BASIC

BACTERIOLOGY

R.S. Maghembe and D G K. Ghati

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 Brief Introduction
• All living cells (despite their complexity
and variety) can be grouped into two;
• Prokaryotic
• Eukaryotic.
• We will describe structure of the cell from
the inside to the outside

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THREE-DOMAIN SYSTEM

Carl Woese, George E. Fox et al. in 1990.

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 Typical bacteria
•Have normal peptidoglycan
•May be normal flora or may be
pathogenic in humans.
•Do not have a sexual growth cycle;
however, some can produce asexual
spores.
Mycoplasmas
•Are the smallest and simplest of the
bacteria that are self-replicating.
lack a cell wall.
•Are the only prokaryotes that contain
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sterols. RSM & DGK 4
 Rickettsia spp
•Are obligate intracellular bacteria that are
incapable of self-replication.
•Depend on the host cell for adenosine
triphosphate (ATP) production
•Cause spotted fever: Transmitted by ticks
Chlamydia spp
•Are obligate intracellular pathogens with
a complex growth cycle involving
intracellular and extracellular forms.
•Depend on the host cell for ATP
production
•Cause urinogenital infections, e.g. PID
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Eukaryotic cell structure

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 Eukaryotic Cell Structure
Nucleus
– Is bound by membrane continuous with ER
– Nuclear membrane exhibits selective permeability
due to pores permitting exchange of molecules
between the nucleus and cytoplasm
– Chromosomes contain linear DNA macromolecules
arranged in double helix, associated with proteins
(histones) that bind DNA by ionic interactions
– Chromosomes are visible with light microscope
during cell division (when DNA is in a highly
condensed form)

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 Eukaryotic Cell Structure
Cytoplasm
• Characterized by the following features:
– Endoplasmic reticulum
• Is a network of membrane-bound channels
• The membranes are coated with ribosomes in
some regions
• Ribosomes provide site for protein synthesis
• Synthesized proteins pass through the membrane
into the channels of endoplasmic reticulum and
transported to other parts of the cell

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 Eukaryotic Cell Structure
Cytoplasm cont…
• Golgi Apparatus
– This is related to the ER
– It pinches off vesicles that can fuse with the cell
membrane and release the enclosed biomolecules
into the surrounding medium

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 Eukaryotic Cell Structure
• Cytoplasm cont…
– Mitochondria
• Is included in Plastid
• Contain the respiratory electron transport system
• The plastid also contains chloroplasts (in
photosynthetic organisms)
• Plastids contain own DNA which codes for some
(but not all) of their constituent proteins and
tRNAs

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 Eukaryotic Cell Structure
• Cytoplasm cont…
Cytoskeleton:
 Composed of three different types of
filaments
a)Microtubules
• Cytoplasm includes array of microtubules
which play important role in:
– Cytoplasmic membrane function and cell
shape,
– Formation of mitotic spindles and
– Motility e.g. flagella

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 Eukaryotic Cell Structure
• Cytoplasm cont…
Cytoskeleton:
b. Microfilaments
• Also known as actin
• Provide structural support and mortility
• Composed of polymerized α- and β-
tubulin
• Membrane-microfilament binding proteins
connect them to the membrane

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 Eukaryotic Cell Structure
• Cytoplasm cont…
Cytoskeleton:
c. Intermediate filaments

• Support for Nuclear membrane

– Lamins

• Cell adhesion
– Desmin, keratin, vimentin.
• Motility
– MSP

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 Surface layers

• The cytoplasm is enclosed within plasma membrane

composed of proteins and phospholipids
• Most animal cells do not have other surface layers but
plant cells have an outer cellulose cell wall
• However, many eukaryotic microorganisms have an
outer cell wall in additional to the plasma membrane
– This cell wall may be composed of a polysaccharide
such as cellulose, or chitin (fungi) or may be may
be inorganic (silica wall of diatoms)

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 Motility organelles
• Many eukaryotic MOs have flagella or cilia which
move with a wave-like motion to propel the cell
• Eukaryotic flagellae emanate from polar region of the
cell whereas cilia surround the cell
• Both flagella and cilia of eukaryotic cells have the same
basic structure and biochemical composition
• Both consist of a series of microtubules (hollow
proteins cylinders) composed of a protein called tubulin
surrounded by a membrane
• Arrangement of microtubules is called the “9 + 2
system” because of the 9 peripheral pairs of
microtubules surrounding 2 single central microtubules
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 • Left: A zoospore of
the fungus, Allomyces,
with a single flagellum.
• Right: a partially
disintegrated flagellum
of Allomyces showing
2 inner fibrils and 9
outer fibrils

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 Eukaryotic Cell Structure

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 Two functions of ribosomes
1. Decoding the genetic code in mRNA

2. Catalyzing the formation of peptide bonds

between amino acids resulting in a
polypeptide chain

–In essence, a ribosome is an enzyme,

–A Polypeptide Polymerase

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 Functional significance of the ribosomal structure

• Ribosomes have two subunits, large and small

– Prokaryotic: 70S, comprising of 50S & 30S
– Eukaryotic: 80S, comprising of 60S & 40S

– The Peptidyl transferase center and the catalytic

site are in the large s/unit (60S / 50S)

– The small subunit (40S / 30S) serves as the

assembly guide for factors needed in protein
synthesis: decoding the mRNA also occurs on
the small subunit RSM & DGK
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 Components of Ribosomes

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 PROKARYOTIC CELL
STRUCTURE
• Prokaryotic cell is simpler than eukaryotic cell at every level
except the cell envelop (which is more complex)
• Consist of vast heterogeneous groups of unicellular organisms
(bacteria)
• Bacteria are single-celled organisms:-
– Contain no membrane-bound nucleus
• Termed prokaryotes = pre nucleus (Pro = pre and karyote
= nucleus)
– Average size is 0.5 - 2.0 µm in diameter
• Compare with RBC size 7.5 µm in diameter
– Cytoplasm is surrounded by lipid membrane surrounded by a
rigid cell wall
– DNA is bundled in a region called nucleoid
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• Several species of bacteria are
differentiated by such factors as:
• Morphology
• Chemical composition\staining
properties
• Nutrition requirements
• Biochemical activities

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 Nuclei of Bacillus cereus
dispersed within cytoplasm
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 The Bacterial Cell

Flagella

Ribosomes

Capsule

Cell wall
Plasma
membrane

Nucleoid region (DNA)

Pili

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 The Bacterial Cell
Nucleoid DNA

