LEARNING UNIT 1

The importance of energy changes and

electron transfer in Metabolism

Outcome 3: “High-energy” compounds

References
➢ WILLEY, J.M., SHERWOOD, L.M. & WOOLVERTON, C.J. (2011) Prescott’s Microbiology. 8th ed.
Singapore. McGraw-Hill.
➢ BOYER, R. (2006) Concepts in Biochemistry. 3rd ed. USA: John Wiley & Sons, Inc.
➢ KIM, B.H. & GADD, G.M. (2008) Bacterial Physiology and Metabolism. Cambridge University Press
➢ COHEN, G.N. (2004) Microbial Biochemistry. 2nd ed. Springer.
➢ VOET, D., VOET, J.G. & PRATT, C.W. (2006) Fundamentals of Biochemistry. 2de edition, John
Wiley & Sons, USA

Learning objectives

• Explain the meaning of “high-energy” compounds.

• Demonstrate knowledge in the role of ATP in transferring energy from exergonic
processes to endergonic processes.
• Discuss several factors that contribute to the release of large amounts of free energy
from high-energy compounds.
• Understand that electrons released in oxidative processes can be transferred to
cofactors.
• Discuss several coupled reactions in metabolism.
• Calculate the free energy change for reactions.

1. WHAT ARE “HIGH-ENERGY” COMPOUNDS?

High-energy metabolites are molecules that contain chemical bonds with high potential
energy, which can be released and harnessed to perform energy-requiring cellular processes.
These molecules are typically involved in metabolic pathways that produce energy, such as
cellular respiration, and are important for the normal functioning of living cells.

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Energy-rich or high-energy compounds exhibit a large decrease in free energy

(ΔG°’> -25 kJ.mol-1) when they undergo hydrolytic reactions. Generally, these compounds
contain high-energy phosphate bonds or other unstable chemical bonds that are easily
hydrolyzed, releasing energy. They are in unstable to acid, alkali, and heat.

2. WHAT ARE “COUPLED” REACTIONS?

Coupled reactions refer to the biochemical reactions in which the energy released by one
reaction (exergonic reaction) is used to drive an energy-requiring reaction (endergonic
reaction). These reactions are important for the efficient use of energy in living cells, as they
allow metabolic pathways to be tightly regulated and interconnected.

One of the most common examples of coupled reactions is the hydrolysis of adenosine
triphosphate (ATP) to adenosine diphosphate (ADP) and inorganic phosphate. The energy
released by this reaction can be used to drive other energy-requiring processes.

Figure 1: Coupling of reactions

(https://www.google.com/search?q=coupling+reactions+delta+g&sxsrf=AJOqlzULGPoWPGUL_R7dJ9E27Sd8RNU6pg:1678
788250010&source=lnms&tbm=isch&sa=X&ved=2ahUKEwj0363Kldv9AhUpSEEAHfBzD7sQ0pQJegQIBhAC&biw=1536&bih
=730&dpr=1.25#imgrc=D59oCGvtYfdsjM)

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Coupled reactions can also involve the transfer of electrons or other chemical groups
between molecules. For example, the transfer of electrons from nicotinamide adenine
dinucleotide (NADH) to oxygen in the electron transport chain is coupled to the synthesis of
ATP from ADP and inorganic phosphate. This coupling allows the energy released by the
transfer of electrons to be used to drive ATP synthesis (This will be discussed later).

3. CLASSES OF HIGH-ENERGY COMPOUNDS

High-energy compounds can be broadly classified into three main classes:

1. Phosphorylated compounds: These are compounds that contain high-energy

phosphate bonds, which can be broken to release energy. Various types of
phosphorylated compounds include:

Table 1: High-energy phosphorylated compounds

Types of high-energy phosphorylated Examples

compounds
Pyrophosphates ATP
Enol phosphates Phosphoenolpyruvate
Acylphosphates 1,3 Bisphosphoglycerate
Guanido phosphates (phosphagens) Phosphocreatine

2. High-energy compounds with thioester bonds (Thioesters): These are compounds

that contain high-energy thioester bonds, which can be hydrolyzed to release energy.
Examples of high-energy compounds with thioester bonds include acetyl-coenzyme A
(acetyl-CoA) and S-adenosylmethionine (SAM).

