Gluconeogenesis

• Definition of Gluconeogenesis :
Synthesis of new glucose from non carbohydrate sources these sources include:
Lactate, pyruvate, glycerol, TCA cycle intermediates and amino acids.
• Function of Gluconeogenesis:
• Gluconeogenesis supplies the body with glucose during starvation , low
carbohydrate diet or when liver glycogen is depleted (liver glycogen is adequate
for 12-18 hours). Adult human body required 160 g glucose/day
• Glucose is the only source of energy for brain nervous tissues RBCs kidney
medulla and skeletal muscles under aerobic conditions.
• Brain is especially sensitive to the decrease of glucose level (the daily
glucose requirement of the brain in a typical adult human being is
about 120 g).
• Red blood cells use only glucose as a fuel.
• Glucose is the precursor of milk sugar in mammary gland.
• Gluconeogenesis cleans the blood from the waste products of
 Gluconeogenesis
• Location: In the cytosol and mitochondrial of both liver 90%
and kidney10%.
• Reaction: The steps of the gluconeogenesis are mainly the
reversal of glycolysis except for the three irreversible kinases
as follows:
Enzyme of gluconeogenesis Enzyme of glycolysis

Glucose 6 phosphatase Glucokinase

Fructose 1,6 bisphosphatase Phosphofructokinase 1

Pyruvate carboxylase Pyruvate kinase

Phosphenol pyruvate carboxykinase.
• Seven out of ten reactions of gluconeogenesis are exact

reversals of glycolysis.

• Three steps in glycolysis (No 1,3,10 ) are irreversible and thus

must be bypassed in gluconeogenes.

Gluconeogenisis from pyruvate by reverse of glycolysis
 Pyruvate to phosphoenol pyruvate
1- This conversion is done by dicarboxylic acid shuttle and needs 2 enzymes:
• Pyruvate carboxylase : present in the mitochondria.
• Phosphoenol pyruvate carboxykinase: present in the cytosol and
small amount in the mitochondria).
2- Pyruvate should pass first from cytosol to mitochondria by special
transporter.
3- Pyruvate is then converted into oxaloacetate by pyruvate
carboxylase in mitochondria.
4.Essential cofactor of pyruvate carboxylase is biotin, which serves as a carrier of
CO2.
 Pyruvate (oxaloacetate) to phosphoenol pyruvate
4.The mitochondrial membrane is impermeable to oxaloacetate so
oxaloacetate is converted malate by malate dehydrogenase.
5- Malate is transported to cytosol, where it is converted again into
oxaloacetate by cytoplasmic malate dehydrogenase.
6- Oxaloacetate is converted into phosphoenol pyruvate by
phosphoenolpyruvate carboxykinase (PEP).
• Pyruvate never goes in the course of citric acid pathway to reach
malate, because this pathway
needs insulin which is deficient
during gluconeogenesis.
 Phosphoenolpyruvate carboxykinase reaction

• Reaction takes place in the cytosol.

• Oxaloacetate is simultaneously decarboxylated and
phosphorylated by phosphoenolpyruvate carboxykinase.
• Fructose 1,6 bisphosphate to fructose-6-
phosphate:
This reaction is catalyzed by the enzyme fructose 1,6
bisphosphatase.
• Glucose -6- phosphate to glucose:
This reaction is catalyzed by the enzyme glucose -6-
phosphatase.
glucose-6-phosphate + H2O  glucose + Pi

Glucose-6-phosphatase
6 CH OPO 2 CH2OH
2 3
5 O O
H H H H
H H2O H
4
OH H 1
OH H + Pi
OH OH OH OH
3 2
H OH H OH
glucose-6-phosphate glucose
The Net Reaction of Gluconeogenesis

2 Pyruvate + 2 NADH + 4 ATP + 2 GTP + 6 H2O  Glucose +

2 NAD+ + 4 ADP +2 GDP + 6 Pi + 2H+

Glucose 6-phosphatase is present in the liver and kidney, but absent
in brain and muscle. Thus, glucose produced by gluconeogenesis in
the liver, is delivered to the blood stream to uptake by brain and
Muscle
Gluconeogenisis from pyruvate by reverse of
glycolysis
Continue: Gluconeogenisis from pyruvate by reverse of glycolysis
Subcellular Locations of Gluconeogenic Enzymes