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 Bacterial Morphology
• Morphology is classification by shape and is often
expressed in the binomial naming system
– Example
• Bacillus subtillis, Bacillus means ‘rod’
• Streptococcus pyogenes, Streptococcus
means a chain of spheres
• Helicobacter pylori, helix shaped bacterium
– Many names of bacteria do not refer to their
shapes such as:
• E. coli, Pseudomonas aeruginosa
• or Burkholderia cepacia.
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 Bacterial Morphology

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 Examples of Morphology
Various Bacterial shapes:
• Cocci
• Spherical
• In chain = Streptoccus
• In cluster = Staphylococcus
• Cocci are grouped into:
• Diplococci
• Streptococci
• Tetrads - divided into planes
• Sardine- three regular planes-box-like
• Staphylococci - random planes
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• Bacillus
• Rod or cylinder shaped
– Bacilli can only be divided
across their axis
– Bacilli can be grouped into:
• Diplobacilli
• Streptobacilli (chain)
• Coccobacilli
– Short round rod, oval and
look like cocci

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 Other shapes
• Vibrio
• Curved rod
• Spirillum
• Spiral shaped
• Spirochete
• Helical shape
• Square
• Flat cells genus Arcula
• Star shaped
• Genus Stella
• Pleomorphic
• Several shapes; bacterial shape
are determined by environment
e.g. Rhizobium and
Corynebacterium

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 Bacterial Nomenclature
• Binomial naming system
– ‘Two word’ naming system
• First word is genus name
– Always capitalized
• E.g. Escherichia
• Second word is species name (specific epithet)
– Not capitalized
• E.g. coli
• When writing full name genus usually abbreviated
– E. coli
• Full name always italicized or underlined

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 Bacterial Anatomy
Cytoplasm
• It is an internal matrix of the cell contained inside the
plasma membrane
• Is the thick, semitransparent, elastic fluid that fills
the cell where most cellular reactions and activities
take place
• Is made up of 80% water, and proteins (enzymes),
carbohydrates, lipids, inorganic ions and many low-
molecular-weight compounds
• The cytoplasm has two distinct areas when seen in
the electron microscope:-
1. An amorphous matrix containing ribosomes, nutrient
granules, metabolites and plasmids
2. An inner nucleoid region composed of DNA
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 Nucleoid
• Is an area of cytoplasm where DNA is located
• It is equivalent to the nucleus in eukaryotic
cells
• Can be seen with the light microscope when
stained
• It is Feulgen-positive, indicating the presence
of DNA
• Study by electron microscope reveals that
prokaryotic cell do not have a nuclear
membrane and a mitotic apparatus
• The nuclear region is filled with DNA fibrils
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 Nucleoid cont...

• DNA is a single-stranded, haploid chromosome,

circular molecule (Mwt about 2-3 x 10 9) and
contain approx. 2000 genes, with no histones
Note:
– Human DNA has approximately 100,000 genes
– Some prokaryotes such as Borrelia burgdorferi,
(agent of Lyme disease) and several
Streptomyces species, have a linear chromosome

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 Plastids
• Prokaryotic cells do not have mitochondria or
chloroplast
– The electron transport enzymes are localized
in the cytoplasmic membrane
– The photosynthetic pigments (Carotenoids,
Bacterial chlorophyll) of photosynthetic
bacteria are located in specialized membrane
arrangements that may appear as spherical
vescicles or as flattened sheet-like layers
underlying the cell membrane

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 Plastids cont...

• In some Cyanobacteria (blue-green

algae), the photosynthetic membrane
forms multilayered structures called
Thylakoids
• The major accessory pigments used for
light harvesting are Phycobilins found
on outer surface of the thylakoid
membrane
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 Granules
• Cytoplasm contains several different types of granules
functioning as storage area for nutrients
• Granules stain characteristically with certain dyes
• Bacteria store reserve materials in form of insoluble
granules deposited as osmotically inert neutral
polymers
• When source of nitrogen, sulfur, or phosphorus is
limited or when pH is low, excess CO2 in the medium
is converted by some bacteria to the polymer “Poly-β-
hydroxybutric acid and by other bacteria to various
polymers of glucose such as starch and glycogen

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 Granules cont...
• Such granules serve as carbon source when protein and nucleic acid
synthesis is resumed
• Additionally, certain photosynthetic bacteria oxidize sulfide from H2S
to produce intracellular granules of sulfur elements
• Many bacteria accumulate granules of polyphosphate (high energy
stored in form of polymerized metaphosphate) which are reserves of
inorganic phosphate, used in ATP synthesis
• These granules are known as Volutin Granules or Metachromatic
Granules because they stain red with a blue dye such as Methylene
blue.
• Volutin granules are characteristic features of Corynebacteria e.g.
Corynebacterium diphtheriae, which causes diphtheria in human

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 METACHROMATIC GRANULES

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 Ribosomes

• Translate mRNA into a sequence of amino acids

(proteins)
• Bacterial ribosomes are thus, the site for protein
synthesis, just like in eukaryotic cells
• Prokaryotic ribosomes are smaller than eukaryotic
ribosomes and have a different chemical composition
• Bacterial (prokaryotic) ribosomes are 70S in size
with 50S and 30S subunits whereas eukaryotic
ribosomes are 80S with 60S and 40S subunits

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 Medical & Pharmaceutical Importance of
Ribosomes
• The difference in ribosomal RNAs and
protein form the basis for “Selective
Action” of several antibiotics inhibiting
bacterial protein synthesis
• Examples of such antibiotics include:
– Chloramphenicol and macrolides
which bind to 50S subunit (only
found in bacteria)
– Aminoglycosides which binds to
30S subunit
• Details on Selective Action or Toxicity
of drugs are covered in
PHARMACOLOGY lectures

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 Plasmids
• They are extrachromosomal, double-stranded, circular
DNA molecules capable of replicating independently of
the bacterial chromosome
– Can be integrated into the bacterial chromosome
• Are typically thousands of times smaller than nucleoid
DNA
• Are very important as may carry genes for antibiotic
drug resistance
• Occur in both Gram +ve and Gram –ve bacteria
• Several different types of plasmids can exist in one cell

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 Types of plasmids
• Transmissible Plasmids
– Transferrable from one cell to another by conjugation
– Are large as they contain about a dozen genes
responsible for synthesis of the sex pilus and
enzymes required for transfer
– Usually present in a few (one to three) copies per cell
• Nontransmissible Plasmids
– Are small
– Do not contain the transfer genes
– Frequently present in many (10 – 60) copies per cell
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 Medical/pharmaceutical Importance of
Plasmids

