College of Health Sciences

Department of Biomedical Science

Biochemistry for Medical Students (Respiratory Module)

By Melaku Mekonnen

June 2023

1
Biochemistry of Respiration

2
 Outlines

Acid-Base Regulation

Hemoglobin: structure and function (transport of O 2 and CO2)

Lung Surfactants

3
 Introduction
In biochemistry, respiration (both in animal and plant cells) refers to the series of
biochemical oxidations in which organic molecules are converted to CO2 and H2O to
provide energy.
Breathing is a physical process while respiration is a biochemical process.

The chemical energy obtained from respiration is trapped in the cells.

Respiratory chemistry refers to the balance of O2 and CO2.

CO2 is far from a waste gas and is important in maintaining pH, electrolyte

balance, and O2 distribution.

In fact, breathing is derived from CO2 levels. 4

 Cont’d…
The main functions of the respiratory system are:

to obtain O2 from the external environment and supply it to the cells

to remove CO2 produced by cellular metabolism from the body.

There are different factors affecting respiration:

Chlorine, Bromine, and Ozone affect the upper and lower region of the
respiratory tract.

Nitrogen Dioxide (NO2), phosgene (COCl2), and arsenic trichloride affect the
terminal regions of the lung.

How do these factors affect respiration???? 5

 Acids and Bases
According to Bronsted’s concept, acids are defined as substances that are capable
of donating a proton (H+ ion).

On ionization, acids donate a proton and at the same time, a corresponding base
that is capable of accepting a proton is formed. Such a base is called a conjugate
base.

Acids are distinguished by their sour taste. Some common examples are acetic acid
in vinegar, citric acid in citrus fruits, and lactic acid in sour milk products.
Acid (HA) H+ + conjúgate base (A-)

6
 Cont’d.…
Based on their dissociation, acids are classified into two groups (strong and weak acids).

Strong acids are those which dissociate completely in solutions so that the concentration of H+ is
the same as the number of hydrogen atoms in the acid. They donate large numbers of hydrogen
ions to a solution.

E.g. HCL H + + Cl-

H2SO4 2H + + SO4-

Weak acids are those which do not completely dissociate and the concentration of H+ depends
on the dissociation constant of the acid (based on the extent of dissociation).

E.g. CH3 COOH CH3COO - + H+

H2 CO3 HCO 3- + H+ 7
 Cont’d...

 Besides, acids can be classified into two classes (mineral and organic acids).

Mineral acids are inorganic compounds.

 Examples: HCl, H2SO4, HNO3

Organic acids are organic compounds and contain an acidic carboxylic group (-COOH).
 Examples: Acetic, citric, fumaric, malic, pyruvic, glutaric, lactic, and succinic
acids.
 Organic acids are weak acids & many of them are found as intermediates in the
metabolic pathways of the body.
 FAs can be considered as organic acids, but are often included under the class of lipids.
8

 Cont’d…
Likewise, according to Bronsted’s concept, bases are substances that are capable of accepting a
proton (H+ ion).

Bases are classified into strong and weak bases.

Strong bases can dissociate completely (100% dissociation).

The number of OH- ions formed from dissociation will be equal to the number of base molecules.

 NaOH Na + + OH-
 KOH K + + OH-

Weak bases are those whose dissociation is not complete.

 Ammonium hydroxide
9
 pH

 pH of a solution is defined as the negative base 10 logarithms of its H+ ion

concentration (PH = -log H+ ). OR
 It is the logarithm of the reciprocal of OH- ion concentration (PH = log 1/
H+ ).
 The pH scale indicates the number of hydrogen ions in a solution and indicates
the relative acidity of a solution and ranges from 0 to 14.

10
 Cont..
 A solution having pH 7 is called a neutral solution where the number of H+ ions is equal to
the number of OH - ions.
 Pure water will have a pH of 7.

A solution having a pH less than 7 is called an acidic solution where the number of H+ ions
will be higher relative to the number of OH- ions.

A solution having a pH above 7 is called a basic solution where the number of OH- ions
will be higher relative to the number of H+ ions.

11
 Cont’d…

The pH decreases as the H+ concentration increases.

The pH increases as the OH- concentration increases.

The normal value of pH in blood is 7.35 to 7.45.