• Gluconeogenesis enzymes are cytosolic except:

• (1) Glucose 6-phosphatase (endoplasmic
reticulum)
• (2) Pyruvate carboxylase (mitochondria)
• (3) Phosphoenolpyruvate carboxykinase
(cytosol and/or mitochondria)
 GLUCONEOGENESIS FROM VARIOUS METABOLITES
I- When lactate is the gluconeogenic precursor(e.g. after vigorous
exercise)
Firstly , lactate is converted to pyruvate catalyzed by LDH in cytosol

Step 1: Pyruvate is transported into mitochondria on the pyruvate

transporter. Within mitochondria pyruvate is converted to
oxaloacetate catalyzed by pyruvate carboxylase.
Step 2: Intramitochondrial oxaloacetate is converted to Phosphoenol
pyruvic (PEP) catalyzed by mitochondrial form of PEP
carboxykinase.
Step 3: PEP is transported out of mitochondria and continues up the
gluconeogenic pathway.
II. Amino Acids:
• Can be metabolized to either pyruvate or certain
intermediates of the citric acid cycle.
• Alanine and glutamine are of special importance as they
are used to transport amino groups from a variety of
tissues to liver deaminated to pyruvate and α-Ketoglutaric
that take gluconeogenic pathway
III- TCA Cycle Intermediates:
• Form oxaloacetate during one turn of the cycle that in turn is
converted into phosphoenol pyruvic and reverse glycolysis. So can
get net synthesis of glucose from TCA cycle intermediates.
IV- Glycerol :
can be generated by hydrolysis of triacylglycerols(fat) to yield free
fatty acids + glycerol . It is an excellent substrate for gluco-
neogenesis. Glycerol --------glycerol-3-P------- Dihydroxyaceton-
Phosate. reverse glycolysis.
 Regulation of Gluconeogenesis

• Hormonal regulation :

• Glucocorticoids e.g. cortisol stimulate gluconeogenesis by the

following mechanisms:
• Induce stimulate the synthesis of gluconeogenesis enzymes:
pyruvate carboxylase, phosphoenolpyruvate carboxykinase,
fructose 1,6 bisphosphatase and glucose 6 phosphatase.
 Regulation of gluconeogenesis

1. Glucocorticoids stimulate protein catabolism by tissues this leads

to increase the glucogenic amino acids available for gluconeogenesis.

2. Insulin Inhibits gluconeogenesis. It acts as repressor inhibitor for

synthesis of enzymes of gluconeogenesis pyruvate carboxylase.

3. Other hormones as glucagons, epinephrine stimulates

gluconeogenesis.
 Regulation of gluconeogenesis

Acetyl CoA and ATP:

• Fatty acid oxidation which increased during fasting, stimulate
gluconogenesis by:
• 1) It produces ATP molecules which inhibit glycolysis (by
inhibiting phosphofructokinase-1) and stimulate gluconeogenesis
(by stimulating fructose 1,6 bisphosphatase).
• 2) Fatty acid oxidation also produces excess acetyl CoA which
stimulates pyruvate carboxylase (gluconeogenesis) and inhibits
pyruvate dehydrogenase (oxidation).
 ALANINE AND LACTATE CYCLES
• Lactate and alanine, produced by skeletal muscle and RBCs are shifted
from the muscle to the liver. They are the major fuels for gluconeogenesis.
• Alanine-glucose cycle under normal condition, alanine resulting from
muscle protein catabolism is shifted to liver and converted to pyruvic acid,
then take gluconeogenic pathway.
• Lactate-glucose cycle (Cori cycle) in RBCs and during muscle exercise.
Cori cycle is the conversion of glucose into lactate in peripheral tissues,
followed by conversion of lactate into glucose in liver