• Plasmids carry genes for the following functions and structures:-

1. Antibiotic resistance, mediated by a variety of enzymes
2. Resistance to heavy metals such as Mercury (active component
of some antiseptics such as Merthiolate and Mercurochrome)
and Silver, mediated by a reductase enzyme
3. Resistance to UV-light, mediated by DNA repair enzymes
4. Pili (Fimbriae), mediate attachment and adherence of bacteria
to epithelial cells
5. Exotoxins – including several enterotoxins

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 Other products encoded by
Bacteriocin
plasmids
– Toxins or enzymes produced by certain bacteria and
are lethal (i.e. kill) to other bacteria
Several antibiotics
– Produced by Streptomyces e.g. Streptomycin from
Streptomyces griseus
• Variety of degradative enzymes
– Produced by Pseudomonas, capable of cleaning up
environmental hazards such as oil spills and toxic chemical
wastes
• Tumors caused by Agabacterium in plants
• Nitrogen fixing enzymes in Rhizobium in the root
nodules of legumes
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 Transposons
• Are pieces/sequences of DNA that move rapidly from
one site to another, within or between DNAs of
bacteria, plasmids and bacteriophages.
• Are also known as “jumping genes”
• The movement is known as transposition
• They can perform the following:
– Code for drug resistance enzymes, toxins or
metabolic enzymes
– Cause mutations in genes into which they insert or
alter expression of nearby genes
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 Note
• Unlike plasmids or bacterial viruses,
transposons are incapable of independent
replication
• They replicate as part of recipient DNA
• More than one transposon can be located in
DNA e.g. a plasmid can contain several
transposons carrying drug resistance genes

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 The Cytoplasmic Membrane
• It is the barrier between the internal cell fluid
(cytoplasm) and exterior environment.
• It is selectively permeable only allowing in selected
compounds
Structure
– Bacterial (prokaryotic) cytoplasmic membrane is
also known as the cell membrane
– Is visible in electron micrographs of thin section
– It is a “typical UNIT MEMBRANE”, composed of
phospholipids and proteins
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 Membrane organization

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 Membrane organization

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 Membrane organization

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Cytoplasmic Membrane cont...

• The cytoplasmic membrane of prokaryotic cells

differ from that of eukaryotic cells in that;
– Prokaryotic membrane do not contain sterols
(except in Mycoplasmas) whereas that of
eukaryotic cells contain sterols
• Mycoplasmas are the only prokaryotes
containing sterols e.g. Cholesterol in their
membranes when growing in sterol-containing
media
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 Cytoplasmic Membrane
cont...
• Mesosomes
– Are specialized structures formed by convoluted
invaginations of cytoplasmic membrane
– Are important during cell division
– Are divided into two types:-
• Septal mesosomes – form cross-walls during cell
division
• Lateral mesosomes
– Generally, mesosomes function as the origin of
transverse septum that divides the cell into half and as
the binding site of the DNA that become the genetic
material of each daughter cell (ref cell division process
for details)
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 Roles of Mesosomes
• May start formation of transverse septum
and attach DNA to the plasma membrane
during binary fission
• Help to separate DNA into each daughter
cell
• Serve as generator of ATP and secrete
enzymes from the cell
• Enhance concentration of nutrients due to
the increase of surface area of plasma
membranes
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 Functions of Cytoplasmic
• Major functions:-
Membrane
1. Selective Permeability and Transport of Solute
– By providing hydrophobic barrier impermeable to most
hydrophilic molecules
– Several mechanisms (transport systems) enable transport
of nutrients into and waste products out of the cell
– These transport systems work against conc. gradient to
increase conc. of nutrients inside the cell (this function
requires energy)
– There are four general transport mechanisms involved in
membrane transport namely:-
– Facilitated diffusion
– Binding protein-dependent transport
– Chemiosmotic-driven transport and
– Group translocation

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 Functions cont...
• Facilitated diffusion
– Is the only transport mechanism that doesn’t
require energy
– It is a passive diffusion of substrates against a
concentration gradient
– The substrate NEVER achieves an internal
concentration greater than that outside the cell
– Glycerol is one of few compounds that enter
prokaryotic cells by facilitated diffusion

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 Functions cont...
• Binding Protein-dependent Transport
– Seen in Gram negative bacteria in which transport
of many nutrients is facilitated by specific binding
proteins located in the periplasmic space
– The proteins transfer bound substrate to compatible
membrane-bound transport protein complex
– The transport process is energized by ATP or other
high-energy phosphate compounds such as
Acetylphosphate

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 Functions cont...
• Chemiosmotic-driven Transport
– Moves molecules across the cytoplasmic
membrane at expense of previously
established ion gradient such as proton-motive
or sodium-motive force
– Three basic types of Chemiosmotic-driven
transport exist:-
1.Uniport – Catalyses transport of a substrate
independent of any coupled ion

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 Functions cont...
2. Symport –Catalyses co-transport of two
dissimilar species usually a solute and a
positively charged ion (e.g. H+) in the same
direction.
 i.e. two substrates are simultaneously
transported in the same direction by a single
carrier
 H+ gradient can permit symport of an
oppositely charged ion (e.g. glycine) or a
neutral molecule (e.g. galactose)

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 Functions cont...
3. Antiport – Catalyses simultaneous transport of
two like-charged compounds in opposite
direction by a common carrier (e.g. H+, Na+).
Note:
 Approx. 40% of substrates transported by E. coli
use this mechanism
 Above described transport mechanisms are
collectively referred to as “True Transport
Systems” because a solute ( molecule) is moved
across the membrane without change in structure

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 Transport Mechanisms

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 Group Translocation
• Is a process involving a net uptake of sugars
such as glucose and mannose
• The substrate is phosphorylated during the
transport process
• The process allows bacteria to utilize their
energy resources efficiently by coupling
transport with metabolism

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The Process
• A membrane carrier protein is first
phosphorylated in the cytoplasm at the
expense of phosphoenolpyruvate
• Then, the carrier binds the free sugar at
the exterior membrane face and transports
it into the cytoplasm and releases it as
sugar-phosphate complex

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 Major functions cont...
2. Electron Transport and Oxidative
Phosphorylation
• Cytochromes, other enzymes and components of
respiratory chain are located in the cytoplasmic
membrane
• In bacteria, cytoplasmic membrane is a functional
analog of the mitochondrial membrane
Note:
This relationship is thought to support the theory that
mitochondria have evolved from symbiotic bacteria

