Describe the significance of Gluconeogenesis and Illustrate the structural pathway

 Gluconeogenesis is the production of glucose from non-sugar precursors.

 Gluconeogenesis, mainly occurs in the liver, and involves the synthesis of glucose from
compounds that are not carbohydrates.
 Gluconeogenesis is defined as the biosynthetic pathway for formation of glucose de-novo (i.e.
not glucose from glycogen a regular stored form in most animals)
 Gluconeogenesis is a metabolic pathway that is actually responsible for the generating glucose from
non-carbohydrate carbon containing substrates such as pyruvate, lactate, glycerol, and glucogenic
amino acids
 Gluconeogenesis is a ubiquitous process, observed in all of living kingdom including plants, animals,
fungi and bacteria. This process is also referred to as an endogenous glucose production (EGP)
 When a cell is growing on a hexose such as glucose, and obtaining glucose for polysaccharide
synthesis, there is no problem.
 But when the cell is growing on other carbon compounds, glucose must be synthesized. This
process is called as gluconeogenesis.
 Gluconeogenesis uses phosphoenolpyruvate, which is one of the intermediates
of glycolysis, as starting material and travels backwards through the glycolytic pathway to
form glucose.
 However, it involves several enzymatic steps that do not occur in glycolysis; thus, glucose is
not generated by a simple reversal of glycolysis alone.
 The major precursors for gluconeogenesis are lactate, amino acids (which form pyruvate or
TCA cycle intermediates), and glycerol (which forms DHAP).
 The synthesis of 1 mole of glucose from 2 moles of lactate requires energy equivalent to
about 6 moles of ATP.
 Location of Gluconeogenesis
 Liver, kidney, and intestine; not in skeletal muscle. The first reaction (catalyzed by pyruvate
carboxylase) takes place in the mitochondria, whereas the rest of the reactions occur in the
cytosol.
 Steps in Gluconeogenesis


 Pyruvate carboxylase converts pyruvate to oxaloacetate in the mitochondrion.

 Oxaloacetate is converted to malate or aspartate, which travels to the cytosol and is
reconverted to oxaloacetate.
 Phosphoenolpyruvate carboxykinase converts oxaloacetate to phosphoenolpyruvate.
 Phosphoenolpyruvate forms fructose 1,6-bisphosphate by reversal of the steps of glycolysis.
 Fructose 1,6-bisphosphatase converts fructose 1,6-bisphosphate to fructose-6-phosphate,
which is converted to glucose-6-phosphate.
 Glucose-6-phosphatase converts glucose-6-phosphate to free glucose, which is released into
the blood.
 Reactions involved in Gluconeogenesis
 Step 1-conversion of pyruvate to oxaloacetate
 The irreversible carboxylation reaction is performed by pyruvate carboxylase enzymeand during this
step one ATP molecule is consumed to synthesize C-C bond. This stepoccurs in mitochondria because
the enzyme pyruvate carboxylase is a mitochondrial protein.
 Step 2-conversion of oxaloacetate to phosphoenolpyruvate
 Oxaloacetate is synthesized in the mitochondria by pyruvate kinase and is shuttled into thecytosol
where it is converted into phosphoenolpyruvate. In order for oxaloacetate to leave the mitochondria,
it must be reduced to malate. In cytosol, malate is then reoxidized to oxaloacetate.
 Step 2 is catalyzed by phosphoenolpyruvate carboxykinase(PEPCK) is enzyme. The reaction is
comprised of decarboxylation and group transfer reaction and requires GTP molecule.
 Step 3-

 Phosphoenolpyruvate is converted to 2-phosphoglycerate by enolase

 Step-4 and 5-
 2-Phsphoglycerate is converted to 3-phosphoglycerate by enzyme phosphoglycerate mutase
which is then converted to 1,3-bisphosphoglycerate by phosphoglycerate kinase
 Step6 and 7-
 1,3-BPG is converted to glyceraldehyde-3 phosphate by glyceraldehyde phosphate
dehydrogenase which is therefore converted to fructose 1,6- bisphosphate
 Step 8--conversion of fructose-1, 6-bisphosphate to fructose 6-phosphate
 This step is catalysed by fructose-1,6-bisphosphatase enzyme which essentially performthe reverse
reaction catalysed by phosphofructokinase during glycolysis.
 This is a hydrolysis reaction and is irreversible.

