UNIVERSITY INSTITUTE OF

BIOTECHNOLOGY
Master of Science (Biotechnology)
Subject Name : Advanced Biochemistry
Subject Code : 23BTT603
Prepared by:
Dr. Pitambri Thakur

TOPIC: Gluconeogenesis DISCOVER . LEARN . EMPOWER

• To develop abroad insight into how different experimental techniques, contribute to our investigation of
metabolic processes in the cell.
• To appreciate how current discoveries were made and what they lead us onto next.
• To recognize the importance of advanced biochemistry for understanding, diagnosis, and treatment of
disease.
COURSE OUTCOMES

• Understudies will be able to identify molecular levels of life and cellular structures.
• Disciples will learn about different type of biomolecules that occur in nature specifically in microbes
consequently learning to understand their metabolism.
• Upon learning the various aspects of biomolecules of life students will be able to understand the building
blocks of life.
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 The synthesis of glucose from non-
carbohydrate compounds is known as
gluconeogenesis.
 The major substrates/precursors
for gluconeogenesis:
 Lactate, pyruvate, glucogenic amino acids,
propianate and glycerol.
 Site:
 Gluconeogenesis occurs mainly in the liver,
and to a lesser extent in the renal cortex.
 The pathway is partly mitochondrial &
partly cytoplasmic.
 About 1 kg glucose synthesized
everyday
 Brain & CNS, erythrocytes, testes & kidney
medulla are dependent on glucose for
continuous supply for energy.
 Human brain alone requires about 120 g of
glucose per day, out of about 160 g needed by
the entire body.
 Glucose is the only source that supplies to the
skeletal muscle, anaerobic conditions.
 During starvation gluconeogenesis
maintains the blood glucose level.
 The stored glycogen is depleted within the
first 12-18 hours of fasting.
 On prolonged starvation, the
gluconeogenesis is speeded up & protein
catabolism provides the substrates, namely
glucogenic amino acids.
 Gluconeogeenesis closely resembles
the reversed pathway of glycolysis.
 The 3 irreversible steps of glycolysis are
catalysed by the 3 enzymes.
 Hexokinase
 PFK
 Pyruvate kinase
 These three stages bypassed by alternate
enzymes specific to gluconeogenesis.
 These are:
 Pyruvate carboxylase
 Phosphoenol pyruvate carboxy kinase
 Fructose-1-6-bisphosphatase
 Glucose-6-phosphatase
 Takes place in two steps pyruvate
carboxylase is a biotin dependent
mitochondrial enzyme that converts
pyruvate to oxaloacetate in presence of ATP
& CO2
 This enzyme regulates
gluconeogenesis & requires acetyl CoA
for its activity.
 Oxaloacetate is synthesized in the
mitochondrial matrix.
 It has to be transported to the
cytosol.
 Due to membrane impermeability,
oxaloacetate cannot diffuse out of the
mitochondria.
 It is converted to malate & transported to
cytosol.
 In the cytosol, oxaloacetate is
regenerated.
 The reversible conversion of oxaloacetate to
malate is catalysed by MDH, present in
mitochondria & cytosol
 In the cytosol, phosphoenolpyruvate
carboxykinase converts oxaloacetate to
phosphoenol pyruvate.
 GTP or ITP (not ATP) is used in this
reaction and the CO2 is liberated.
 For the conversion of pyruvate to
phosphoenol pyruvate, 2ATP equivalents
are utilized.
 Phosphoenolpyruvate undergoes the reversal
of glycolysis until Fructose 1,6-bisphosphate is
produced.
 The enzyme Fructose 1,6-bisphosphatase
converts Fructose 1,6-bisphosphate to Fructose
6-phosphate & it requires Mg2+ ions.
 This is also a regulatory enzyme.
 Glucose 6-phosphatase catalyses the
conversion of glucose 6-phosphate to glucose.
 It is present in liver & kidney but absent in
muscle, brain and adipose tissue.
 Liver can replenish blood sugar through
