Chapter 4 - II

Bioenergetics and biosyntheses

Cellular respiration
• Most living organisms obtain energy by
breaking down organic molecules (catabolism)
during cellular respiration.
• The function of cellular respiration is to
harvest electrons from carbon compounds,
such as glucose, and use that energy to make
Adenosine Tri Phosphate (ATP).
• This catabolic process can be divided into 3 phases.

• Phase I - Breakdown of large complex biomolecules like

polysaccharides, proteins and lipids into their respective
building blocks (hydrolysis).
– The chemical reactions occurring during this stage do not
release much energy. E.g gycolysis
• Phase II - These building blocks are usually oxidized to a
common intermediate, acetyl - CoA.
• Phase III – This consists of the citric acid cycle (i.e. oxidation of
acetyl - CoA to CO2, formation of NADH and FADH2) followed
by electron transport and oxidative phosphorylation.
– Energy released by electron transport to O2 is coupled to ATP
synthesis.
– This cycle is responsible for the release of much energy ( TCA cycle and
ETC).
• Cellular respiration occurs in two main parts:
glycolysis and aerobic respiration.
– The first stage, glycolysis, is an anaerobic process.
– Anaerobic metabolic processes do not require
oxygen.
– Aerobic respiration includes the Krebs cycle and
electron transport chain and is an aerobic process.
• Aerobic metabolic processes require oxygen.
 Glycolysis
Glucose structures
 Glycolysis: anaerobic respiration
• Glucose is a key metabolite in metabolism.
– Glucose is the body’s most readily available source of energy
• Various pathways that are concerned with the utilization, storage, and
regeneration of glucose exist.
– Glycogen is a polymeric storage form of glucose in human and it is
most abundant in the liver and in striated muscle, although some is
found in other tissues also.
• Glycogen is synthesized when glucose supply is high, and its
degradation helps to maintain the blood glucose level when we
are fasting.
• When glycogen is depleted, more glucose is synthesized from
scratch in gluconeogenesis. This pathway’s most important
substrates are amino acids, which are obtained either from a
protein-rich diet-for example, during fasting on meat exclusively-
or, during starvation, from breakdown of cellular protein, mainly in
skeletal muscle.
• Gluconeogenesis occurs in the liver and in the kidneys.
 Glycolysis …
 Greek word- glycos meaning sweet/sugar and
lysis meaning dissolution
 Also known as Embden-Meyerhof ( its
discoverers) Pathway
 Glycolysis, the major pathway for glucose
oxidation, occurs in the cytosol of all cells.
 It is unique, in that it can function either
aerobically or anaerobically, depending on the
availability of oxygen and intact mitochondria.
 It allows tissues to survive in presence or absence of
oxygen, e.g., skeletal muscle.
• The first step in the degradation of glucose is glycolysis, which
breaks down glucose to pyruvate.
• The main purpose of glycolysis is the generation of energy
(ATP). A modest amount of ATP is produced in glycolysis
directly, but much more ATP is formed downstream of
glycolysis through the complete oxidation of pyruvate.
• Glycolysis is the most common pathway for glucose
degradation to pyruvate and is found in animals, plants and
microorganism.
• This pathway is used by anaerobic as well as aerobic
organisms.
• The process takes place in the cytoplasm of prokaryotes and
eukaryotes and does not require oxygen.
• Under aerobic conditions, most of the pyruvate formed in
glycolysis undergoes complete oxidative degradation to CO2
and H2O.
• Pyruvate intended for complete degradation is
transported to the mitochondria, where it is
decarboxylated to acetyl-CoA by pyruvate
dehydrogenase.
• Acetyl-CoA is completely degraded in the citric
acid cycle (or tricarboxylic acid cycle; TCA cycle
for short).
– The “H2” that is produced here is not gaseous but
bound to co-substrates, as NADH and FADH2, which is
subsequently oxidized in the respiratory chain.
 coenzyme A (CoA)

Acetyl-CoA (acetyl coenzyme A)

• If glucose is available in excess of immediate
needs and glycogen is already stocked up to
capacity, it will still be broken down by glycolysis
and pyruvate dehydrogenase to acetyl-CoA.
• However, acetyl-CoA will then not be oxidized,
but it will instead be used for fatty acid synthesis;
the fatty acids are converted to triacylglycerol.
• Fatty acid synthesis occurs in the cytosol of cells
in the liver and fat tissue .
Triglyceride structure
Stages of Glycolysis