3. Reducing agents: These are compounds that can donate electrons to other
molecules, releasing energy in the process. Examples of reducing agents include:

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a) nicotinamide adenine dinucleotide (NADH),

b) nicotinamide adenine dinucleotide phosphate (NADPH),
c) flavin adenine dinucleotide (FADH2), and
d) reduced coenzyme Q.

All three classes of high-energy compounds play important roles in various metabolic
pathways and are essential for the normal functioning of living cells. They can be used to
transfer and store energy, drive energy-requiring processes, and perform various biochemical
reactions. Disruptions in the metabolism of high-energy compounds can lead to various
diseases and disorders.

4. PHOSPHORYL GROUP TRANSFER POTENTIAL

The phosphoryl group transfer potential is a measure of the tendency of phosphorylated

compounds to transfer their phosphoryl group (PO32-) to water. As previously mentioned, a
compound is considered to be high-energy when it exhibits a large decrease in free energy
(ΔG°’> -25 kJ.mol-1) when hydrolyzed.

Table 2: Several high-energy phosphorylated compounds

High-energy
compounds

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5. EXPLAINING THE HIGH-ENERGY NATURE OF PHOSPHORYLATED COMPOUNDS

5.1. PYROPHOSPHATE COMPOUNDS

ATP and GTP are examples of pyrophosphate compounds. Pyrophosphate compounds are
high-energy metabolites because they contain a pyrophosphate group (-PPi) that has a high-
energy phosphoanhydride bond(s). The large negative ΔG°’ for the hydrolysis of the
phosphoandride bond(s) indicated that the reactants contain more free energy than the
products. This energy difference can be explained by:

1) The greater like-charge repulsion (electrostatic repulsion) between the four negative
charges in an ATP molecule compared to that in an ADP molecule with only three
negative changes, contributes significantly to the free energy content in the molecule.
In addition, there will be a tendency for the electrons in the P=O bond of the
phosphates to be drawn closer to the electronegative oxygen atom, thereby producing
a partial negative charge (-) on the oxygen atom, increasing the electrostatic repulsion
within the molecule. This like-charge repulsion creates an unstable state, destabilizes
ATP, which makes the pyrophosphate bond highly reactive and energetically favorable
to break.

Figure 2: Structure of ATP

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Hydrolysis of the terminal phosphate of ATP is called orthophosphate cleavage. Hydrolysis

of the interior pyrophosphate bond of ATP is called pyrophosphate cleavage.

Figure 3: Hydrolysis of ATP

Several factors influence the electrostatic repulsion within a molecule:

a) pH
The amount of electrostatic repulsion is affected by pH. The reactants and products of ATP
hydrolysis have ionizing functional groups with pka values between 6 and 7. Changes in pH
alter the ionic states of all reactants and products in the reaction. At high pH, when the
ionizing groups are relatively dissociated, electrostatic repulsion increased.

b) Cellular concentration of magnesium ions (Mg2+)

All of the ATP and ADP molecules in a cell exist in complexes with Mg 2+. Hydrolysis of
MgATP is less favorable than hydrolysis of uncomplexed ATP. Mg2+ partially neutralizes the
negative charges on the oxygen atoms of ATP, thereby diminishing the electrostatic
repulsion.

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2) The resonance stabilization of the hydrolyzed products of ATP also contributes to the
large decrease in the free energy when ATP is hydrolyzed. ADP and Pi have greater
resonance stabilization compared to ATP. For example, at pH 7, a free phosphate
group can exist in more resonance forms than a phosphate group tied up in an ester or
anhydride bond. As a result, the release of a phosphate group through the hydrolysis
of a phosphoanhydride bond in ATP leads to an increase in the number of resonance
structures and a decrease on the free energy.