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 Major functions cont...
3. Excretion of Hydrolytic exoenzymes
• For degradation of polymers e.g. proteins,
polysaccharides, lipids etc. to subunits small enough
to penetrate cytoplasmic membrane
• Higher animals secrete enzymes into lumen of
digestive tract (e.g. alimentary canal in man)
• Bacteria secrete the enzymes directly into the
external medium ( in case of Gram positive cells) or
into the periplasmic space (space between outer
membrane of the cell wall and peptidoglycan layer)
in case of Gram negative bacterial cells
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 Major functions cont...
Note:
• Some Gram negative bacteria e.g. Pseudomonas,
Erwinia, and Serratia secrete large amount of
proteases, amylases and pectinases into the
extracellular environment
• These proteins (enzymes) are synthesized in the
cytoplasmic ribosomes
• Many pathogenic bacteria secrete enzymes (e.g.
IgA1 protease) and toxins (e.g. cholera toxin)

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 Major functions cont...
4. Biosynthetic Function
Cytoplasmic membrane is the site for carrier
lipids on which the subunits of the cell wall are
assembled and of the enzymes of the cell wall
biosynthesis. i.e. is the site for synthesis of cell
wall precursors
Enzymes of phospholipids synthesis are also
localized in the cytoplasmic membrane
Some proteins of the DNA replicating complex
are present at discrete sites in the membrane

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 Major functions cont...
5. Chemotactic Systems
– Attractants and repellents bind to specific
receptors in bacterial membrane
– For example; at least 20 different
chemoreceptors are found in E. coli
membrane
•Some of these chemoreceptors function
as first line step in the transport process

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 Antimicrobial Agents Affecting Cell
Membrane
Detergents
– Contain lipophilic and hydrophilic groups,
– Disrupt cytoplasmic membrane and kill the cell
Antibiotics (Some)
• Polymyxins, (Polymyxin B, Colistin)
– Are produced by the Gram-positive bacterium,
Bacillus polymyxa
– Consist of a cyclic peptide and a long hydrophobic
tail (detergent-like) that selectively damage
membranes containing phosphatidylethanolamine
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 Antibiotics cont...
– Are selectively toxic to Gram-negative bacteria
due to their specificity for the lipopolysaccharide
molecule within many Gram-negative outer
membranes.
• Daptomycin is an antibiotic class known as cyclic
lipopeptides, bind to bacterial membrane and causes
rapid membrane depolarization
• Other antibiotics specifically interfere with
biosynthetic function of the cytoplasmic membrane
– E.g. Nalidixic acid and Novobiocin inhibit DNA
synthesis
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 Antimicrobial Agents Affecting Cell
Membrane cont...
Ionophores
• Are membrane-active agents that permit rapid diffusion of specific
cations through the membrane
– E.g. Valinomycin specifically mediate passage of potassium ions
• Some ionophores act by forming hydrophilic pores in the membrane
• Others act as lipid soluble ion carriers
• Ionophores can kill cells by discharging membrane potential which
is essential for oxidative phosphorylation and other membrane
mediated processes
Note:
• Ionophores are NOT selective for bacteria, but act on membranes of
ALL cells

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 The Cell Wall
• Is a rigid, multilayered structure surrounding the cell
membrane that gives bacteria their shape.
• It is the outermost component of all bacteria (Except
Mycoplama species which are bound by a cell
membrane, not a cell wall)
• Is stronger than cell membrane, helps hold the cell
together and maintain structural integrity
• Differ in thickness depending on type of bacteria
• Some bacteria have surface features external to the
cell wall e.g.Capsule, Flagella, and Pili

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 The Cell Wall
cont...
• Is composed of inner layer of peptidoglycan and an
outer membrane
• The outer membrane varies in thickness and chemical
composition depending on bacterial type
• Peptidoglycan provides structural support and
maintains characteristic shape of the cell
• Peptidoglycan is unique to bacteria
Importance of peptidoglycan
• Provides high tensile strength to cell wall and prevent
bursting of the cell due to internal osmotic pressure
(internal osmotic pressure is due to solute
concentration via active transport)
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 Cell Wall of Gram +ve and Gram -ve Bacteria
• Note:
– Bacteria are classified as Gram –positive and Gram –
negative according to their response to the Gram staining
procedure
A: Peptidoglycan Layer
• Peptidoglycan is a complex polymer of amino acids and sugars
• The sugars are:
• N-acetylglucosamine (NAG) and
• N-acetylmuramic acid (NAM)
• The amino acids are of the “D form” and NOT “L form”
because the “D form” are difficult to break
• The D form amino acids cross link the NAG & NAM to form
an interwoven network surrounding the entire cell
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 Biosynthesis of
•
Peptidoglycan
Each peptidoglycan layer is a single, very big molecule
• In Gram positive bacteria there are up to 40 sheets of
peptidoglycan layers making up to 50% of cell wall
material
• Peptidoglycan is only 5-10% of the cell wall material
• Thus, cell wall of Gram-positive bacteria is mainly
peptidoglycan
• The layers are cross-linked by transpeptidation process
• In Staphyloccocus aureus transpeptidation occurs
between amino group on the end of pentaglycin cross-
link and the terminal carboxyl group of D-alanine on
the tetrapeptide side chains by action transpeptidase
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 Peptidoglycan layer cont...
• The peptidoglycan layer is composed of three parts:
1. A backbone composed of alternating N-
acetylglucosamine and N-acetylmuramic acid
2. A set of identical tetrapeptide side chains
attached to N-acetylmuramic acid and,
3. A set of identical peptide cross-bridges

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 Note:
• The carbohydrate backbone is the same in ALL
bacterial species
• The tetrapeptide side chains and the peptide
cross-bridges vary from species to species
• In many Gram negative cell walls the cross-
bridge consists of a direct peptide linkage
between Diaminopimelic acid (DAP) amino
group of one side chain and the carboxyl group
of the terminal D-alanine of a second chain

