Lecture- MICROBIAL PHYSIOLOGYBY: Dr.

Maha Adel
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BACTERIAL STRUCTURE

The outer layer or cell envelope consists of two components, a rigid cell
wall and beneath it a cytoplasmic or plasma membrane. The cell envelope
encloses the protoplasm, comprising the cytoplasm, cytoplasmic inclusions
such as ribosomes and mesosomes, granules, vacuoles and the nuclear
body.

Cell wall
Beneath the external structures is the cell wall. It is very rigid & gives
shape to the cell. Its main function is to prevent the cell from expanding &
eventually bursting due to water uptake. Cell Wall constitutes a significant
portion of the dry weight of the cell and it is essential for bacterial growth
& division. The cell wall cannot be seen by direct light microscopy and does
not stain with simple stains. It may be demonstrated by microdissection,
reaction with specific antibodies, mechanical rupture of the cell,
differential staining procedures or by electron microscopy.
The surface of gram-negative cells is much more complex than that of
gram-positive cells.
The gram-positive cell surface has two major structures: the cell wall and
the cell membrane.
The cell wall of gram-positive cells is composed of multiple layers of
peptidoglycan, which is a linear polymer of alternating units of N-
acetylglucosamine (NAG) and N-acetylmuramic acid (NAM). A short
peptide chain is attached to muramic acid. A common feature in bacterial
cell walls is cross-bridging between the peptide chains. Embedded in it are
polyalcohol called Teichoic acids. Some are linked to Lipids & called
Lipoteichoic acid. Lipotechoic acid link peptidoglycan to cytoplasmic
membrane and the peptidoglycan gives rigidity.

The functions of Teichoic acid are

● gives negative charge
● major antigenic determinant
● transport ions
● anchoring
external permeability barrier ●

In a gram-positive organism such as Staphylococcus aureus, the cross-

bridging between adjacent peptides may be close to 100%. By contrast,

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the frequency of cross-bridging in Escherichia coli (a gram-negative
organism) may be as low as 30%.

Outer Membrane
Outer membrane is found only in Gram-negative bacteria, it functions as
an initial barrier to the environment and is composed of
lipopolysaccharide (LPS) and phospholipids.

Lipopolysaccharide (LPS)
The LPS present on the cell walls of Gram-negative bacteria account for
their endotoxic activity and antigen specificity.
A bacterium is referred as a protoplast when it is without cell wall. Cell
wall may be lost due to the action of lysozyme enzyme, which destroys
peptidoglycan. This cell is easily lysed and it is metabolically active but
unable to reproduce. A bacterium with a damaged cell wall is referred as
spheroplasts. It is caused by the action of toxic chemical or an antibiotic,
they show a variety of forms and they are able to change into their normal
form when the toxic agent is removed, i.e. when grown on a culture
media.
Cell Walls and Osmotic Protection
Microbes have several mechanisms for responding to changes
in osmotic pressure. This pressure arises when the concentration
of solutes inside the cell differs from that outside, and the
,responses work to equalize the solute concentrations. However
in certain situations, osmotic pressure can exceed the cell’s
ability to acclimate. In these cases, additional protection is provided
by the cell wall.
When cells are in hypotonic solutions Ones in which the solute
concentration is less than that in the cytoplasm—water diffuses into the
cell, causing it to swell. Without the peptidoglycan layer of the cell wall,
the pressure on the plasma membrane would become so great that the
membrane would be disrupted and the cell would burst—a process called
.lysis
Conversely, in hypertonic solutions, water flows out and the cytoplasm
shrivels up—a process called plasmolysis. The protective nature of
peptidoglycan is most clearly demonstrated when bacterial cells are
treated with lysozyme or penicillin. The enzyme lysozyme attacks
peptidoglycan by hydrolyzing the bond that connects N-acetylmuramic
acid with N-acetylglucosamine. Penicillin works by a different mechanism.
It inhibits the enzyme transpeptidase, which is responsible for making the

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cross-links between peptidoglycan chains. If bacteria are treated with
either of these substances while in a hypotonic solution, they
.lyse
However, if they are in an isotonic solution, they can survive and grow
normally. Treatment of typical Gram-positive bacteria with lysozyme or
penicillin results in the complete loss of the cell wall, and the cell becomes
a protoplast. When typical Gram-negative bacteria are exposed to
lysozyme or penicillin, the peptidoglycan sacculus is destroyed, but the
.outer membrane remains. These cells are called spheroplasts
Because they lack a complete cell wall, both protoplasts and
spheroplasts are osmotically sensitive. If they are transferred to
a hypotonic solution, they lyse due to uncontrolled water influx

Chlamydiae and Planctomycetes. Both have an outer membrane like

other Gram-negative bacteria, but there is no peptidoglycan layer between
the plasma membrane and outer membrane. Despite their lack of
peptidoglycan, chlamydia and planctomycetes maintain a characteristic
shape. This is not true of mycoplasmas which lack a cell wall altogether;
they tend to be pleomorphic. Mycoplasmas also are osmotically
sensitive. Despite this, they often can grow in dilute media or terrestrial
environments because their plasma membranes are more
resistant to osmotic pressure than those of walled bacteria. The
precise reason for this is not clear, although the presence of sterols
in the membranes of many species may provide added strength.