 Step 9 –
 Fructose-6-phosphate is converted to glucose 6- phosphate by the same isomerase used in
glycolysis.
 Step 10- Conversion of glucose-6-phosphate to glucose
 This final step in gluconeogenesis is the generation of glucose. This does not take place in the cytosol
instead, glucose 6-phosphate is transported into the endoplasmic reticulum, where it is hydrolyzed
to glucose by glucose 6-phosphatase.
 In majority of tissues gluconeogenesis ends when glucose 6-phosphate is formed from fructose 6-
phosphate since it cannot diffuse out of cell like free glucose.
 Glucose-6-phosphate releases inorganic phosphate, which produces free glucose that enters
the blood. The enzyme involved is glucose 6-phosphatase.

PRECURSORS

 The major noncarbohydrate precursors of gluconeogenesis are-

 Lactate
 Glucogenic amino acids
 Glycerol (from triacylglyceride hydrolysis)
1. Lactate: Conversion of lactate to pyruvate
 Lactate are produced by active muscle cells when the rate of glycolysis exceeds therate of oxidative
mechanisms. This
 lactate is transported to liver and converted back to pyruvate and thento glucose. The metabolic
conversionof glucose to lactate and from lactateto glucose is known as Cori cycle named after Carl
and Geti Cori.
 In Cori Cycle, lactate accumulated in

 the muscle cells is taken up by liver.The liver performs gluconeogenesisto convert lactate back to
glucose. The conversion of lactate to pyruvate is catalysed by lactate dehydrogenase enzyme.

2. Glucogenic amino acids: Glucogenic amino acids are those amino acids whosecarbon skeleton can
be used to form glucose molecules. Glucogenic amino acids contributes to gluconeogenesis by two
ways
 Some amino acids can converted directly to pyruvate.
 Few amino acids are converted to TCA intermediates that in turn metabolises tooxaloacetate.
3. Glycerol
The hydrolysis of triacylglycerol in fat cells yield glycerol and fatty acids. Glycerol may enters the
gluconeogenic pathway at dihydroxyacetone phosphate (DHAP) intermediate. In the fasting state
glycerol released from lipolysis of adipose tissue triacylglycerol is used solely as a substrate for
gluconeogenesis in the liver and kidneys.

 Glycerol is converted to glycerol 3-phosphate by the enzyme glycerol kinase and then toDHAP by
glycerol phosphate dehydrogenase. The glycerol kinase is absent in adipose tissues hence the
formed glycerol is transported to liver and participates in gluconeogenesis
NET CHARGE
2 pyruvate + 2 NADH + 4ATP + 2GTP + 6H2O + 2 H+ → Glucose + 2 NAD+ + 4
ADP +
2 GDP + 6 Pi
REGULATION





