Gluconeogenesis

&
HMP Shunt
Dr. Ajay Kumar
Dept of Biochemistry
 Overview
• From microorganisms to humans sugar is the principle
building blocks & source of fuel
• Tissues like – brain, rbc, kidney medulla, testes &
embryo are solely dependant on glucose
• Each day brain alone requires 120 g of glucose – almost
half of what is stored as glycogen
• Supply of glucose is not always available – b/w meals,
long fasts & after vigorous exercise
• Cells synthesize glucose from non carbohydrate
precursors – sweet + new + synthesis
• lactate, pyruvate,
a.a. glycerol –
precursors
• Site – Liver &
renal cortex
Glycolysis & Gluconeogenesis
• 7 out of 10 steps are exactly
reverse of glycolysis
• 3 reactions are irreversible
– cannot be used for
gluconeogenesis
• HK, PFK-1 & PK
• These 3 reactions are
‘bypassed’ by 3 separate
reactions
 Ist Bypass – Pyruvate to PEP

• Pyruvate is transported from cytosol

to mitochondria or can be generated
from alanine by transamination
Carboxylation of Pyruvate
• Pyruvate carboxylase converts
pyruvate to OAA
• Biotin – carrier for HCO3
• Acetyl CoA is +ve effector for
pyruvate carboxylase
• OAA is converted into malate by
MDH as no transporters for it
 OAA to PEP
• Malate is transported to the cytosol
through a specific transporter in the
mit membrane
• Malate is reoxidized to OAA in the
cytosol
• OAA is then converted into PEP by
removal of CO2 & phosphorylation
• PEP Carboxykinase – reqires Mg
and GTP is the phosphoryl donor
PEP to Fructose 1,6-bisP

• Reactions continue exactly

reverse of glycolysis
 2nd Bypass - F 1,6-bisP to F 6-P

• The glycolytic PFK-1 reaction is irreversible

• F 1,6-bisP is converted to F 6-P by a separate reaction
catalyzed by Fructose 1,6-bisphosphatase
• Mg requiring enzyme hydrolyzes Carbon 1 Phosphate
• This phosphoryl gp is not transferred to ATP, instead is
liberated as Pi
Fructose 6-P to Glucose 6-P

• Reverse of glycolytic pathway

• Phosphohexose isomerase
 3 rd Bypass G 6-P to Glucose

• Glucose 6-Phosphatase hydrolyzes phosphate gp at C6 to

yield Pi
• Requires Mg, is present on luminal side of ER in liver
and kidney cells
• Muscle and brain lack this enzyme – cannot perform
gluconeogenesis
G’neogenesis – energetically expensive

• 4 ATP, 2 GTP & 2 NADH required

• This high energy cost for gluconeogenesis accounts for
its irreversibility
• Although expensive yet essential at the time of crisis
 Other glucogenic compounds

• TCA intermediates can be utilized for gluconeogenesis

by being oxidized to OAA
• Certain amino acids can be converted to pyruvate or to
certain intermediates of TCA – glucogenic a. acids
• Alanine and glutamine are principal glucogenic amino
acids in humans can be converted to pyruvate and 
ketoglutarate respectively by deamination
All of 20 essential amino acids are glucogenic
except leucine and lysine !
•Acetyl CoA derived from fatty acid oxidation can be
converted into OAA (the glyoxylate cycle) in plants, yeasts
and many fungi
•Mammals cannot produce glucose from acetyl CoA as
PDH reaction is reversible & no alternate pathway available
 Glycolysis & gluconeogenesis –
reciprocally regulated
• Gluconeogenesis occurs primarily in the liver where its
function is to provide glucose to cells whose glycogen
stores are depleted
• 3 reactions of glycolysis are highly exergonic and
irreversible – HK, PFK-1 & PK
• The 3 bypass reactions of gluconeogenesis, if operate
simultaneously, the net result would be utilization of
ATP with no biological work but heat liberation only
 Regulation – glycolysis & gluconeogenesis

• The sum of these two reactions is hydrolysis of ATP with

production of heat
• Therefore these two processes are tightly regulated so
that only one is operational at a time
 1st Regulation - gluconeogenesis
• Pyruvate can either be
converted to Acetyl CoA -
PDH (TCA) or to OAA –
Pyruvate carboxylase
(gluconeogenesis)
• Acetyl CoA is a positive
regulator for Pyruvate
carboxylase and neg
regulator for PDH
 2nd Regulation - gluconeogenesis