Cytoplasmic membrane is present immediately beneath the cell wall,

found in both Gram positive & negative bacteria and it is a thin layer lining
the inner surface of cell wall and separating it from cytoplasm. It acts as a
semipermeable membrane controlling the flow of metabolites to and from
the protoplasm.

The cytoplasmic membrane of both gram-positive and gram-negative cells

is a lipid bilayer composed of phospholipids, glycolipids, and a variety of
proteins. The proteins in the cytoplasmic membrane may extend through
its entire thickness. Some of these proteins provide structural support to
the membrane while others function in the transport of sugars, amino
acids, and other metabolites. The outer membrane of gram-negative cells
contains a relatively high content of lipopolysaccharides . These lipid-

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containing components represent one of the most important identifying
features of gram-negative cells: the O antigens, which are formed by the
external polysaccharide chains of the lipopolysaccharide. This lipid
containing component also displays endotoxin activity—that is, it is
responsible for the shock observed in severe infections caused by gram-
negative organisms. Bacterial cell surfaces also contain specific
carbohydrate or protein receptor sites for the attachment of
bacteriophages, which are viruses that infect bacteria. Once attached
to these receptor sites, the bacteriophage can initiate invasion of the cell
Gram-positive and gram-negative cells have somewhat different strategies
for transporting materials across the membrane and into the cell. The
cytoplasmic membrane of gram-positive organisms has immediate access
to media components. However, chemicals and nutrients must first
traverse the outer membrane of gram-negative organisms before
encountering the cytoplasmic membrane. Gram-negative cells have pores
formed by protein triplets in their outer membrane that will permit
passage of fairly large molecules into the periplasmic space. Subsequent
transport across the inner or cytoplasmic membrane is similar in both
.gram-positive and gram-negative cells

Cytoplasm
The cytoplasm is a Colloidal system containing a variety of organic and
inorganic solutes containing 80% Water and 20% Salts, Proteins. They are
rich in ribosomes, DNA & fluid. DNA is circular and haploid. They are highly
coiled with intermixed polyamines & support proteins. Plasmids are extra
circular DNA.

Ribosomes
They are the centers of protein synthesis. They are slightly smaller than
the ribosomes of eukaryotic cells. Ribosomes. The cytoplasm of all cells
has a fine granular appearance observed in many electron micrographs.
Tiny particles called ribosomes are responsible for this look.

Ribosomes contain approximately 65% RNA and 35% protein. The

ribosome orchestrates the polymerization of amino acids into proteins
(i.e., protein synthesis).

At higher magnification under the electron microscope the ribosome

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particles are spherical. In properly prepared specimens the ribosomes are
observed as collections or chains held together on a single messenger RNA
(mRNA) molecule and are referred to as polyribosomes or simply
polysomes.

The more or less spherical ribosome particle, when examined by sucrose

gradient sedimentation, has been found to have a svedberg coefficient of
70S. (A Svedberg unit denotes the rate of sedimentation of a
macromolecule in a centrifugal field and is related to the molecular size of
that macromolecule.) The prokaryotic ribosome may be separated into
two lower-molecular-weight components: one of 50S and another of 30S.
Only the complete 70S particle functions in polypeptide synthesis.
By comparison, the ribosomes of eukaryotic cells are associated with the
endoplasmic reticulum, are larger (80S), and are composed of 40S and 60S
subunits. The function of both 70S and 80S ribosomes in protein synthesis
is identical. Curiously, eukaryotic mitochondria characteristically display
70S ribosomes—not the 80s particles that you would expect—because
mitochondria probably evolved from endosymbiotic prokaryotic cells, a
hypothesis supported by extensive analyses comparing bacterial
and mitochondrial genomes.

Mesosomes
They are vesicular, convoluted tubules formed by invagination of plasma
membrane into the cytoplasm. They are principal sites of respiratory
enzymes and help with cell reproduction.

Inclusions:
are common in all cells. They are formed by the aggregation of substances
that may be either organic or inorganic. Inclusions can take the form of
granules, crystals, or globules; some are amorphous. Some inclusions lie
free in the cytoplasm. Other inclusions are enclosed by a shell that is
single-layered and may consist of proteins or of both proteins and
phospholipids. Some inclusions are surrounded by invaginations of the
plasma membrane.
The Inclusion bodies are aggregates of polymers produced when there is
excess of nutrients in the environment and they are the storage reserve
for granules, phosphates and other substances. Volutin granules are
polymetaphosphates which are reserves of energy and phosphate for cell
metabolism and they are also known as metachromatic granules.

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Lecture- MICROBIAL PHYSIOLOGYBY: Dr. Maha Adel
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Carbon is often stored as polyhydroxyalkonate (PHA) granules. Several
types of PHA granules have been identified, but the most common contain
poly-b-hydroxybutyrate (PHB).