 Acidosis (acidemia): below 7.35
 Alkalosis (alkalemia): above 7.45
Death occurs: when < 6.8 or > 8.0

12
Cont..

13
 Physiological pH
 The normal PH of the body is maintained at 7.4 which is called physiological pH.

 Physiological pH is the optimum pH at which most of the biochemical reactions in the

body take place at maximum speed.

 Since any deviation from the normal physiological pH may lead to death,

maintenance of normal physiological pH is essential for the maintenance of life.

Acidosis
It is a condition in which the body pH is decreased below the normal range.

Acidosis is caused either by an increase in H+ concentration or by a decrease in OH-

concentration. 14
 Cont’d...
Alkalosis
It is a condition in which the pH is increased beyond the normal range.

Alkalosis is caused either by a decrease in the concentration of H+ ions or by an

increase in the concentration of OH- ions.

 Both acidosis and alkalosis:

affect the activity of enzymes

affect the speed of biochemical reaction

leading to metabolic imbalance and death.

15
 Acid-Base Balance

Acid-base balance refers to the balance between input (intake and production)
and output (elimination of hydrogen ion).

The lungs are able to prevent an increase in the Pco2 in the blood by excreting

the CO2 produced by the body since normal acid-base balance can be
maintained by the lungs in addition to that of the kidneys.

CO2 is a weak acid and a by-product of normal metabolism.

16
 Cont’d…

Acid-base balance of the body is the body's pH balance and the level
of acids and bases in the blood at which the body functions best
(properly).

The human body is built to naturally maintain a healthy balance of

acidity and alkalinity by using the two organ systems (lungs and
kidneys) which play a key role in this process.

17
Cont’d…

18
 Importance of Maintenance of Acid-Base Balance
Maintenance of acid-base balance is fundamental for:

the normal functioning of biological processes.

 This is mainly due to the fact that the activities and functions of many enzymes
are pH dependent.

Any disturbance in the pH range affects multi-systems and processes in the body including:

 Oxygenation

Enzymes

Metabolic functions

Systems like muscles and nerve system

19
Cont’d…

20
Cont’d…

21
 Regulation of Acid-Base Balance by the Lungs

The body uses different mechanisms to control the blood’s acid-base balance.

There are three important mechanisms that the body uses to regulate pH.

 chemical buffer (first line of defense)

 respiratory system (second line of defense)

 urinary system (second line of defense)

These three mechanisms work together to keep body pH within a narrow range.

22
 Cont’d…

One of the mechanisms by which the body uses to control blood pH

involves the release of CO2 by the lungs.

The respiratory system contributes to the balance of acids and bases in the
body by regulating the levels of CO2 in the blood.

The lungs ensure the removal of carbonic acid in the form of CO2.

23
 Cont’d…

CO2 is a mildly acidic waste product of the metabolism of O2 and nutrients (which

all cells need).

 It is constantly produced by the cells and then passes from the cells into the
blood.

The blood carries CO2 to the lungs, where it is exhaled.

As CO2 abnormally accumulates in the blood, the pH of the blood decreases

(acidity increases).
24
 Cont’d…

The brain regulates the amount of CO2 that is exhaled by controlling the speed and

depth of breathing (ventilation).

The amount of CO2 exhaled, and consequently, the pH of the blood increases as

breathing becomes faster and deeper.

Therefore, the brain and lungs are able to regulate the blood pH by adjusting the
speed and depth of breathing.

The mechanism by which the brain regulates the speed and depth of breathing????
25
Cont’d…

26
 Cont’d…

The lungs (respiratory system) are the major organs (systems) that control
plasma pH by adjusting the concentration of CO2.

The equilibrium between dissolved CO2 and H2CO3 is accelerated by carbonic

anhydrase.

Here, respiratory alkalosis results from hyperventilation as the primary

disturbance.

Hyperventilation also forms the respiratory compensation of metabolic

27
acidosis.
 Cont’d…

The increase in pH is often caused by hyperventilation or excessively deep

breathing.

Respiratory alkalosis occurs when you breathe too fast or too deep and CO2
levels drop too low.

When we hyperventilate, we exhale more CO2 than normal.

As a result, the CO2 concentration in the blood is reduced and the increased
bicarbonate level is present as compared to carbonic acid.
28

 Cont’d…

The body compensates for both alkalosis and acidosis mainly through the lungs.

The lungs change the alkalinity of the blood by allowing more or less carbon
dioxide to escape while breathing.

Respiratory compensation for metabolic acidosis increases the respiratory rate

to drive off CO2 and readjust the bicarbonate to carbonic acid ratio to the
20:1 level.