Two Stages
A. Preparatory Stage (ATP consuming)
B. ATP producing
• Glycolysis involves ten enzymatic reactions as
described below.
• The first five are preparatory phases or
investment phase.
– Here, glucose is phosphorylated, rearranged and
phosphorylated again, with the two phosphate
groups coming from ATP.
1. The phosphorylation of glucose at carbon-6 by hexokinase
forming glucose 6- phosphate (G6P).
• This reaction consumes ATP, but it acts to keep the glucose
concentration low, promoting continuous transport of glucose
into the cell through the plasma membrane transporters. In
addition, it blocks the glucose from leaking out – the cell lacks
transporters for G6P, and free diffusion out of the cell is
prevented due to the charged nature of G6P. Glucose may
alternatively be formed from the phosphorolysis or hydrolysis
of intracellular starch or glycogen
• In animals, an isozyme of hexokinase called glucokinase is also
used in the liver, has a much lower affinity for glucose, and
differs in regulatory properties. The different substrate affinity
and alternate regulation of this enzyme are a reflection of the
role of the liver in maintaining blood sugar levels.
2. The conversion of glucose-6-phosphate(G6P) to fructose-6-
phosphate(F6P) by phosphohexose isomerase
• The change in structure is an isomerization, in which the
G6P has been converted to F6P. This reaction is freely
reversible under normal cell conditions. However, it is often
driven forward because of a low concentration of F6P,
which is constantly consumed during the next step of
glycolysis. Under conditions of high F6P concentration, this
reaction readily runs in reverse. This phenomenon can be
explained through Le Chatelier's Principle, ''Isomerization
to a keto sugar is necessary for carbanion (negative charge
of carbon that is stable) stabilization in the fourth reaction
step (below)''.
3. The phosphorylation of fructose-6-phosphate to the
1,6-bisphosphate by phosphofructokinase
• The energy expenditure of another ATP in this step is
justified in 2 ways: The glycolytic process (up to this
step) becomes irreversible, and the energy supplied
destabilizes the molecule. Because the reaction
catalyzed by Phosphofructokinase 1 (PFK-1) is coupled
to the hydrolysis of ATP (an energetically favorable
step) it is, in essence, irreversible, and a different
pathway must be used to do the reverse conversion
during gluconeogenesis. This makes the reaction a key
regulatory point. This is also the rate-limiting step.
• The second phosphorylation event is necessary to
allow the formation of two charged groups (rather than
only one) in the subsequent step of glycolysis, ensuring
the prevention of free diffusion of substrates out of the
cell.
• The same reaction can also be catalyzed by
pyrophosphate-dependent phosphofructokinase (PFP
or PPi-PFK), which is found in most plants, some
bacteria, archea, and protists, but not in animals. This
enzyme uses pyrophosphate (PPi) as a phosphate
donor instead of ATP. It is a reversible reaction,
increasing the flexibility of glycolytic metabolism.
4. The cleavage of fructose-1,6-bisphosphate by aldolase.
– This yields two different products, dihydroxyacetone phosphate and
glyceraldehyde-3-phosphate,
• Destabilizing the molecule in the previous reaction allows the
hexose ring to be split by aldolase into two triose sugars:
dihydroxyacetone phosphate (a ketose), and glyceraldehyde 3-
phosphate (an aldose). There are two classes of aldolases: class I
aldolases, present in animals and plants, and class II aldolases,
present in fungi and bacteria; the two classes use different
mechanisms in cleaving the ketose ring.
• Electrons delocalized in the carbon-carbon bond cleavage associate
with the alcohol group. The resulting carbanion is stabilized by the
structure of the carbanion itself via resonance charge distribution
and by the presence of a charged ion prosthetic group.
5. The isomerization of dihydroxyacetone
phosphate to a second molecule of glyceraldehyde-
3-phosphate by triose phosphate isomerase
• Triosephosphate isomerase rapidly interconvert
dihydroxyacetone phosphate with glyceraldehyde
3-phosphate (GADP) that proceeds further into
glycolysis. This is advantageous, as it directs
dihydroxyacetone phosphate down the same
pathway as glyceraldehyde 3-phosphate,
simplifying regulation
6. The dehydrogenation and concomitant phosphorylation of
glyceraldehyde-3-phosphate to 1,3-bis-phosphoglycerate by
glyceraldehyde-3-phosphate dehydrogenase
• The aldehyde groups of the triose sugars are oxidised, and
inorganic phosphate is added to them, forming 1, 3-
bisphosphoglycerate.
• The hydrogen is used to reduce two molecules of NADH, a
hydrogen carrier, to give NADH + H+ for each triose.
• Hydrogen atom balance and charge balance are both
maintained because the phosphate (Pi) group actually
exists in the form of a hydrogen phosphate anion (HPO42- ),
which dissociates to contribute the extra H ion and gives a
net charge of -3 on both sides.
7. The transfer of the 1-phosphate group from 1,3-bis-
phosphoglycerate to ADP by phosphoglycerate kinase, which
yields ATP and 3-phosphoglycerate.
• At this step, glycolysis has reached the break-even point: 2
molecules of ATP were consumed, and 2 new molecules
have now been synthesized. This step, one of the two
substrate-level phosphorylation steps, requires ADP; thus,
when the cell has plenty of ATP (and little ADP), this
reaction does not occur. Because ATP decays relatively
quickly when it is not metabolized, this is an important
regulatory point in the glycolytic pathway.
• ADP actually exists as ADPMg− , and ATP as ATPMg2−,
balancing the charges at -5 both sides.
8. The isomerization of 3-phosphoglycerate to 2-phosphoglycerate by
phosphoglycerate mutase (PGAM)