Nucleus

The Nucleus is not distinct and has no nuclear membrane or nucleolus and
the genetic material consist of DNA. The cytoplasmic carriers of genetic
information are termed plasmids or episomes

Capsule

Capsule is the outer most layer of the bacteria (extra cellular). It is a

condensed well defined layer closely surrounding the cell. They are usually
polysaccharide and if polysaccharide envelops the whole bacterium it is
capsule and their production depends on growth conditions. They are
secreted by the cell into the external environment and are highly
impermeable. When it forms a loose mesh work of fibrils extending
outward from the cell they are described as glycocalyx and when masses
of polymer that formed appear to be totally detached from the cell and if
.the cells are seen entrapped in it are described as slime layer

Capsules are composed of either polysaccharides (high molecular- weight

polymers of carbohydrates) or polymers of amino acids called
polypeptides (often formed from the D- rather than the L-isomer of an
amino acid). The capsule of Streptococcus pneumoniae type III is
composed of glucose and glucuronic acid in alternating β-1, 3- and β-1, 4-
linkages: This capsular polysaccharide, sometimes referred to as
pneumococcal polysaccharide, is responsible for the virulence of the
pneumococcus. Bacillus anthracis, the anthrax, bacillus, produces a
polypeptide capsule composed of D-glutamic acid subunits, which
is a virulence factor for this organism.
The Capsule protects against complement and is antiphagocytic. The Slime
layer & glycocalyx helps in adherence of bacteria either to themselves
forming colonial masses or to surfaces in their environment and they
resists phagocytosis and desiccation of bacteria
Flagella

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Lecture- MICROBIAL PHYSIOLOGYBY: Dr. Maha Adel
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Flagella are long hair like helical filaments extending from cytoplasmic
membrane to exterior of the cell. Flagellin is highly antigenic and functions
in cell motility. The location of the flagella depends on bacterial species as
polar situated at one or both ends which swims in back and forth fashion
.and lateral at along the side
The parts of flagella are the filament, hook and the basal body. Filament is
external to cell wall and is connected to the hook at cell surface, the hook
& basal body are embedded in the cell envelope. Hook & filament is
composed of protein subunits called as flagellin. Flagellin is synthesized
.within the cell and passes through the hollow centre of flagella

The hook and basal body are quite different from the filament
Slightly wider than the filament, the hook is made of different protein
subunits. The basal body is the most complex part of a flagellum. The basal
bodies of E. coli and most other typical Gram-negative bacteria have four
rings: L, P, MS, and C which are connected to a central rod. The L, P, and
MS rings are embedded in the cell envelope, and the C ring is on the
cytoplasmic side of the MS ring. Typical Gram-positive bacteria have only
two rings: an inner ring connected to the plasma membrane and an outer
one probably attached to the peptidoglycan. The synthesis of bacterial
flagella is complex and involves at least 20 to 30 genes. The arrangement
of flagella may be described as
Monotrichous – single flagella on one side )i(

Lophotrichous – tuft of flagella on one side )ii(

Amphitrichous – single or tuft on both sides )iii(

Peritrichous – surrounded by lateral flagella )iv(

Flagella Type Example

Monotrichous: Vibrio cholerae

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Lecture- MICROBIAL PHYSIOLOGYBY: Dr. Maha Adel
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Lophotrichous: Bartonella , bacillifornis
Amphitrichous: Spirillum serpens
Peritrichous: Escherichia coli

Various types of mobility is observed because of the presence of the

flagella as Serpentine motility is seen with Salmonella, Darting motility
with Vibrio and Tumbling motility with Listeria monocytogenes

Pili / Fimbriae

Hair-like proteinaceous structures that extend from the cell membrane to

external environment are pili which are otherwise known as fimbriae. They
are thinner, shorter and more numerous than flagella and they do not
function in motility. The fimbriae is composed of a subunit called pilin
There are two types pili namely Non-sex pili (Common pili) eg. fimbriae or
type IV and the sex pili. The fimbriae are antigenic and mediate their
.adhesion which inhibits phagocytosis. The sex pili help in conjugation
Spore
Some bacteria have the ability to form highly resistant resting stage called
spores, which helps them to overcome adverse environmental conditions
that are unfavorable for vegetative growth of cell. They are not a
reproductive form and not a storage granule. These spores are resistant to
bactericidal agents and adverse physical conditions. Each spore can give
rise to only one endospore which play a role in heat resistance.
Spores consists of three layers namely core, cortex and spore coat
The ability of the spore to survive heat, radiation, and
damaging chemicals requires that its enzymes and DNA be
protected. The various layers of the spore contribute to this
resistance in several ways:
The spore coat protects the spore from chemicals and
various lytic enzymes such as lysozyme.
The inner membrane is extremely impermeable to various
chemicals, including those that cause DNA damage.
The core has very low water content, high amounts of
dipicolinic acid complexed with calcium ions (Ca- DPA), and
a slightly lower pH, all of which contribute to the spore’s
resistance to harsh conditions. Evidence exists that the

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water content of the core is low enough to prevent rotation
of enzymes and other proteins present.