Significance of Gluconeogenesis Pathway
 Gluconeogenesis meets the needs of the body for glucose when sufficient
carbohydrate is not available from the diet or glycogen reserves.
 Glycogen stored in adipose tissue and in skeletal muscle is converted to glucose by
glycogenolysis. However the stored glycogen may not be sufficient during heavy
exercise, diabetic conditions,or during fasting etc. so during shortage, glucose is
synthesized by gluconeogenesis process.
 A continual supply of glucose is necessary as a source of energy especially for the
nervous system and erythrocytes.
 Gluconeogenesis mechanism is used to clear the products of the metabolism of other
tissues from the blood, eg: Lactate, produced by muscle and erythrocytes and glycerol,
which is continuously produced by adipose tissue.
 Write notes on Anaplerotic reactions Regulation of TCA Cycle
A) Anaplerotic reactions
 Since the TCA cycle intermediates are used for anabolism, their concentration varies
according to the needs of the cell.
 Reactions that replenish the TCA cycle intermediates are called as anaplerotic reactions or
replenishing reactions.
 Anaplerosis here means restoring various intermediates of citric acid cycle that has been
used in various reactions mentioned above in amphibolic processes of a cell
 Here the intermediates are reformed or refed into the cyclic pathway that had been
previously extracted for various biosynthesis popularly called cataplerotic reactions
 The intermediates are precursors for biosynthetic reactions.
Examples include- alpha ketoglutarate, succinyl co-A, fumarate and oxaloacetate.
 Transamination reaction converts glutamate to alpha-ketoglutarate or vic-versa
 Glutamate precursor for synthesis of other amino acids and purine nucleotides
 Succinyl co-A precursor for porphyrins
 Oxaloacetate can be transaminated to form aspartate- a precursor for other amino acids
and pyrimidine nucleotide.
 Oxaloacetate is substrate for gluconeogenesis
 TCA Cycle is a metabolic hub with hub of metabolism, with vital importance in both energy
yielding and biosynthesis processes. Thus, it is critical for the cell to maintain intracellular
concentrations of various intermediates
 Steady state (homeostasis) will only be maintained when the anaplerotic flux is balanced by
cataplerotic flux of TCA cycle intermediates during cellular metabolism
 Oxaloacetate can be considered as a primary substrate of the TCA cycle. It is replenished
from pyruvate by the gluconeogenic enzyme pyruvate carboxylase:
Pyruvate + CO2 + ATP + H2O ------Oxaloacetate + ADP + Pi
 Pyruvate carboxylase is activated in the presence of acetyl CoA
 Pyruvate can also replenish malate
 Below mentioned are few reaction that recharge the flux of intermediates used is various
biosynthetic pathways:
I.Formation of oxaloacetate from pyruvate by simple carboxylation reaction in
presence of pyruvate carboxylase
II. Formation of oxaloacetate from malate by malate dehydrogenase [malate is
formed by malic enzyme that converts pyruvate into malate]
III. Formation of oxaloacetate from aspartate by a transamination reaction
IV. Formation of alpha keto glutarate by transamination reaction of in presence of
enzyme glutamate dehydrogenase
V. Formation of Succinyl-Co-A can be done in two ways either by oxidation of odd
chain fatty acids or by methionine or isoleucine metabolism. Here first propionyl-Co-
A is formed then methyl melonyl-Co-A and succinyl-Co-A.
 Anaplerotic reactions include PEP and pyruvate carboxylase
 1. Pyruvate carboxylase is an important anaplerotic reaction that catalyzes the 1st
step of gluconeogenesis from pyruvate and is found in mitochondria.
They need ATP, CO2, Biotin and MG2+. It is a major anaplerotic enzyme
 Converts pyruvate (3C) to Oxaloacetate (4C)
 Formation of succinyl co-A from propionyl coA ( source- synthesis of
bile acid, catabolism of isoleucine,threonine and methionine)
2. PEP carboxylase is found in yeast, bacteria and plants and not in animals.
B) Regulation of TCA Cycle
REGULATORY ENZYMES-
1.Citrate synthase.
2.Isocitrate dehydrogenase.
3.α-ketoglutarate dehydrogenase
Regulation of TCA cycle is determined mainly by two things
 Product inhibition and
 Substrate availability
o Overproduction of NADH and ATP like reduced coenzyme may result in to a waste of
huge amount of metabolic energy, if the cycle allowed to run without any check
point.
o ADP is other major substrate for the cycle. Which is then converted into ATP and a
resulting reduced amount of ADP leads into accumulation of NADH, a precursor
molecule by which a number of enzymes get inhibit.
o In the TCA cycle NADH isone of the product of all dehydrogenases with one
exception that is succinate dehydrogenase.
o NADH inhibits several enzymes of TCA cycle viz. citrate synthase, α-ketoglutarate
dehydrogenase, isocitrate dehydrogenase and pyruvate dehydrogenase.
o Pyruvate dehydrogenase also gets inhibited by Acetyl-coA and α-ketoglutarate
dehydrogenase and citrate synthase get inhibited by succinyl-CoA.
o Under in-vitro condition citrate synthase and α-ketoglutarate dehydrogenase gets
inhibited BY ATP.
o Citrate act as feedback inhibitor for phosphofructokinase, an enzyme which is
involved in glycolysis and catalyses the formation of a precursor of
pyruvate i.e.fructose 1,6- bisphosphate. It prevents a constant high rate of flow when
there is an accumulation of citrate with a decrease in substrate.
o Citrate Synthase
Citrate synthase is responsible for the rate of reaction in the first step of the cycle
when the acetyl-CoA is combined with oxaloacetic acid to form citrate. It is inhibited
by high concentrations of ATP, acetyl-CoA, and NADH which indicates an already
high level of energy supply. The molecule produced in the reaction, citrate, can also
act as an inhibitor of the reaction.
 o Because citrate synthase is inhibited by the final product of the citric acid cycle as
ATP, ADP (adenosine diphosphate) works as an allosteric activator of the enzyme as
ATP is formed from ADP. Therefore, the rate of the cycle is reduced when the cell has
a high level of ATP.
o Inhibitors: NADH, ATP, succinyl-CoA, citrate
o Stimulators: ADP
Isocitrate dehydrogenase
o The enzyme isocitrate dehydrogenase is an important catalyst in the third step of the
reaction. It regulates the speed at which the citrate isomer isocitrate loses a carbon to
form the five-carbon molecule α-ketoglutarate. The coenzyme NADH is a product of
the reaction and, at high levels, acts as an inhibitor by directly displacing the
NAD+ molecules it is formed from.
o Inhibitors: NADH and ATP -
o Stimulators: NAD+ , ADP and Ca +2
o Alpha ketoglutarate dehydrogenase
o The enzyme α-ketoglutarate dehydrogenase is another important catalyst in the
fourth step of the cycle where α-ketoglutarate also loses a carbon and combines with
Coenzyme A to form succinyl CoA. The two products of the reaction, succinyl CoA
and NADH, both work as inhibitors at large concentrations.
o Inhibitors: NADH, ATP and succinyl-CoA -
o Stimulators: NAD+, ADP, AMP