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• Tetrapeptide side chains of all species have following
features in common:
– Most have L-Alanine at position 1 (attached to NAM)
– Most have D-glutamate or substituted D-glutamate at
position 2
– D-Alanine at position 4
– Position 3 is the most variable position
• Most Gram negatives have diaminopimelic acid at
position 3 to which is linked the lipoprotein cell
component
• Gram positive bacteria may have DAP, L-lysin or
any of several other L-amino acids at position 3
11/11/24 RSM & DGK 82
 Peptidoglycan lattice formed by cross-linking. Bridges composed of
pentaglycin peptide chains connect the α-carboxyl of the terminal D-
alanine residue of one chain with the amino group of the L-lysine residue
of the next chain. Nature of cross-linking bridges varies among different
species
11/11/24 RSM & DGK 83
11/11/24 RSM & DGK 84
11/11/24 RSM & DGK 85
11/11/24 RSM & DGK 86
11/11/24 RSM & DGK 87
 Pharmaceutical Importance of
Peptidoglycan
• Provides side of action for:
– Penicillins and cephalosporins
• Act by inhibiting transpeptidase enzymes
which catalyses final cross-linking step in
synthesis of peptidoglycan
– Vancomycin
• Inhibit cell wall synthesis by interfering with
second stage of synthesis of peptidoglycan
(site earlier than that of penicillin)

11/11/24 RSM & DGK 88

 Special Components of Gram +ve
Cell Walls

1.Teichoic and Teichuronic Acids

• Are water soluble polymers containing ribitol or
glycerol residues joined through phosphodiester
linkages and carrying one or more amino acid or sugar
substituent

11/11/24 RSM & DGK 89

 Teichoic Acids
• Are polysaccharides of glycerol phosphate or ribitol
phosphate linked via phosphodiester bonds.
• Are located in the outer layer of Gram positive cell wall and
extend from it
• Are NOT found in Gram negative cells
• There are two types of teichoic acids
1.Wall teichoic acid
»Covalently linked to peptidoglycan
2.Membrane teichoic acid
»Covalently linked to membrane glycolipid and
concentrate in mesosomes
Note:
– Some Gram positive species lack wall teichoic acid but all
appear to contain membrane teichoic acid

11/11/24 RSM & DGK 90

 Teichoic Acid molecule

11/11/24 RSM & DGK 91

Teichoic Acids cont...
• Teichoic acid can be either covalently bonded
to N-acetylmuramic acid of the peptidoglycan
layer, linked to the plasma membrane lipids
found in the cytoplasmic membrane, or linked
to a terminal D-alanine in the tetrapeptide
cross-links between molecules of N-
acetylmuramic acid.
• Combined units of teichoic acids and lipids are
referred to as lipoteichoic acids
11/11/24 RSM & DGK 92
11/11/24 RSM & DGK 93
 Teichoic Acids cont...
• Teichoic acids are antigenic and induce antibodies that are
species specific
• Examples:
– In S. aureus, TAs mediate adherence to host mucosal cells
– In Pneumococcus e.g. Streptococcus pneumoneae TAs bear
an antigenic determinant called Forssman antigen
– In Streptococcus pyogenes, lipoteichoic acid is associated
with a protein called M protein, protruding from cell
membrane through peptidoglycan layer to form microfibrils
that facilitate attachment of S. pyogenes to animal cells

11/11/24 RSM & DGK 94

 Teichuronic Acids
• Similar to teichoic acids but contain sugar
acids as N-acetylmannosuronic or D-
glucosuronic acid instead of phosphoric
acids
• Teichuronic acids are synthesized in place of
teichoic acid when phosphate is limiting

11/11/24 RSM & DGK 95

Functions of Teichoic Acids
• Main function is to provide rigidity to cell-wall
• Bind magnesium ions and may play a role in supply
of Mg 2+ to the cell
• Assist in regulation of cell growth by limiting ability
of autolysins to break the β-1,4 bond between the N-
acetylglucosamine and the N-acetylmuramic acid.
• Serve as an attachment site for some parasites.
• Destruction of bacteria and release of Teichoic acid
into bloodstream may cause fever, blood vessel
dilation and possibly shock and subsequent death.

11/11/24 RSM & DGK 96

 2. Polysaccharides
 When Gram positive cell walls of certain
species are hydrolyzed, neutral sugars such
as mannose, arabinose, galactose, rhamnose
and glucosamine; and acid sugars such as
glucuronic acids and mannuronic acids are
obtained
 These sugars are proposed to exist as
subunits of polysaccharides in the cell wall

11/11/24 RSM & DGK 97

 Special Components of Gram
negative Cell Calls

• Gram negative cell walls contain three

components lying outside peptidoglycan layer:-
I. Lipoprotein
Cross-links the outer membrane and
peptidoglycan layer
Functions to stabilize the outer membrane
and anchor it to the peptidoglycan layer

11/11/24 RSM & DGK 98

 II. Outer Membrane
• Bacterial outer membrane is a bilayer structure found in Gram-
negative bacteria
• Is linked to the cell's peptidoglycan by Braun's lipoprotein
• Its composition is distinct from that of the cytoplasmic membrane
– Its inner “leaflet” has a composition identical to that of the
cytoplasmic membrane
– But phospholipids of outer leaflet are replaced by a complex
lipopolysaccharide (LPS) whose lipid portion acts as an endotoxins
– Thus, leaflets of outer membrane are asymmetrical and properties of
this bilayer differ from that of symmetrical cytoplasmic membrane

11/11/24 RSM & DGK 99

 100
11/11/24 RSM & DGK 100
 Bacterial Outer Membrane

101
11/11/24 RSM & DGK 101
 Outer Membrane cont...
• Outer membrane excludes hydrophobic molecules and
protect the cell (in case of enteric bacteria) from bile
salts
• Due to its lipid nature, outer membrane would be
expected to exclude hydrophilic molecules BUT it
permits passive diffusion of low molecules hydrophilic
compounds such as sugars, amino acids, & some ions
through special channels made of protein molecules
called porins
What about passage of antibacterials?
• Large antibiotic molecules penetrate the outer membrane
relatively slow and this is one of the reasons why Gram negative
bacteria have high antibiotic resistance
11/11/24 RSM & DGK 102
 Outer Membrane cont...
Note:
• Permeability of outer membrane varies
widely from one Gram negative species to
another
• Example; in Pseudomonas, the outer
membrane is 100 times less permeable than
the outer membrane of E. coli.
• Due to this, Pseudomonas are extremely
resistant to antibiotics
11/11/24 RSM & DGK 103
11/11/24 RSM & DGK 104
 Roles of outer membrane

• Has strong negative charge that helps evade phagocytosis and

action of complement
• Provides a barrier to antibiotics, lysozyme, detergents, heavy
metals, bile salts, digestive enzymes and certain dyes
• Permeability of porins allows passage of small mols.
• Specific channel proteins-allow specific substs like Vit B12, Fe,
nucleotides
• Provides attachment site for viruses and bacteriocins
• LPS component of outer membrane (O-polysaccharides) function
as antigen and useful for distinction of species
• A lipid part (lipid A) is toxic i.e. Endotoxins cause fever and shock