PFK-1 (glycolysis) FBP-1(gluconeogenesis)

• Low energy ↑AMP & • Low energy ↑AMP
ADP
• High energy – ↑ATP &
Citrate
 3rd Regulation - gluconeogenesis

• Fructose 2,6-bisphosphate – a potent activator of PFK-1

and inhibitor of FBP-1
• F2,6BP increases with increase in insulin; decreased by
glucagon
• Low blood glu → inc glucagon → decreased F26BP
• Low F26BP → low glycolysis → inc gluconeogenesis
 Overview
• In most cells the major fate for G 6-P is glycolysis
• Some amount of G 6-P does have another fate that is
oxidation via the Pentose phosphate pathway
• PPP = HMP = Phosphogluconate pathway
• NADP is the electron acceptor in this pathway to yield
NADPH
• NADPH is essential to prevent the oxidative injury to
cells by keeping glutathione reduced
• This is particularly important in cells which are directly
exposed to large amounts of oxygen – Rbc, lens, cornea
 Overview(-contd)

• NADPH is required by tissues actively synthesizing

fatty acids – adipose tissue, mammary gland & liver
• Tissues performing cholesterol and steroid synthesis
require NADPH – liver, adrenal gland, gonads
• Pentoses are used to form RNA & DNA by rapidly
dividing cells – bone marrow, intestinal mucosa, skin
HMP – 2 Phases
• Oxidative phase yields
NADPH & ribose 5
phosphate
• Non oxidative phase
yields 5 molecules of G 6-P
from 6 molecules of ribose
5-P
Step I Oxidative phase
• G 6-P to 6-Phosphoglucono-δ-
lactone (an ester)
• Glucose 6-P dehydrogenase ;
requires Mg
• One molecule of NADPH is
generated
Step II Oxidative phase

• Lactonase
Step III Oxidative phase
• 6-Phosphogluconate
dehydrogenase oxidates and
decarboxylates
• 2nd molecule of NADPH is
generated
• D-Ribulose 5-Phosphate is a
ketopentose sugar
Step IV Oxidative phase
• Phosphopentose isomerase
• D-Ribose 5-Phosphate is
aldose sugar
• Oxidative phase ends at this
point generating NADPH
(for reductive biosynthesis)
and Ribose 5-P (for
nucleotide synthesis)
 Non oxidative phase
• Tissues that require only NADPH and not Pentose
sugars, recycle them back to G 6-P through non
oxidative phase
• First step involves epimerization of Ribulose 5-P to
Xylulose 5-P
 Transketolase
• Catalyzes the transfer of a 2 C fragment from a ketose
donor to an aldose acceptor
• First it transfers C1 and C2 of Xylulose 5-P(Keto donor)
to Ribose 5-P (Aldo acceptor) to form Sedoheptulose 7-
Phosphate
• Remaining 3 C fragment of Xylulose is Glyceraldehyde
3-P
 Transaldolase
• Transfers a 3 C fragment from Sedoheptulose, condenses
it with Glyceraldehyde 3-P to yield F 6-P & Erythrose 4-P
• Transketolase acts again transferring 2 C fragment from
xylulose to erythrose forming another molecule of F 6-P
and leaving another molecule of Glyceraldehyde 3-P
• Two molecules of Glyceraldehyde 3-P condense together
to form a molecule of F 1,6-bisP
• F 1,6-bisphosphate can be converted to F 6-P and then to
G 6-P by the enzyme Fructose Bisphosphatase-1 (the 2nd
bypass reaction of gluconeogenesis
• By the sequential action of these two enzymes, in 1 cycle
six 5 Carbon sugars are converted into five 6 carbon sugars
 G6PD deficiency
• 1st enzyme in the HMP pathway
• Deficient individuals lack the supply of NADPH
• Inability to keep glutathione in reduced form
• Oxidant insult causes hemolysis – sulfonamides,
primaquine, aspirin or ingestion of fava beans
• Tropical Africa, Middle east, southeast Asia
• India – very common in state of Punjab
 Wernicke Korsakoff syndrome
• Cause 1 - Mutation in gene coding for transketolase
(requires TPP) decreasing its affinity for TPP by one
tenth
• Cause 2 – Chronic alcohol consumption causes depletion
of Thiamine (decreased intestinal absorption & increased
consumption by alcohol dehydrogenase)
• Symptoms - Mental confusion, severe memory loss and
partial paralysis
Thank You !!