Describe in detail the Glycolysis pathway and add a note on the energy yield of
Oxidation of one molecule of Glucose
 Glycolysis is the process of enzymatic break down of one molecule of glucose(6 carbon) into two
pyruvate molecules(3 carbon) with the concomitant net production of two molecules of ATP.
 Glycolysis is also known as Embden-Meyerhof pathway, given by Embden, Meyerhof and parnas
 Glycolysis is an almost universal central pathway of glucose catabolism.
 Glycolysis is anaerobic process. During glycolysis some of the free energy is released and
conserved in the form of ATP and NADH.
 Glycolytic breakdown of glucose is the sole source of metabolic energy in somemammalian
tissues and cells (RBCs, Brain, Renal medulla and Sperm cell)


 Glycolysis occurs in TEN steps.
 The process of glycolysis are divided into two phases as shown in figure 4.
 Preparatory phase (phase 1)
 Payoff phase (phase 2)

1. Preparatory phase:
In preparatory phase of glycolysis, two molecule of ATP
are invested and hexose chain is cleaved into two triose
phosphates. The energy is invested in the process of
phosphorylation of glucose. The first five reactions
constitute the preparatory phase.
 Step V: Conversion of dihydroxyacetone phosphate to glyceraldehyde 3-
phosphate. Only glyceraldehyde-3-phosphate can be directly degraded into the
subsequent steps of glycolysis. The other product, dihydroxyacetone phosphate, is
rapidly and reversibly converted to glyceraldehyde-3-phosphate by the enzyme triose
phosphate isomerase.

 After the triose phosphate isomerase reaction, the two halves of the glucose have both
yielded glyceraldehyde 3-phosphate.
 This reaction completes the preparatory phase of glycolysis. The hexose molecule has
been phosphorylated at C1 and C6 and then cleaved to form two molecules of
glyceraldehyde 3-phosphate.

 2.Payoff phase:
 In payoff phase (phase 2) of glycolysis, some of the chemical energy of glucose is
conserved in the form of ATP and NADH.
 In payoff phase the conversion of two molecules of glyceraldehyde 3-phosphate to
two molecules of pyruvate is accompanied by the formation of four molecule of ATP
from ADP.
 The remaining five reactions constitutespayoff phase.
Thermodynamics of Glycolysis:
In glycolysis, one molecule of glucose is break down into two molecules of
 pyruvatereleasing 2 ATP and 2 NADH. The overall equation of aerobic glycolysis is
Glucose + 2NAD+ + 2ATP +2Pi → 2pyruvate + 2ATP + 2NADH + 2H2O + 2H+

Illustrate the steps of Kreb’s cycle and write about the significance of this pathway
 The tricarboxylic acid cycle (TCA cycle), also known as the citric acid
cycle or the Krebs cycle, is a major energy-producing pathway in living bodies.
 The Krebs cycle, also known as the citric acid cycle or TCA cycle is a series of
reactions that take place in the mitochondria resulting in oxidation of acetyl CoA to
release carbon dioxide and hydrogen atoms that later lead to the formation of water.
 The cycle also serves in the synthesis of fatty acids, amino acids, and glucose.
 This cycle is termed the citric acid cycle as the first metabolic intermediate
formed in the cycle is citric acid.
 This cycle is also termed tricarboxylic acid (TCA) because citric acid was the first
product of the cycle.
  The citric acid cycle is the final common pathway for the oxidation of all
biomolecules.Molecules from other cycles and pathways enter this cycle through
Acetyl CoA.
 The citric acid cycle is a cyclic sequence of reactions formed of 8 enzyme-
mediated reactions.
 Location of Krebs Cycle
 The citric acid cycle occurs in the mitochondrial matrix in the eukaryotes.
 In the prokaryotes, the reaction cycle occurs in plasma membrane.
 Krebs cycle Equation/ Reaction
 The overall reaction/ equation of the citric acid cycle is:
 Acetyl CoA + 3 NAD+ + 1 FAD + 1 ADP + 1 Pi → 2 CO2 + 3 NADH + 3 H+ + 1
FADH2 + 1 ATP
 Steps-
 In eukaryotic cells, all the enzymes are present in the mitochondrial matrix
except for succinate dehydrogenase and aconitase, which are present in the inner
mitochondrial membrane.