This adjustment can occur within minutes. However, it is not as proficient as its
compensation for acidosis
29
 Respiratory Response to Metabolic Acidosis

Respiratory Compensation (Alkalosis)

A primary metabolic acidosis stimulates peripheral chemoreceptors

(which respond to low pH or high H+), causing:
hyperventilation

 decrease in pCO2

secondary respiratory alkalosis

30
 Cont’d…

o The bicarbonate buffer system has two components that include plasma [CO2]

and [HCO3−].

o The respiratory system controls plasma pH by adjusting the [CO2].

o The equilibrium between dissolved CO2 and H2CO3 is enhanced by carbonic

anhydrase.

o Respiratory alkalosis has been resulted from hyperventilation as the primary

disturbance.
31
Cont’d…

32
The Role of Respiratory System in Maintaining the Acid-Base Balance

Body produces acids in the form of hydrogen ions.

It’s important to neutralize it to maintain normal internal homeostasis

of the body for optimal functions of enzymes.

There are many buffers in the body and 2 of the most important and
additional secondary defenses are being respiratory and renal systems.

Body cells can also utilize oxygen and produce acids or hydrogen ion.

33
 Cont’d…

This oxygen is delivered to the cells by RBCs which pick it up from the lungs.

So basically RBCs act as a transport system between lungs and tissues.

The same RBCs after giving oxygen pick up hydrogen ions from tissues by
combining them with bicarbonate ions to form CO2 and water.

RBCs then bring this CO2 back to the lungs where they release it and take up
oxygen.

34
 Cont’d…
This is the normal process in the case of increased metabolism by tissues

for example during exercise or fever, the heart pumps more blood to tissues

to deliver more oxygen and collect more hydrogen ions produced.

Similarly there is increased breathing in parallel to provide more oxygen

and remove the CO2 delivered to lungs.

• Thus by clearing CO2, lungs help in maintaining body’s acid-base balance.

35
 Cont’d…
In order to operate effectively, blood must possess the appropriate
combination of acid and base molecules.

The physiological modulation of the pH of an organism's natural interstitial

liquid is commonly characterized as acid-base maintenance.

The level of CO2 gas emitted from the body system is mainly evaluated by
measuring the level of gasses taken or expelled by the individual.

36
 Buffer Systems
Another mechanism for controlling acid-base balance involves the use of chemical
buffer systems, which guard against sudden shifts in acidity and alkalinity.

The pH buffer systems are combinations of the body's own naturally occurring weak
acids and weak bases.

These weak acids and bases exist in pairs that are balanced under normal pH
conditions.

The pH buffer systems work chemically to minimize changes in the pH of a solution

by adjusting the proportion of acid and base.

37
 Cont’d…
Bicarbonate Buffer System

Bicarbonate buffer is the most important buffer system in blood plasma (generally
in the extracellular fluid) which accounts for about 65% of plasma buffering
capacity.

It consists of weak acid H2CO3 (pK1 = 6) and conjugated base HCO3– (bicarbonate).

It operates by interconversion of the two components (carbonic acid and bicarbonate).

38
39
 Acid-Base Imbalances
 Acid-base regulation is maintained by the two active and specialized organs to
maintain the acid-base balance in the body (kidneys and lungs).

 Even slight variations from the normal range can have significant effects on different
vital organs so the two organs must work their maintenance activity properly.

 The blood's acid-base balance must be precisely controlled to prevent the severe
effects of acid-base imbalance on many organs.

 Acid–base balance is disturbed due to metabolic, respiratory, and renal disorders.

40
 Cont’d…
 Arterial blood gas analysis is a test most frequently done on critically ill patients
in the emergency, recovery, and intensive care units.

 The main purposes of this test is an assessment of acid-base status, which is

disturbed during critical illness.

 There are four simple acid-base disorders:

1. metabolic acidosis
2. respiratory acidosis
3. metabolic alkalosis
4. respiratory alkalosis 41
 Cont’d…

Supressed and enhanced ventilation, involving retention and depletion of

carbon dioxide, results in respiratory acidosis and respiratory alkalosis,
respectively.

Respiratory acidosis and respiratory alkalosis are caused by changes in CO2

exhalation due to lung or breathing disorders

Metabolic acidosis & metabolic alkalosis are caused by an imbalance in the

production of acids or bases and their excretion by the kidneys.