11/11/24 RSM & DGK 105

3. Lipopolysaccharides (LPS)
• Is composed of three (3) distinct units
1.A phospholipid called lipid A
2.A core polysaccharide of five sugars linked
through ketodeoxyoctulonic acid (KDO) to lipid A
3.An outer polysaccharide consisting of up to 25
repeating units of three five sugars
• Lipid A is responsible for toxic effects of LPS
• The outer polysaccharide is the important
surface antigen of bacterial cells called
“Somatic or O-antigen” that is used to identify
certain organism in clinical laboratory
11/11/24 RSM & DGK 106
 Endotoxin (LPS) structure: The antigen polysaccharide is
exposed to the exterior (up) of the cell where the lipid A faces
interiorly (down)
11/11/24 RSM & DGK 107
 LPS cont...
• Antigen specificity is conferred by the terminal
repeat units surrounding the cell by forming a layer
of hydrophilic polysaccharides
• Number of possible antigenic types in very great,
over 1000 are recognized in Salmonella alone!
• LPS is synthesized on cytoplasmic membrane &
transported to exterior position, and attached to the
outer membrane by hydrophobic bonds
• LPS is required for functioning of many membrane
proteins

11/11/24 RSM & DGK 108

 LPS cont...
• The polysaccharide core is similar in all
Gram negative species but each species
contains a unique repeat unit which is usually
linear trisaccharide or branched tetra or
pentasaccharides
• The negatively charged LPS molecules are
non covalently cross-bridged by divalent
cations, stabilizing the membrane and
provide barrier to hydrophobic molecules

11/11/24 RSM & DGK 109

Note:
• When the divalent cations are removed, e.g.
by chelating agents or when displaced by
polycationic antibiotics such as polymyxins
and aminoglycosides, the outer membrane is
rendered permeable to large hydrophobic
molecules
• LPS is extremely toxic to animals and is
referred to as the endotoxin of Gram
negative bacteria (firmly bound to cell surface
and released when the cells are lysed)
11/11/24 RSM & DGK 110
11/11/24 RSM & DGK 111
 LPS cont...
• LPS is responsible for features of diseases
such as fever and shock (especially
hypotension) caused by Gram negative
organisms
• LPS is called endotoxin because it is an
integral part of the cell wall, in contrast to
exotoxins which are freely released from the
bacteria.
• Pathologic effects of endotoxins are similar
irrespective of source of organism

11/11/24 RSM & DGK 112

 NOTE
• When growing bacteria, if the culture appears:-
– Mucoid = Smooth (lots of LPS or capsule)
– Dry = Rough (little LPS or capsule)
• Not all Gram negative bacteria have outer membrane LPS with
the discussed repeat units
• Outer membrane glycolipids of bacteria that colonize mucosal
surfaces such as Neisseria meningitidis, Neisseria gonorroeae,
Haemophilus influenzae, and Haemophilus ducrey have short
branched glycans whose structures resemble those of
glycosphingolipids of mammalian cell membrane, these glycans are
called Lipooligosaccharides (LOS)
– LOS show extensive antigenic and structural diversity even within
a single strain
– LOS is an important virulent factor
– Epitopes on LOS mimic host structures and may enable these
organisms to evade the immune response of the host
11/11/24 RSM & DGK 113
 Summary on LPS
• Functions
– Toxic; kills mice, pigs, humans
• G- septicemia; death due to LPS
– LPS is Pyrogenic; causes fever
• Example, DPT vaccination always causes fevers
– Adjuvant; stimulates immunity
• Heat Resistant;
• hard to remove
• Detection (all topical & IV products)
– Rabbits (measure fever)
– Horse shoe crab (Amoebocytes Lyse in presence of LPS)

11/11/24 RSM & DGK 114

 4. Periplasmic Space
• Is a space between inner and outer membrane (20-40% of cell volume)
• Contains the murein layer and a gel-like solutio of proteins
• The periplasmic proteins include:
– Binding proteins for specific substrates e.g. amino acids, sugars,
vitamins and ions
– Hydrolytic enzymes (e.g. alkaline phosphatase and 5’-nucleotidase)
that break down non transportable substrate into transportable ones
– Detoxifying enzymes, (e.g. β-lactamase and aminoglycoside-
phosphorylase) that inactivate β-lactam and aminoglycoside
antibiotics
– Highly branched polymers which play a role in osmoregulation

11/11/24 RSM & DGK 115

 Enzymes That Attack Cell
Walls
• The β1-4 linkage of the peptidoglycan backbone is
hydrolyzed by the enzyme, lysozyme, found in
animal secretions such as teas, saliva, nasal
secretions, and in egg white
• Gram positive bacteria treated with lysozymes in
low osmotic strength media lyse, but if osmotic
strength of the medium is raised to balance the
internal osmotic pressure of the cell, then, free
protoplasts are liberated

11/11/24 RSM & DGK 116

 • Outer membrane prevent access of lysozymes
unless disrupted by chelating agents (e.g. EDTA)
– In osmotically protected media, such cells
treated with EDTA and lysozymes form
spheroplasts that still possess remnants of the
complex Gram negative wall and the outer
membrane

11/11/24 RSM & DGK 117

 Enzymes That Attack Cell Walls
cont...
• Bacteria possess autolysins that attack
peptidoglycan
– E.g. glycosidases, amidases, and peptidases
– These autolytic enzymes are essential in cell
growth and division but most importantly in
dissolution of dead cells (Autolysis)
• Enzymes that degrade bacterial cell walls are
also found in protozoa and phagocytic cells of
higher animals

11/11/24 RSM & DGK 118

Protoplasts, Spheroplasts & L
forms

• Bacterial cell wall may be removed by;-

– Hydrolysis with lysozymes or
– By blocking peptidoglycan biosynthesis with
antibiotics such as Penicillins, cephalosporins etc.
• In osmotically protected media, such treatments liberate;-
– Protoplasts from Gram positive bacteria and
– Spheroplasts (Which retain outer membrane and
entrapped peptidoglycan) from Gram negative cells

11/11/24 RSM & DGK 119

 Protoplasts, Spheroplasts & L
forms
• If these cells are able to grow and divide, then, they
are L forms
• L forms are difficult to cultivate and usually require
a medium that is solidified with agar and with right
osmotic strength
• L forms are produced readily with penicillins than
with lysozymes
• Some L forms can revert to normal bacillary form
when inducing stimulus is removed ie are able to
resume normal cell wall synthesis