1. Formation of acetyl-coA:
 Before entering into Krebs cycle, carbohydrates, fats and proteins are catabolized
by separate pathway to form Acetyl-coA.
  It is an irreversible oxidative decarboxylation reaction in which a molecule of
carbon in the form of CO2 is removed from pyruvate.
 In this reaction a molecule of pyruvate generate 1 NADH.
 This step is link between glycolysis and Krebs cycle.
2. Reactions of citric acid cycle
i. Formation of citrate (citric acid):

 It is a condensation reaction. Acetyl-coA condensed with oxaloacetate to form

citrate and the reaction is catalyzed by the enzyme Citrate synthase.
 Oxaloacetate play catalytic role in citric acid cycle and at the end of process
oxaloacetate is regenerated.
ii. Isomerization of citrate to Isocitrate:

 The enzyme aconitase catalyzes the isomerization of citrate to isocitrate with

intermediate cis-aconitate.
 This is a reversible reaction.
 This reaction occurs in two steps- first dehydration and second hydration.
iii. Formation of α-ketoglutarate:

 This is an oxidative decarboxylation reaction.

 The enzyme isocitrate dehydrogenase catalyzes the oxidation of isocitrate to form
α-ketoglutarate and CO2.
 In this step 1 molecule of NADH is generated. It is an irreversible reaction.
iv. Formation of succinyl-coA:
 This is also an oxidative decarboxylation reaction catalyzed by α-ketoglutarate
dehydrogenase enzyme in which α-ketoglutarate is oxidized into succinylcoA and
CO2.
 In this reaction, coA serves as carriers of succinyl group and NAD+ serves as
electron acceptor.
 One molecule of NADH is generated in this step

Formation of Succinate:
 Conversion of succinyl-coA to succinate is catalyzed by succinyl-coA synthetase
or succinic thiokinase enzyme.
 This is a substrate level phosphorylation reaction in which CoA group ultimately
donates its phosphate group to GDP forming energy rich GTP.
 A molecule of CO2 is released in this step.

vi. Formation of fumarate:

 Succinate dehydrogenase catalyzed the oxidation of succinate to from fumarate.
 It is reversible reaction.
 In this step a molecule of FADH2 is generated.

vii. Formation of malate:

 This reaction is catalyzed by fumarase (fumarate hydratase) in which fumarate is
converted into malate.
 This is a hydration and reversible reaction.
viii. Formation of oxaloacetate: regeneration of oxaloacetate:
 Malate is oxidized into oxaloacetate generating a molecule of NADH.
 This reaction is catalyzed by malate dehydrogenase enzyme.
 Oxaloacetate is regenerated in this step and combines with acetylcoA and
continues the cycle.