gluconeogenesis, glucose 6- phosphatase is
present mainly in liver.
Regulation of gluconeogenesis
 The carbon skeleton of glucogenic amino
acids (all except leucine & lysine) results in
the formation of pyruvate or the
intermediates of citric acid cycle.
 Which, ultimately, result in the synthesis of
glucose.
Glucose-
Alanine
Cycle
 Glycerol is liberated in the adipose tissue
by
the hydrolysis of fats (triacylglycerols).
 The enzyme glycerokinase (found in
liver & kidney, absent in adipose tissue)
activates glycerol to glycerol 3- phosphate.
 It is converted to DHAP by glycerol 3-
phosphate dehydrogenase.
 DHAP is an intermediate in
 Oxidation of odd chain fatty acids & the
breakdown of some amino acids (methionine,
isoleucine) yields a three carbon propionyl
CoA.
 Propionyl CoA carboxylase acts on this in the
presence of ATP & biotin & converts to methyl
melonyl CoA
 Which is then converted to succinyl CoA in the
presence of B12.
 Succinyl CoA formed from propionyl CoA
enters gluconeogenesis.
 Definition:
 It is a process in which glucose is converted to
Lactate in the muscle and in the liver this lactate
is re-converted to glucose.
 In an actively contracting muscle, pyruvate is
reduced to lactic acid which may tend to
accumulate in the muscle.
 To prevent lactate accumulation, body utilizes
cori cycle.
 This lactic acid from muscle diffuses into the
blood.
 Lactate then reaches liver, where it is
oxidised to pyruvate.
 It is entered into gluconeogenesis.
 Regenerated glucose can enter into blood
and then to muscle.
 This cycle is called cori cycle.
 Gluconeogenesis & glycolysis are reciprocally
regulated
 One pathway is relatively inactive when the
other is active.
 Regulatory enzymes:
 Pyruvate Carboxylase.
 Fructose-1,6-bisphosphatase.
 ATP.
 Hormonal Regulation of Gluconeogenesis.
 It is an allosteric enzyme.
 Acetyl CoA is an activator of pyruvate
carboxylase so that generation of
oxaloacetate is favored when acetyl CoA
level is high.
 Citrate is an activator.
 Fructose-2,6-bisphosphate & AMP are
inhibitors.
 All these three effectors have an exactly
opposite effect on the phosphofructokinase
(PFK).
 ATP:
 Gluconeogenesis is enhanced by ATP.
 Glucagon & glucocorticoids increase
gluconeogenesis
 Glucocorticoids induce the synthesis of
hepatic amino transferases &
provides substrate for
gluconeogenesis.
 The high glucagon-insulin ratio favors
induction of synthesis of gluconeogenic
enzymes (PEPCK, Fructose-1,6-bisphosphatase
& glucose-6-phosphatase).
 At the same time, synthesis of glycolytic
enzymes HK, PFK & PK are depressed.
 SUMMARY
• In this Lecture, we studied the carbohydrate metabolism in
general.
• We understood the process of carbohydrates absorption.
• We also discussed the various processes of carbohydrate
metabolism such as glycolysis, kreb cycle and ETC etc.

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 REFERENCES
Books
• Alberts, B. Johnson, A. Lewis, J. Raff, M. and Walter, P. 2008. Molecular Biology of the cell, 5th Ed., John
Wilson, USA, ISBN: 10: 0815341059
• George, M. Malacinski, David Freifelder 1998. Essentials of molecular biology, 4th Ed., Jones & Bartlett
Publishers, ISBN- 10: O867201371
• Voet and Voet, 2016 Fundamentals of Biochemistry 2nd Edition, John Wiley & Sons, ISBN: 978-1-118-91840-1
Websites
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•http://www.namrata.co/glycolysis-lecture-1/ http://www.namrata.co/glycolysis-lecture-2/
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•http://www.namrata.co/regulation-of- glycolysis/
•http://www.namrata.co/glycolysis-subjective- questions-solved/

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THANK YOU
For queries
Email: [email protected]

For queries
Email: [email protected]