42
 Cont’d…

Metabolic Alkalosis: is caused by a loss of hydrogen ions due to different reasons

like vomiting accompanied by a loss of hydrochloric acid, with a rise in blood
plasma bicarbonate along with a decrease in other anions (principally chloride) or
an increase in cationic sodium.

 These alterations are immediately reflected in blood pH as alkalemia.

43
 Cont’d…
Metabolic Acidosis: is caused by the retention of hydrogen ions or a loss of bicarbonate.

 Excess production of ketoacids due to DM or starvation, or lactate in hypoxia is

accompanied by an increase of hydrogen ions which consumes bicarbonate in the buffering
system.

 This condition can also be enhanced by a loss of CO2 in respiratory compensation involving
hyperventilation.

 Renal insufficiency also results in failure in the excretion of hydrogen ions and
reabsorption of bicarbonate.

 A loss of alkaline secretion in the intestines due to diarrheal disease also develops into
acidosis due to a loss of bicarbonate, superimposing water, and sodium depletion.
44
 Cont’d..

 Metabolic acidosis is the most common disorder encountered in clinical

practice.
It is a potent stimulator of NaCl absorption, whereas metabolic alkalosis
inhibits this process.

45
 Anion Gap
The total concentration of cations and anions which is expressed as
milliequivalents per liter (mEq/l) is equal in the body fluids.
This is required to maintain electrical neutrality.

The commonly measured electrolytes cations and anions in the plasma include:

Na+, K+, Cl- and HCO3-

Na+ and K+ together constitute about 95% of the plasma cations.

Cl- and HCO3- are the major anions, contributing to about 80% of the plasma
anions.
46
 Cont’d...

Anion gap is defined as the difference between the total concentration of

cations (Na+ and K+) & that of the anion (Cl- and HCO3-).

The anion gap in a healthy individual is around 15 mEq/l (range 8–18 mEq/l).

The alterations in this anion gap is mostly associated with acid-base disorders.

47
Summary

48
 Role of Hemoglobin to Maintains Acid-Aase Balance

Structure of Hemoglobin

Hb is a metalloprotein composed of four polypeptide

chains (globins).

It contains two α chains & two β chains.

Each globin is linked to a heme group that contains a

ferrous ion (Fe2+) held within a porphyrin ring.

49
 Cont’d…
Hemoglobin (Hb) molecule within RBCs:

carries O2 from the lungs to the tissues

transports CO2 from tissues back to the lungs

 This helps to maintain the acid-base balance.

Hb of erythrocytes is also important in the respiratory regulation of pH.

At the tissue level, Hb binds to H+ ions and helps to transport CO2 as HCO3-.

In the lungs, when hemoglobin combines with O2, H+ ions are removed from the

hemoglobin, and it will combine with HCO3- to form H2CO3.

50
 This H2CO3 latter dissociates to release CO2 to be exhaled.
  Carbonic anhydrase (CA) is the
enzyme that catalyzes the
conversion of CO2 to H2CO3.

 When CO2 diffuses as a dissolved

gas from the tissue capillaries, it
binds to the α-amino terminus of
the globulin chain, forming
Carbaminohemoglobin.

Generation of bicarbonate by the erythrocyte

51
 CO2 produced in tissues is converted to bicarbonate to be transported to the lungs.
52
 At the lungs, bicarbonate is converted back to CO2 and eliminated by the
lungs.
53
 Arterial Blood Gas Measurements (ABG Analysis)

In certain conditions like respiratory failure and/or acid-base disorders, blood
gas (CO2 & O2) measurements in the laboratory are significant.

To analyze the blood gases, a sample of arterial blood is needed.

The biochemical profile measured include pO2, pCO2, and pH.

Results of these three profiles allow the classification of acid-base

disturbance to one of the four etiological categories.

54
 Cont’d…

The Reference ranges of arterial blood gas analysis

Bicarbonate ion concentration is measured from the concentration of CO2 and

pH. How????
55
 Role of Pulmonary Surfactants in Respiration

Alveoli represent the basic units for gas exchange.

The surface area imposed by alveolar structures is basically linked to very high surface
tension at the air-water interface.
These surface tensions are opposed by pulmonary surfactants that
reduce surface tension.
 The term surfactant means a surface-active agent.

Surfactants are surface-acting agents formed by Type II alveolar cells.