REGULATORY ENZYMES-
1.Citrate synthase.
2.Isocitrate dehydrogenase.
3.α-ketoglutarate dehydrogenase
Regulation of TCA cycle is determined mainly by two things
 Product inhibition and
 Substrate availability
o Citrate Synthase
Citrate synthase is responsible for the rate of reaction in the first step of the cycle
when the acetyl-CoA is combined with oxaloacetic acid to form citrate. It is
inhibited by high concentrations of ATP, acetyl-CoA, and NADH which indicates
an already high level of energy supply. The molecule produced in the reaction,
citrate, can also act as an inhibitor of the reaction.
o Inhibitors: NADH, ATP, succinyl-CoA, citrate
o Stimulators: ADP
Isocitrate dehydrogenase
o The enzyme isocitrate dehydrogenase is an important catalyst in the third step of
the reaction. It regulates the speed at which the citrate isomer isocitrate loses a
carbon to form the five-carbon molecule α-ketoglutarate. The coenzyme NADH is
a product of the reaction and, at high levels, acts as an inhibitor by directly
displacing the NAD+ molecules it is formed from.
o Inhibitors: NADH and ATP -
o Stimulators: NAD+ , ADP and Ca +2
 o Alpha ketoglutarate dehydrogenase
o The enzyme α-ketoglutarate dehydrogenase is another important catalyst in the
fourth step of the cycle where α-ketoglutarate also loses a carbon and combines
with Coenzyme A to form succinyl CoA. The two products of the reaction,
succinyl CoA and NADH, both work as inhibitors at large concentrations.
o Inhibitors: NADH, ATP and succinyl-CoA -
o Stimulators: NAD+, ADP, AMP
Krebs cycle Products
Since this is a cyclic process, the oxaloacetate formed at the end as it condenses with
acetyl CoA in the next cycle.
At each turn of the cycle,
 3 NADH,
 1 FADH2,
 1 GTP (or ATP),
 2 CO2
Significance of TCA cycle: Role of TCA cycle
i. Role in Central metabolic pathway:
 TCA cycle is a final common metabolic pathway of
carbohydrates, fattyacids and aminoacids.
 At first all these biomolecules are catabolized by their separate metabolic
pathways to generate acetyl-coA then acetyl-coA enters TCA cycle for further
metabolism in aerobic condition.
 TCA is more efficient in energy conservation than other pathways of metabolism.
ii. TCA is an amphibolic pathway:
 It plays role in both catabolism and anabolism.
Catabolic role:
 TCA is a catabolic pathway because it oxidizes acetyl-coA completely into CO2
and H2O and releases large amount of energy.
Anabolic role:
 TCA is an anabolic pathway because it provides precursors for biosynthesis of
other molecules in cells. Such as citrate, α-ketoglutarate, succinylcoA and
oxaloacetate act as precursors for biosynthesis of various molecules.
iii. Citric acid cycle is an aerobic process:
 NAD+ and FAD are electron acceptors in the TCA cycle. These are regenerated
by Electron transport chain which requires oxygen as final electron acceptor. Hence
overall TCA and ETC are aerobic process.
Functions
1. Oxidation of acetyl CoA to CO2.
2. Formation of NADH and FADH2 for entrance into the electron transport chain
and subsequent ATP generation.
3. Synthesis of several important molecules, including succinyl CoA (precursor
molecule of heme), oxaloacetate (early intermediate molecule in gluconeogenesis
and substrate for amino acid synthesis), α-ketoglutarate (substrate for amino acid
synthesis), and citrate (substrate for fatty acid synthesis).
4. It is responsible for the major share of energy release and supply during aerobic
respiration.
What is the alternate pathway for glucose oxidation where does it occur? Describe the
steps of the pathway
 The pentose phosphate pathway is a metabolic pathway parallel
to glycolysis which generates NADPH and pentoses (5-carbon sugars) as well as
ribose 5-phosphate.
 The pentose phosphate pathway is also called as the phosphogluconate pathway
or hexose monophosphate shunt.
 While it involves oxidation of glucose, its primary role is anabolic rather than
catabolic.
 It is an important pathway that generates precursors for nucleotide synthesis and
is especially important in red blood cells (erythrocytes).
Location
 Cytoplasm of cells of the liver, adrenal cortex, and lactating mammary glands. In
plants, most steps take place in plastids.
The Pathway
 Substrate: Glucose-6-phosphate. There are two distinct phases in the pathway.
The first is the oxidative phase, in which NADPH is generated, and the second is the
non-oxidative synthesis of 5-carbon sugars.


Reactions
1. The Oxidative phase
Step 1:
 Glucose-6-phosphate is oxidized to form lactone. NADPH is produced as a
byproduct of this reaction as NADP^++start superscript, plus, end superscript is
reduced as glucose-6- phosphate is oxidized. Following the oxidation of glucose-6-
phosphate, another reaction, catalyzed by a different enzyme, uses water to form 6-
phosphogluconate, the linear product.
 Enzyme: glucose-6-phosphate dehydrogenase
 Step 2:
 6-Phosphogluconolactone is hydrolyzed to 6-phosphogluconate.
o Enzyme: Gluconolactonase
o Step-3:
 6-Phosphogluconate undergoes an oxidation, followed by a decarboxylation.
CO2 is released, and a second NADPH H is generated from NADP . The remaining
+ + +

carbons form ribulose-5-phosphate.

o Enzyme: 6-phosphogluconate dehydrogenase
2. The non-oxidative phase:
o The non-oxidative phase is really handy because these reactions are reversible. This
allows different molecules to enter the pentose phosphate pathway in different
areas of the non oxidative phase and be transformed up until the first molecule of
the non-oxidative phase (ribulose-5-phosphate). Ribulose-5-phosphate is the
precursor to the sugar that makes up DNA and RNA, and is also a product of the
oxidative stage.

Step 4:
 Ribulose-5-phosphate can be converted into two different 5-carbon molecules. One is
called, ribose-5-phosphate . Ribulose-5-phosphate isn’t being divided because the
carbon count is the same in the next step.
 Ribulose-5-phosphate is isomerized to ribose-5-phosphate or epimerized to
xylulose-5-phosphate through Ribulose-5-phosphate isomerase or epimerase
respectively.
Step 5:
 Ribose-5-phosphate and xylulose-5-phosphate undergo reactions, catalyzed by
transketolase and transaldolase, forming sedoheptulose-7--phosphate and
glyceraldehyde-3-phosphate.
 Transketolase, which requires thiamine pyrophosphate, transfers two-carbon
units.
 Transaldolase transfers three-carbon units.
Step 6:
 sedoheptulose-7--phosphate and glyceraldehyde-3-phosphate then undergoes
transaldolase forming xylulose-5-phosphate and erythrose-4-phosphate
 All together forming fructose-6-phosphate and glyceraldehyde-3-phosphate
Result of Pentose Phosphate Pathway
 Oxidative portion: Irreversible.
Generates two NADPH, which can then be used in fatty acid synthesis and cholesterol
synthesis and for maintaining reduced glutathione inside RBCs.
 Nonoxidative portion: Reversible.
Generates intermediate molecules (ribose-5-phosphate; glyceraldehyde-3-phosphate;
fructose-6- phosphate) for nucleotide synthesis and glycolysis.
Regulation of Pentose Phosphate Pathway
 Key enzyme is glucose-6-phosphate dehydrogenase.
 Levels of glucose-6-phosphate dehydrogenase are increased in the liver and
adipose tissue when large amounts of carbohydrates are consumed.