11/11/24 RSM & DGK 120

 Protoplasts, Spheroplasts & L forms
cont...
• Other L forms are stable and never revert ( factor determining
capacity to revert may be presence of residual peptidoglycan
which acts as a primer for its own biosynthesis)
• Some bacterial species produce L forms spontaneously
• Spontaneous or antibiotic induced formation of L forms in the
host (patient) may produce chronic infections
• The organism persists by becoming sequestered in protective
region of the body
• Because L forms infections are relatively resistant to antibiotics,
they present a special problem in chemotherapy
• Their reversion to bacillary form can produce relapses

11/11/24 RSM & DGK 121

 Specialized Structures Outside The Cell
Wall
Capsule, Glycocalyx & Slime layer
• The capsule is a gelatinous, condensed, well defined layer, closely
surrounding and covering the entire bacterium
• The capsule is composed of polysaccharide (except in Bacillus
anthracis whose capsule is composed of poly-D-glutamate)
• The polysaccharide polymer (or poly-D-glutamate) is synthesized
by bacteria when growing in their natural environment
• When the polymer forms a loose meshwork of fibrils extending
outward form the cell, it is called the Glycocalyx.
Note
• When masses of polymer are formed totally detached from cell but
in which cells may be entrapped; then the polymer is now called the
Slime Layer

11/11/24 RSM & DGK 122

 Capsule, Glycocalyx & Slime layer
cont...
• Sugar component of the polysaccharide forming
capsule, glycocalyx or slime layer vary from
one bacterial species to another and frequently
determine the Serologic type within species
• For example:
– There are 84 different serologic types of
Streptococcus pneumoneae which are distinguished
by the antigenic differences of the sugars in the
polysaccharide of the capsule

11/11/24 RSM & DGK 123

 Importance of Capsule
• Contributes to invasiveness of pathogic bacteria i.e.
determines virulence of many bacteria
– Encapsulated cells are protected from phagocytosis,
• The capsule limits ability of phagocytes to engulf
the bacteria
– Phagocytes engulf encapsulated cells only if the
encapsulated cell is coated with anticapsular antibody
– Variants of encapsulated bacteria that have lost ability
to produce a capsule are usually non pathogenic

11/11/24 RSM & DGK 124

Importance of Capsule cont...
• The capsule is important in specific
identification of an organism by using
antiserum against the capsular polysaccharide
– In presence of homologous antibody, the
capsule swells greatly
– The swelling phenomenon (quelling
reaction) is used to identify certain bacteria

11/11/24 RSM & DGK 125

 Importance of Capsule cont...

• Capsular polysaccharides are used as antigens

in certain vaccines because they are capable of
eliciting protective antibodies
– For example; the purified capsular
polysaccharides of 23 types of S. pneumoneae are
present in the current vaccine
• The capsule may play a role in adherence of
bacteria to human tissues which is an
important initial step in causing infection
11/11/24 RSM & DGK 126
 Importance of Glycocalyx
• Plays a role in adherence of bacteria to surfaces in their
environment, including cells of plant and animal hosts
– E.g. S. mutans adhere tightly to enamel by its glycocalyx
• Bacterial cell of the same or different species become
entrapped in the glycocalyx and form a layer known as
“plaque” on tooth surface. Acidic products excreted by those
bacteria cause dental carries
• Essential role of glycocalyx in this process and its formation
from sucrose explains the relationship between dental carries
and sucrose consumption by human population

11/11/24 RSM & DGK 127

 Flagella
Structure
• Bacterial flagella are long thread-like appendages composed
entirely of proteins
• Are organs of locomotion for bacteria possessing them
• Flagella are classified on the basis of their arrangement
a. Monotrichous
•single polar (at one end) flagellum
b. Lophotrichous
•Multiple polar flagella
c. Amphitrichous
•both ends
d. Peritrichous
•Flagella distributed all over the entire bacterial cell

11/11/24 RSM & DGK 128

 Flagella Arrangement

11/11/24 RSM & DGK 129

 Flagella cont...

• Bacterial flagella are made up of protein subunits called

flagellin which aggregate to form a helical structure
• If flagella are removed by mechanical agitation of a
suspension of bacteria, new flagella are rapidly formed
and motility is restored within 3-6 minutes
• Flagellins of different bacterial species differ from one
another in primary structure
• Flagellins are highly antigenic (H antigens) and some
of the immune response are directed against these
proteins

11/11/24 RSM & DGK 130

 Flagella cont...
• The flagellum is attached to bacterial cell body
by a complex structure consisting of a hook
and a basal body
• The hook is short and curved that acts like a
universal joint between the motor in the basal
structure and the flagellum (see next slide)
• The basal body has a set of rings, one pair in
Gram positive bacteria and two pairs in Gram
negative bacteria

11/11/24 RSM & DGK 131

11/11/24 RSM & DGK 132
 Flagella Structure

11/11/24 RSM & DGK 133

 Flagella cont...
Motility
• Bacterial flagella are semirigid helical rotors to which the cell
imparts a spinning movement
• Rotation is driven by flow of protons into the cell down the
gradient produced by primary proton pump shown in previous
slide
• In absence of metabolic energy source rotation is driven by
proton motive force generated by ionophores
– Bacteria living in alkaline environments (alkalophiles)
use energy from sodium ion gradient (not proton gradient)
• All components of flagellar rotor are located in cell envelope

11/11/24 RSM & DGK 134

11/11/24 RSM & DGK 135
 Mono- or Lophotrichorus

11/11/24 RSM & DGK 136

 Pili (Fimbriae)
• Are rigid surface appendages found in many Gram negative
bacteria
• Are shorter and finer than flagella
• Are composed of structural protein subunits called pilins
• Some pili contain single type of pilin, other more than one
• Minor proteins located tips of pili are responsible for
attachment properties
• There are two classes of pili:-
– Ordinary pili
• For adherence of symbiotic bacteria to host cells
– Sex pili
• For attachment of donor and recipient cells in bacterial
conjugation

11/11/24 RSM & DGK 137

 Pili

11/11/24 RSM & DGK 138

 Pili (Fimbriae) cont...
• Ordinary pili contribute to virulence of some
pathogenic bacteria (facilitate adhesion to
host)
• In enteropathogenic E. coli strains, both
enterotoxins and pili are genetically
determined by transmissible plasmids
• In Streptococci, fimbriae are the site of major
surface antigen, the M protein
• Lipoteichoic acid associated with fimbriae is
responsible for adherence of group A
streptococci to host epithelia cells
11/11/24 RSM & DGK 139
 The Cell Envelope

