyzed by
lactate dehydrogenase and malate dehydrogenase towards lactate
and malate respectively (Clin. Corr. 15.10).

This inhibits gluconeogenesis by limiting the availability of

pyruvate and oxaloacetate for the reaction catalyzed by pyruvate
carboxylase and PEP carboxykinase.
..
Since acetyl CoA formed by the oxidation of fatty acids
cannot be converted to pyruvate or oxaloacetate , it is
impossible to make glucose from fatty acids.

An exception applies to fatty acids with methyl branches

e.g., or fatty acids with odd # carbon atoms., catabolism of
these fatty acids yields propionyl CoA:

Propionyl CoA is a precursor for gluconeogenesis, because it

yields oxaloacetate by anaplerotic reaction.
Propionyl CoA is also produced during oxidation of valine and
isoleucine and in the conversion of cholesterol to bile acids.

Also, Phosphorylation of glycerol (obtained from

triglycerides) produces glycerol 3-phosphate, which is
converted in dihydroxy acetone phosphate, an intermediate of
gluconeogenesis. See Fig. 15.33. (Thus a minor part of fats
can be converted to glucose).

Depending on the nutritional state, the last stage of glycolysis

can compete with gluconeogenesis and convert dihydroxy
acetone phosphate into lactate or into pruvate for complete
oxidation.
..