BIOLOGIC OXIDATION

LEARNING OBJECTIVES
• Explain free energy change, entalphy,entrophy concepts
• Explain endergonic and exergonic reactions
• Count high energy compounds
• Evaluate standard free energy of hydrolysis of
organophosphates of biochemical Importance
• Explain structure of ATP
• Explain reason of high free energy change of hydrolysis of ATP
• Explain importance of phosphagens
• Explain function of myokinase
• Explain structure and function of cytochrome oxidase and
cytochromes
• Explain structure and function of dehydrogenases
• References; Textbook of BIOCHEMISTRY for Medical Students DM Vasudevan, Sreekumari S, Kannan Vaidyanathan,
Harper’s Illustrated Biochemistry, Lippincott’s Illustrated Reviews Biochemistry, Goggle Animation, Medical
Biochemistry J W Baynes, MH Dominiczak, USMLE Road map to biochemistry Macdonald RG and Chaney WG,
RICHARD G. MACDONALD andWILLIAM G. CHANEY
 FREE ENERGY (∆G) AND REDOX POTENTIAL

• Free energy (∆G) is that portion of total energy change in

a system that is available for executing a task,
• It is the actual useful amount of energy, also known as
the chemical potential
• In oxidation and reduction reactions;
• The free energy change is proportionate to the tendency of
reactants to donate or accept electrons.
 If ∆G is negative in sign;
• It means that the products contain less free energy than
the reactants
• Reaction proceeds spontaneously with loss of free energy.
• Reaction is called exergonic,
• Catabolic reactions are exergonic.
 If ∆G is positive in sign;
• It means that the products of the reaction contain more
free energy than the reactants
• Reaction proceeds only if free energy can be gained
• This is known as endergonic reactions
 If ∆G is zero;
• It means that the system is at equilibrium
• No net change takes place.
• Free energy changes are expressed as Redox potential (E’o)
• Increasing negative values of a system have an increasing
tendency to lose electrons.
• Increasingly positive values of a system have an increasing
tendency to accept electrons
• Electrons tend to flow from redox couple to another redox
couple in the direction of the more positive system
• E’o values of various redox couples allow us to predict
the direction of flow of electrons from one redox couple to
another
 Redox potential of redox couples of
components of electron-transport chain
• In practice, an endergonic process cannot exist independently,
• But must be a component of a coupled exergonic-endergonic
system where the overall net change is exergonic.
• Exergonic reactions are termed catabolism (generally, the
breakdown or oxidation of fuel molecules),
• Synthetic reactions that build up substances are termed
anabolism.
• Combined catabolic and anabolic processes constitute
metabolism.
ATP Allows the Coupling of Thermodynamically
Unfavorable Reactions to Favorable Ones
• Endergonic reactions cannot proceed without an input of
free energy.
• Phosphorylation of glucose to glucose-6-phosphate,
the first reaction of glycolysis is highly endergonic and
cannot proceed under physiologic conditions.

• In order to take place, the reaction must be coupled with

another—more exergonic—reaction such as the hydrolysis
of the terminal phosphate of ATP.
ATP Allows the Coupling of Thermodynamically
Unfavorable Reactions to Favorable Ones
• When (1) and (2) are coupled in a reaction catalyzed by
hexokinase, phosphorylation of glucose readily proceeds in
a highly exergonic reaction that under physiologic
conditions is irreversible.
• Many “activation” reactions follow this pattern.
 HIGH-ENERGY PHOSPHATES PLAY A CENTRAL ROLE
IN ENERGY CAPTURE AND TRANSFER
• ATP plays a central role in the transference of free energy
from the exergonic to the endergonic processes.
 ATP= Adenosine Tri Phosphate

• Consists of Nucleoside Adenosine (Adenine linked to

Ribose), and 3 Phosphate groups
 Magnesium is required for
Metabolic function of ATP
• ATP readily forms a complex
with Magnesium ion
• In its reactions in the cell,
ATP functions as the Mg2+
complex
• Mg deficiency impairs virtually
all of metabolism
• ATP contains 2 High-energy Phosphate groups
• ADP contains 1 High-energy Phosphate group
• Phosphate in AMP has ESTER link so has Low-energy
Goggle Animation
 Adenylyl Kinase (=Myokinase)
Interconverts Adenine Nucleotides
• This enzyme is present in most cells.

• Adenylyl kinase is important for the maintenance of energy

homeostasis in cells because it allows:
• 1. Group transfer potential in ADP to be used in the synthesis of ATP.
• 2. AMP formed as a consequence of activating reactions involving
ATP to rephosphorylated to ADP.
• 3. AMP to increase in concentration when ATP becomes depleted
• AMP is able to act as a metabolic (allosteric) signal to increase
the rate of catabolic reactions, which in turn lead to the generation
of more ATP.
 Table 1; Standard Free Energy of Hydrolysis of
Some Organophosphates of Biochemical Importance

Harper’s Illustrated Biochemistry

The Intermediate Value for the Free Energy of Hydrolysis of
ATP Has Important Bioenergetic Significance

• Low- energy phosphates, having a low group transfer

potential are; Ester phosphates found in the intermediates
of glycolysis, have ∆G values smaller than that of ATP
• In high-energy phosphates, with a more negative ∆G ,
the value is higher than that of ATP.
 Table 1; Standard Free Energy of Hydrolysis of
Some Organophosphates of Biochemical Importance