Purpose of Pentose Phosphate Pathway

 Pentose phosphate pathway functions as an alternative route for glucose
oxidation that does not directly consume or produce ATP.
 The pentose phosphate pathway produces NADPH for fatty acid synthesis.
Under these conditions, the fructose-6-phosphate and glyceraldehyde-3-phosphate
generated in the pathway reenter glycolysis.
 NADPH is also used to reduce glutathione (γ-glutamylcysteinylglycine).
 Glutathione helps to prevent oxidative damage to cells by reducing hydrogen
peroxide (H O ).
2 2

 Glutathione is also used to transport amino acids across the membranes of

certain cells by the γ-glutamyl cycle.
 Generation of ribose-5-phosphate

Describe the following

Glycogenesis
Glycogenolysis

GLYCOGENESIS-
 Glycogen is the major storage form of carbohydrate in animals similar to starch
in plants.
 It is a homopolymer made up of repeated units of α- D glucose and each
molecule is linked to another by 1→4 glycosidic bond which is a link connecting the
1st C atom of the active glucose residue to the 6th C atom of the approaching glucose
molecule.
 Once there is a chain consisting of 8 to 10 glycosidic residues in the glycogen
fragment, branching begins by 1→6 linkages.
 Glycogenesis is the process of glycogen synthesis, in which glucose molecules
are added to chains of glycogen for storage.
 

Location
Glycogenesis takes place in the cytoplasm of cells in muscle, liver, and adipose tissue.
 Substrate: UDP-glucose.
 Result: Changes glucose to glycogen
Steps Involved in Glycogenesis

Glycogen synthesis requires 3 main

enzymes. Glycogenesis occurs by a
different pathway from glycogenolysis.

1. UDP-glucose formation by UDP-glucose pyrophosphorylase

2. Glycogen synthesis by glycogen synthase
3. Glycogen Branching
UDP-glucose formation by UDP-glucose pyrophosphorylase

1. Glucose is activated before polymerisation in to glycogen.

Glucose is phosphorylated into glucose-6-phosphate by the action of glucokinase or
hexokinase with conversion of ATP to ADP.
 2. Glucose-6-phosphate is converted into glucose-1-phosphate by the action of
phosphoglucomutase, passing through the obligatory intermediate glucose-1,6-
bisphosphate (isomerization).
3. Glucose-1-phosphate is converted into UDP-glucose by the action of the enzyme UDP-
glucose pyrophosphorylase. Pyrophosphate is formed, which is later hydrolysed by
pyrophosphatase into two phosphate molecules.
4. Now, the glucose can be transferred to the C4–OH group on one of the non reducing
ends of glycogen to form an α-1,4 glycosidic bond. This reaction is catalyzed by the
enzyme glycogen synthase, liberating UDP,which is then recycled by conversion to UTP

STRUCTURE OF UDP-
GLUCOSE

with adenosine triphosphate (ATP).

Glycogen synthesis by glycogen synthase
 Glycogen synthase transfers the glucosyl residue from UDP-glucose to the non
reducing terminal residues of glycogen. It is transferred to hydroxyl terminal of
C4 end of glycogen to form an α-1–4 glycosidic bond.
 This reaction is catalysed by glycogen synthase. Glycogen synthase is the
regulatory enzyme in synthesis of glycogen.

UDP-glucose + (glycogen) n residues UDP + (glycogen) n+1 residues

 Glycogen synthesis requires a primer. It can add glucosyl residues to the
glycogen chain if it contains more than four residues. It means that glycogen
synthase can only extend an existing chain. Priming function is carried out by
glycogenin.
 Glycogenin:
o It is a primer for glycogen synthesis. It is a protein which composed of
two identical subunits. It is a 37 kDa protein that is glycosylated on a
specific tyrosine residue. Glycogenin contains eight glucose units linked
by α- 1–4 linkages. These glucose molecules are added to the protein by
autocatalysis.

o Formation of glycogenin primer: The first glucose molecule is attached to

the hydroxyl group on Tyr-194 of glycogenin, which is catalysed by the
subunit of glycogenin. Then it is auto catalytically extends the glucose
chain by up to seven residues long. This glucose molecule is donated by
UDPG. In this form, glycogenin can act as a primer.
 FORMATION OF GLYCOGENIN PRIMER

 New glucose residue is attached to the primer by Glycogen synthase.