Figure 4: Resonance structures of orthophosphate.

3) The interaction between a solvent and solute also influences the free energy content of
the molecules involved. The solvation energy of a compound is the energy released
because of the noncovalent interactions between the compound and the solvent
molecules. For example, water can bind more effectively to the hydrolysis products of
ATP than ATP itself. When ions are solvated, they are electrically shielded from each
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other, decreasing the electrostatic repulsion between the phosphate groups. In other
words, the solvation energy for ATP is lower than for its hydrolysis products.

5.2. ENOYL PHOSPHATES

Phosphoenolpyruvate is an example of an enoyl phosphate. The large negative ΔG°’ for the
hydrolysis of phosphoenolpyruvate can be explained by the resonance stabilization of the
released phosphate group (explained above) and the spontaneous conversion of the enol
form of a ketose to a more stable keto form.

Figure 5: Hydrolysis of phosphoenolpyruvate

Upon hydrolysis of phosphoenolpyruvate the enol form of pyruvate is formed. The enol form
of pyruvate can immediately tautomerize to the more stable keto form. Because
phosphoenolpyruvate has only one tautomeric form (enol) and the product, pyruvate, has two
possible forms, pyruvate is more stabilized than phosphoenolpyruvate. This enol-keto

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tautomerization is responsible for making phosphoenolpyruvate one of the most “energy-rich”

phosphate compounds.

5.3. ACYL PHOSPHATES

1,3-Bisphosphoglycerate and acetyl phosphate are examples of acyl phosphates. Acyl

phosphate compounds are high-energy molecules that contain a carboxylate group (-COO-)
and a phosphoryl group (-PO3 2-). The bond between the carboxylate group and the
phosphoryl group is a high-energy phosphoanhydride bond, similar to the bond found in ATP.
This bond has a large amount of potential energy that can be released when it is hydrolyzed,
providing energy for cellular processes.

Figure 6: Basic structure of an acyl phosphate

Electrostatic repulsion in acyl phosphate is a significant factor contributing to the large

negative standard free energy of hydrolysis of this class of compounds. The C=O bond of the
acyl phosphate group has considerable polar character because of the tendency for the
electrons in the double bond to be drawn closer to the electronegative oxygen. Energy is
required to overcome the repulsion between the partial positive charges on the carbon and
phosphorus atoms, such energy being released on hydrolysis of the acyl phosphate.

The amount of free energy released during the hydrolysis of an acyl phosphate compound
depends on the nature of the molecule that is formed after hydrolysis. The more stable the
molecule, the more free energy is released. For example, the hydrolysis of acetyl phosphate
produces acetate and inorganic phosphate. Acetate is a stable molecule that does not readily

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react with other molecules, so a large amount of free energy is released during the hydrolysis
of acetyl phosphate. This free energy can be used to drive other energy-requiring processes
in the cell.

Figure 7: Hydrolysis of acetyl phosphate

The hydrolysis of 1,3-bisphosphoglycerate results in the formation of 3-phosphoglyceric acid

which can immediately lose a proton to form the carboxylate ion (3-phosphoglycerate) which
has two equally probable resonance forms.

Figure 8: Hydrolysis of 1,3-bisphosphoglycerate (http://kocw-

n.xcache.kinxcdn.com/data/document/2018/pusan/mindosik1118/02.pdf)

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5.4. PHOSPHAGENS

Another type of energy-rich compound that plays an important role in energy transfer and
storage is the phosphagens. This type of structure is found in phosphocreatine and
phosphoarginine. Phosphagens are phosphoamides and is found in animal muscle cells. In
the muscles of vertebrates, large amounts of phosphocreatine are formed during times of
ample ATP supply. When ATP levels fall, creatine kinase catalyzes rapid replenishment of
ATP though transfer of the activated phosphoryl group form phosphocreatine to ADP.