9. The dehydration of 2-phosphoglycerate to phosphoenolpyruvate by

enolase.

10. The transfer of the phosphate group from phosphoenolpyruvate to

ADP by pyruvate kinase, to yield a second molecule of ATP.
• A final substrate-level phosphorylation now forms a molecule of
pyruvate and a molecule of ATP by means of the enzyme pyruvate
kinase. This serves as an additional regulatory step, similar to the
phosphoglycerate kinase step.
Glycolysis steps
Glycolysis
Regulation (glycolysis)
• Three reactions physiologically irreversible
• These are the major sites of regulation.
i. Hexokinase / Glucokinase
ii. Phosphofructokinase Key regulatory enzymes
iii. Pyruvate Kinase
Biochemical logic for the presence of regulatory steps
• The existence of more than one point of regulation
indicates that intermediates between those points enter
and leave the glycolysis pathway by other processes. For
example, in the first regulated step, hexokinase converts
glucose into glucose-6-phosphate. Instead of continuing
through the glycolysis pathway, this intermediate can be
converted into glucose storage molecules, such as glycogen
or starch. The reverse reaction, breaking down, e.g.,
glycogen, produces mainly glucose-6-phosphate; very little
free glucose is formed in the reaction. The glucose-6-
phosphate so produced can enter glycolysis after the first
control point.
• In the second regulated step (the third step of
glycolysis), phosphofructokinase converts fructose-6-
phosphate into fructose-1, 6-bisphosphate, which then
is converted into glyceraldehyde-3-phosphate and
dihydroxyacetone phosphate. The dihydroxyacetone
phosphate can be removed from glycolysis by
conversion into glycerol-3-phosphate, which can be
used to form triglycerides. Conversely, triglycerides can
be broken down into fatty acids and glycerol; the latter,
in turn, can be converted into dihydroxyacetone
phosphate, which can enter glycolysis after the second
control point.
Overall Reaction (of glycolysis)

Aerobic
Glc + 2NAD+ + 2ADP + 2Pi

2 pyruvate + 2NADH + 2H+ + 2ATP + 2H2O

Glc + 2ADP + 2Pi 2 Lactate + 2ATP + 2H2O

Anaerobic
Glycolysis - Summary

Glucose Glucose
2 ATP
4 ADP
2 ADP 2 ATP
4 ADP
4 ATP
2 ADP
2 NAD 4 ATP

2 NADH + H

2 Pyruvate 2 Lactate
Bioenergetics
• AEROBIC GLYCOLYSIS

2NADH + H+ = 2 x 2.5ATP = 5 ATP

Substrate Level = 2 x 2 ATP = 4 ATP

_____________________________________

TOTAL = 9 ATP

Consumed = -2 ATP
_____________________________________

Net Balance = 7 ATP

Bioenergetics
• ANAEROBIC GLYCOLYSIS (e.g. RBC)
Substrate Level = 4 ATP
Consumed = -2 ATP
____________________________
Net Balance = 2 ATP
Entry of other hexoses into glycolysis
Galactose
Galactokinase
Galactose Gal-1-Phosphate
Mg+2
ATP ADP
UDP-Glc
Gal-1-P-uridyl UDP-Gal-4-epimerase
transferase
UDP-Gal

Glc-1-Phosphate

Mutase

Glc-6-Phosphate Glycolysis
Fructose (Major pathway) (Liver)
ATP ADP Fr-1-P
Mg++ Aldolase
Fructose Fr-1-Phosphate DHAP + Glyceraldehyde
Fructokinase
ATP
Triose
PTI ADP Kinase

Glycolysis Glyceraldehyde -3-Phosphate

Fructose (Minor pathway) (Liver)

ATP ADP
Mg++
Fructose Fr-6-Phosphate  Glycolysis
Hexokinase
Thank You!