Harper’s Illustrated Biochemistry

 Phosphagens
• Act as storage forms of group transfer potential
• Include creatine phosphate and
• Arginine phosphate, which occurs in invertebrate muscle.
• When ATP is rapidly being utilized as a source of energy for
muscular contraction, phosphagens permit its
concentrations to be maintained,
• When the ATP/ADP ratio is high, their concentration can
increase to act as an energy store.
 Creatine Phosphate
• Act as storage forms of high-energy phosphate
• Provides a high energy reservoir of ATP to regenerate ATP
rapidly
• Occurs in Skeletal muscle, Heart, Spermatozoa,Brain
• Reaction, catalyzed by Creatine Kinase ( = CK )
• ATP + Creatine → Creatine phosphate + ADP
• Reaction is mitochondrial and of special significance in the
myocardium which has a high energy requirement
• Since being a smaller molecule than ATP, Creatine phosphate
can rapidly diffuse from the myocardium to the myofibrils
• The intermediate position of ATP allows it to play
an important role in energy transfer.
• The high free-energy change on hydrolysis of ATP is due to;
• 1.Relief of charge repulsion of adjacent negatively charged
oxygen atoms and
• 2.Stabilization of the reaction products, especially
phosphate, as resonance hybrids.
Free-energy change on hydrolysis of ATP to ADP
• Other “high-energy compounds” are thiol esters involving ;
• Coenzyme A (eg, acetyl- CoA),
• Acyl carrier protein,
• Amino acid esters involved in protein synthesis,
• S-adenosylmethionine ( SAM ),
• Uridine diphosphate glucose, and
• Phosphoribosyl pyrophosphate (PRPP).
ATP ACTS AS THE "ENERGY CURRENCY" OF THE CELL

• The high group transfer potential of ATP enables it to act as

a donor of high-energy phosphate to form those compounds
below in Table 1.
• Likewise, with the necessary enzymes, ADP can accept
phosphate groups to form ATP from those compounds above
ATP in Table 1.
• In effect, an ATP/ADP cycle connects those processes that
generate ~P to those processes that utilize ~P, , continuously
consuming and regenerating ATP.
• This occurs at a very rapid rate since the total ATP/ADP pool is
extremely small and sufficient to maintain an active tissue for
only a few seconds.
 Table 1; Standard Free Energy of Hydrolysis of
Some Organophosphates of Biochemical Importance

Harper’s Illustrated Biochemistry

Role of ATP/ADP cycle in transfer of high- energy phosphate
 Cytochrome Oxidase Is a Hemoprotein
• It is widely distributed in many tissues, having the typical heme
prosthetic group present in myoglobin, hemoglobin, and other
cytochromes.
• It is the terminal component of the chain of respiratory carriers
found in mitochondria and
• Transfers electrons resulting from the oxidation of substrate
molecules by dehydrogenases to their final acceptor, oxygen.
• The action of the enzyme is blocked by carbon monoxide,
cyanide, and hydrogen sulfide.
• This causes poisoning by preventing cellular respiration.
 Cytochrome Oxidases
• Take part in oxidation reduction reactions.
• Contain iron which oscillates between Fe3+ and Fe2+ during the
reactions they participate in.
• Two atoms of copper are present, one associated with each
heme unit.
• Act as electron carriers in the respiratory chain in mitochondria.
• They have an important role in the hydroxylation of steroids in
the endoplasmic reticulum.
• They are NOT dehydrogenase enzymes, although all other
cytochromes are classed as dehydrogenases
 Other Oxidases Are Flavoproteins
• Flavoprotein enzymes contain flavin mononucleotide
(FMN) or flavin adenine dinucleotide (FAD) as prosthetic
groups.
• FMN and FAD are formed in the body from the vitamin
riboflavin.
• FMN and FAD are usually tightly— but not covalently—
bound to their respective apoenzyme proteins.
 DEHYDROGENASES PERFORM 2 MAIN FUNCTIONS
• 1. Transfer of hydrogen from one substrate to another in
a coupled oxidation-reduction reaction;
• These dehydrogenases often utilize common coenzymes or
hydrogen carriers, for example, nicotinamide adenine
dinucleotide (NAD+).

2. Transfer of electrons in the respiratory chain of

electron transport from substrate to oxygen.
 Many Dehydrogenases Depend on
Nicotinamide Coenzymes
• They use NAD+ or nicotinamide adenine dinucleotide
phosphate (NADP+)—or both—which are formed in the
body from the vitamin niacin.
• NADH is generated via the oxidation of fuel molecules.
• NAD+ is regenerated by the oxidation of NADH as
it transfers the electrons to 02 via the respiratory chain in
mitochondria, a process which leads to the formation of
ATP.
• NADP-linked dehydrogenases are found in biosynthetic
pathways where reductive reactions are required.
 Other Dehydrogenases Depend on Riboflavin

• The flavin groups such as FMN and FAD are associated with
dehydrogenases as well as with oxidases.
• The electron acceptor in reactions of the type:

• FAD accepts two electrons and two H+ in the reaction ,

forming FADH2.
 Other Dehydrogenases Depend on Riboflavin

• Flavin groups are generally more tightly bound to their

apoenzymes than are the nicotinamide coenzymes.
• Most of the riboflavin-linked dehydrogenases are
concerned with electron transport in the respiratory chain.
• NADH dehydrogenase acts as a carrier of electrons
between NADH and the components of higher redox
potential.
 Cytochromes May Also Be Regarded as Dehydrogenases

• Cytochromes are iron-containing hemoproteins in which the

iron atom oscillates between Fe3+ and Fe2+ during oxidation
and reduction.
• Except for cytochrome oxidase, they are classified
as dehydrogenases.
• In the respiratory chain, they are involved as carriers of
electrons from flavoproteins on the one hand to cytochrome
oxidase on the other.
• Several identifiable cytochromes occur in the respiratory chain,
that is, cytochromes b, c , c, and cytochrome oxidase aa3.
• Cytochromes are also found in the endoplasmic reticulum
(cytochromes P450 and b5).