Figure 9: Hydrolysis of phosphocreatine

6. THIOESTERS

Thioesters are high-energy molecules that contain a sulfur atom in place of the oxygen atom
found in esters. Thioesters are important intermediates in many metabolic pathways,
including fatty acid metabolism and the citric acid cycle. Examples of important thioesters
include acetyl-CoA and succinyl-CoA.

The amount of free energy released during the hydrolysis of a thioester bond is dependent on
the nature of the molecule that is formed after hydrolysis. In the case of acetyl-CoA the bond
is hydrolyzed to produce acetate and CoA. Acetate is a more stable molecule than acetyl-
CoA, so a large amount of free energy is released during the hydrolysis of the thioester bond.
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Figure 10: Hydrolysis of acetyl-CoA

The standard free energy change (ΔG°') for the hydrolysis of acetyl-CoA is -31.5 kJ/mol,
which is slightly higher than the ΔG°' for the hydrolysis of ATP (-30.5 kJ/mol).

In summary, the products of high-energy compounds are more stable than the reactants by
one or more of the following:
1) charge separation relieves electrostatic repulsion,
2) by resonance stabilization,
3) by isomerization (tautomerization)
4) by lower solvation energies.

7. REDUCING AGENTS

Oxidation-reduction reactions are very common in metabolism. It involves electron transfer

from one atom or molecule to another atom or molecule. An oxidizing agent is a substance
that causes oxidation (loss of electron(s)). It is the substance that accepts electron(s) or
removes hydrogen atoms from another compound, increasing the oxidation state of that
compound.

A reducing agent is a substance that causes reduction (gain of electron(s)). It is a substance

that donates electron(s) or adds hydrogen atoms to another compound, decreasing the
oxidation state of that compound.

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Figure 11: Transfer of electrons (https://byjus.com/chemistry/oxidizing-agent/)

Reducing agents are high-energy compounds that play an important role in many biochemical
processes. These compounds contain electrons that can be donated to other molecules,
reducing them and transferring energy in the process.

Nicotinamide adenine dinucleotide (NADH) and flavin adenine dinucleotide (FADH 2) are two
examples of reducing agents. These coenzymes are involved in cellular respiration, a
metabolic process that produces ATP from glucose. During cellular respiration, NADH and
FADH2 donate electrons to the electron transport chain, which is a series of proteins that
generate a proton gradient across a membrane. This gradient is then used to produce ATP.

Figure 12: Electron transport chain

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Figure 13: Nicotinamide adenine dinucleotide

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Figure 14: Flavin adenine dinucleotide.
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Most reduced

Figure 15: Oxidative state of carbon Most oxidised

8. WHAT IS SUBSTRATE-LEVEL PHOSPHORYLATION?

Substrate-level phosphorylation is a type of phosphorylation that involves the transfer of a

phosphate group from a high-energy molecule, such as a phosphorylated intermediate, to
ADP, producing ATP. A variety of specific kinases catalyze these reactions. The transfer of
the activated phosphoryl group from acetyl phosphate to ADP provides lactobacilli with ATP
needed for anaerobic metabolism. Substrate-level phosphorylation occurs in the cytoplasm
during glycolysis and in the mitochondrial matrix during the citric acid cycle.

O
-
HO P O
O-
O O Acetate
H2O kinase
-
CH3 C O P O O
- -
O CH 3 C O
Acetylphosphate ADP Acetate
ATP

Figure 16: Substrate-level phosphorylation catalyzed by acetate kinase.

The standard free energy of hydrolysis of acetyl phosphate is greater than that of the
hydrolysis of ATP (-42.2 kJ.mol-1). Thus, under standard conditions the hydrolysis of acetyl
phosphate can be coupled to the synthesis of ATP.

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