Glycogen Synthase catalyzes the transfer of glucose from UDP-glucose.
New glucose molecule will transfer to the C-4 hydroxyl group at the non
reducing end of the growing glycogen molecule.

Glycogen Branching

2. Once a chain of eight glucose monomers is formed, glycogen synthase binds to

the growing glycogen chain and adds UDP-glucose to the 4-hydroxyl group of
the glucosyl residue on the non-reducing end of the glycogen chain, forming
more Alpha(1→4) bonds in the process.
3. Glycogen synthase catalyzes only α- 1–4 glycosidic bonds. It results in to the
formation ofα- amylose. Branching is catalysed by separate enzyme called
Branching enzyme.
4. Branches are made by glycogen branching enzyme (also known
as amylo α(1:4)→α(1:6)transglycosylase), which transfers the end of the chain
onto an earlier part via α-1:6 glycosidic bond, forming branches, which further
grow by addition of more α-1:4 glycosidic units.
Significance
Glucose and its precursors are obtained through food. However under certain
conditions they might not a reliable and continuous source of energy.
The glycogenesis process is therefore a built in mechanism of the body which stores the
excess carbohydrates we consume, in the form of glycogen which could be broken down
to glucose when needed.

Glycogenolysis
 The biological degradation of glycogen is termed as glycogenolysis.
 Glycogenolysis, process by which glycogen, the primary carbohydrate stored in
the liver and muscle cells of animals, is broken down into glucose to provide
immediate energy or to maintain blood glucose levels during the times of need.
 Glycogenolysis is thus the breakdown of glycogen (n) to glucose-1-
phosphate and glycogen (n-1).
Location
Glygogenolysis takes place in the cytoplasm of cells in muscle, liver, and adipose tissue.
Result: Glucose-1-phosphate is released from the non-reducing ends of glycogen
chains.
Steps Involved
1. Glucose-1-phosphate formation from nonreducing end of glycogen by
Glycogen phosphorylase
2. Removal of α-1,6 branches from glycogen by Glycogen Debranching enzyme
3. Glucose-6-phosphate formation from Glucose-1-phosphate by
Phosphoglucomutase.

GLYCOGEN
(n residues)
Pi
Glycogen phosphorylase

GLUCOSE-1-PHOSPHATE

Phosphoglucomutase

GLUCOSE-6-PHOSPHATE

Glucose-6-Phosphatase

GLUCOSE

Diffuse in to the bloodstream

Significance
 Glycogenolysis plays an important role in the fight-or-flight response.
 It contributes to the regulation of glucose levels in the blood.
 The metabolism of glycogen polymers becomes important during fasting.
 In myocytes (muscle cells), glycogen degradation serves to provide an immediate
source of glucose-6-phosphate for glycolysis, to provide energy for muscle
contraction.
 In hepatocytes), the main purpose of the breakdown of glycogen is for the release of
glucose into the bloodstream for uptake by other cells.

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CAM pathway
Pasteur Effect

CAM PATHWAY-

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Pasteur Effect-
 The effect was discovered in 1857 by Louis Pasteur, who showed that aerating
yeasted broth causes yeast cell growth to increase, while
conversely, fermentation rate decreases.
 While the oxygen concentration is low, the product of glycolysis, pyruvate, is turned
into ethanol and carbon dioxide, and the energy production efficiency is low
(2 moles of ATP per mole of glucose).
 If the oxygen concentration grows, pyruvate is converted to acetyl CoA that can be
used in the citric acid cycle, which increases the efficiency to 31 or 29.5 moles of ATP
per mole of glucose. Therefore, about 15 times as much glucose must be consumed
anaerobically as aerobically to yield the same amount of ATP.
 Under anaerobic conditions, the rate of glucose metabolism is faster, but the amount
of ATP produced earlier is smaller.
 When exposed to aerobic conditions, the ATP and Citrate production increases and
the rate of glycolysis slows, because the ATP and citrate produced act as allosteric
inhibitors for phosphofructokinase 1, the third enzyme in the glycolysis pathway.
 The Pasteur effect will only occur if glucose concentrations are low (<2 g/L) and if
other nutrients, mostly nitrogen, are limited.

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Fate of pyruvate- formation of lactate and ethanol
Overview of Calvin cycle
Fate of pyruvate- formation of lactate and ethanol
Overview of Calvin cycle-

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