56
 Cont’d...

Since surfactants have both a hydrophilic region (strongly attracted to water) and a
hydrophobic region (strongly repelled by water),
they are localized to the surface of an air-water interface (where the two
molecules meet).

The major components of pulmonary surfactant are lipids (90%) and proteins
(10%).

Dipalmitoylphosphatidylcholine (DPPC)/dipalmitoyl lecithin is the chief

phospholipid component. 57
 Cont’d...

Surfactants are synthesized from ER of type II alveolar cells (pneumocytes).

Type II cells are granular pneumocytes, round cells, and contain lamellar
inclusions.

These inclusions contain surfactant which is secreted from these cells by

exocytosis.

Type I alveolar cells are highly necessary for gas exchange.

58
Composition of Surfactants 59
 Cont’d...
Surfactants line the inner surface of the lung to prevent alveolar collapse at the end of
expiration.
 This is done by reducing the surface tension.

Surfactants have hydrophilic and hydrophobic regions.

The hydrophilic head faces towards the water and the hydrophobic tails face
towards the air.

Nonpolar tail Polar head

60
Cont’d...

61
 Biosynthesis of Surfactant Phospholipids

There are different substrates that are required for the biosynthesis of surfactants.
These are:

1. Fatty Acids: Type II cells can obtain FAs from the circulation or TAG of
lipoprotein.

2. Phosphatidic Acid

3. Choline: Primarily from the diet or from hydrolysis of phosphatidylcholine (PC).

4. Glycerol: Usually from glucose/glycogen.

62
 Steps of Biosynthesis of Surfactant

1. Activation of choline by choline kinase

2. Synthesis of Cytidindiphosphate (CDP)-choline by phosphocholine citidyl

transferase

3. Synthesis of PC by PC diacylglycerol (DAG) transferase

63
 Activation

Synthesis of
Cytidindiphosphat
e choline (CDP-C)

Synthesis of DPPC

64
Cont’d..

65
 Summary of DPPC Synthesis
Glucose Glycogen

NAD+ NADH
glycerol-3-phosphate DHAP
palmitoyl-CoA

CoASH
palmitoyl-G3P Choline
palmitoyl-CoA ATP

CoASH ADP
dipalmitoylphosphatidic acid phosphocholine
H2O CTP

Pi PPi
dipalmitoylglycerol Cytidindiphosphate
CMP choline (CDP-choline)
DPPC
66
 Functions of Surfactants

Surfactants increase pulmonary compliance (the ability of the lungs to

expand).
This is used to prevent atelectasis.

 Atelectasis is the complete or partial collapse of the entire lung or area.

Surfactants are also necessary for the regulation of the alveolar size.

They prevent fluid accumulation and keep airways dry.

67
 Hormones Affecting Surfactant Production

Thyroid Hormone: These hormones stimulate type II cells.

Cortisol Hormone: These hormones stimulate the production and maturation of

surfactants.
 In the fetus, shortly before birth, an increase in plasma cortisol levels
upregulates several enzymes that are important for the synthesis of
dipalmitoyl phosphatidylcholine (DPPC) including:
 FA synthase and phosphocholine transferase

68
 Clinical Significance of Pulmonary Surfactant

Hyaline-membrane disease (HMD)/Infant Respiratory distress syndrome (IRDS) is

the disorder:
Occurs primarily in premature infants.

Surfactant deficiency is the primary cause of HMD.

 If DPPC is absent, the alveolar radius becomes smaller.

With expirations, the wall tension rises and the alveoli collapse.

There is rapid or difficult of breathing immediately after birth, cyanosis,

tachycardia.
69
Cont’d…

70
 Evaluation of Fetal Lung Maturity
Lecithin to Sphingomyelin Ratio (L/S ratio) in amniotic fluid has been used for the
evaluation of fetal lung maturity

Before 34 weeks of gestational age, the concentrations of lecithin and sphingomyelin in

amniotic fluid are approximately equal.

After 34 weeks, there is a marked increase in lecithin, and the L/S ratio increases to
greater than 5 at term.

Therefore, when the fetal lung is evaluated, if:

L/S ≥2, there is adequate lung maturity.

L/S≤1, the infant will probably develop HMD.

71
 Cont’d...

There are different treatment options for Hyaline-membrane disease (HMD)/

Infant Respiratory distress syndrome (IRDS). These are:
Continues positive airway pressure (CPAP)

Glucocorticoids to the mother

Exogenous (artificial) surfactant replacement immediately after birth.

72
The End of The Topic

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