• Includes layers surrounding prokaryotic

cell
• Structure and organization of cell
envelope differ in Gram positive and
Gram negative bacteria

11/11/24 RSM & DGK 140

 Gram Positive Cell Envelope
• Is relatively simple, consisting of 2 to 3 layers
• The cytoplasmic membrane
• The peptidoglycan layer (thick) and
• Capsule (in some bacteria)

Gram Negative Cell Envelope

• Is highly complex, multilayered structure
• Includes;
• Cytoplasmic membrane which is also called the inner membrane in
Gram negative bacteria
• Single planar sheet of peptidoglycan (thin), to which a complex layer
(outer membrane) is anchored
• Capsule (outer most layer)
– The space between the inner and outer membrane is called the
Periplasmic space

11/11/24 RSM & DGK 141

Structural Comparison of Gram
+ve & Gram –ve cell Envelopes

11/11/24 RSM & DGK 142

 Endospores
• Members of several bacterial genera are capable of
forming endospores
• Two most common are Gram positive rods
– The obligately aerobic genus Bacillus and
– The obligately anaerobic genus Clostridium
• Other bacteria that form endospores are the Gram positive
coccus, Sporosarcina and Coxiella burnetii, the agent of
Q fever
• Organisms forming spores undergo a cycle of
differentiation in response to environment conditions
• When nutrients are depleted, each cell forms a single
internal spore that is liberated when the mother cell
undergoes autolysis

11/11/24 RSM & DGK 143

Endospores cont...
• The spore is a resting cell, highly resistant to desiccation,
heat, and chemical agents
• When returned to favorable nutritional conditions and
activated, the spores germinate to produce a single
vegetative cell
• Position of endospore differs among bacterial species and
is useful in identification.
• Main types within the cell are:
– Terminal (e.g. Clostridium tetani),
– Subterminal (e.g. Bacillus subtilis ) and
– Centrally placed endospores (e.g. Bacillus cereus )
11/11/24 RSM & DGK 144
 • Terminal endospores are seen at the poles of cells,
central are more or less in the middle and
Subterminal endospores are between these two
extremes
• Lateral endospores are seen occasionally.
• Sometimes the endospore can be so large the cell
can be distended around the endospore,
• this is typical of Clostridium tetani.

1, 4 –Central,
2,3,5 –Terminal,
6 –Lateral endospore

11/11/24 RSM & DGK 145

 Sporulation or
Sporogenesis
• Sporulation is the process by which spores are formed
• Sporulation begins when nutritional conditions become
unfavorable
– Depletion of nitrogen or carbon source (or both) is
the most significant factor
• Sporulation occurs massively in cultures that have
terminated exponential growth as a result of depletion
• Sporulation involves production of many new structures,
enzymes, and metabolites along with disappearance of
many vegetative cell components

11/11/24 RSM & DGK 146

 • The sequence of events in sporulation is highly
complex:
– Differentiation of a vegetative cell of B
subtilis into an endospore takes about 7
hours under lab conditions
• Different morphologic & chemical events
occur at sequential stages of the process
• Seven stages have been identified
• During the process some bacteria release
peptide antibiotics which may play a role in
regulating sporogenesis
11/11/24 RSM & DGK 147
• Morphologically,
– Sporulation begins with formation of an axial filament
• The process continues with an infolding of the membrane to
produce a double membrane structure whose facing surfaces
correspond to the cell wall synthesizing surface of the cell
envelope
• The growing points move progressively toward the pole of the
cell to engulf the developing spore
• The two spore membrane now engage in active synthesis of
special layers that will form the cell envelope:
– The spore wall and the cortex, lying between the facing
membranes and
– The coat and exosporium, lying outside the facing
membranes
11/11/24 RSM & DGK 148
11/11/24 RSM & DGK 149
 Stages of Sporulation
11/11/24 RSM & DGK 150
 Properties of Endospores
The core
• Is the spore protoplast containing a complete
nucleus (chromosome), all of components of
protein-synthesizing apparatus, and an energy
generating system based on glycolysis
• Energy for germination is stored as 3-
phosphoglycerate rather than as ATP
• Heat resistance of spore is due (in part) to their
dehydrated state and presence of large amount (5-
15% of spore dry weight) of calcium dipicolinate
which is formed from an intermediate of lysine
biosynthetic pathway (discussed later)
11/11/24 RSM & DGK 151
Spore Wall
• The innermost layer surrounding the inner spore membrane is
called the spore wall
• It contains normal peptidoglycan and become the cell wall of
the germinating vegetative cell
Cortex
• Is the thickest layer of the spore envelope
• Contains unusually type of peptidoglycan, with many fewer
cross-links than are found in cell wall peptidoglycan
• Cortex peptidoglycan is extremely sensitive to lysozyme
– Autolysis of the cortex plays a role in spore germination
Coat
• Composed of keratin-like protein containing many
intermolecular disulfide bonds
• Impermeable offer relative resistance to antibacterials
11/11/24 RSM & DGK 152
 Exosporium
• Is a lipoprotein membrane containing some
carbohydrate
Note:
• Alkylating agents such as ethylene oxide, are effective
against endospores
Germination
• The germination process occurs in three stages:-
» Activation
» Initiation and
» Outgrowth

11/11/24 RSM & DGK 153

 Activation
• Most endospores can not germinate immediately after
being formed
• The period during which spores are in resting condition
is called spore dormancy
• Spores can germinate after they have rested for several
days or are first activated;
» In nutritionally rich medium or
» By agent that damages the spore coat
• Agents that overcome spore dormancy include:-
» Heat,
» Abrasion
» Acidity and
» Compounds containing free sulfhydryl groups

11/11/24 RSM & DGK 154

 Initiation
• Once activated, spore initiates germination if
environmental conditions are favorable
– Different species have evolved receptors that recognize
different effectors as signaling a rich medium
• Initiation may be triggered by L-alanine in one species
and by adenosine in another species
– Binding of effector activates an autolysin that
rapidly degrades the cortex peptidoglycan
– Water is then taken up, calcium dipicolinate is
released and variety of spore constituents are
degraded by hydrolytic enzymes

11/11/24 RSM & DGK 155

Outgrowth
• Degradation of the cortex and outer layers
results in emergence of a new vegetative cell
consisting of the spore protoplast with its
surrounding wall
• A period of active biosynthesis follows;
– This period terminates in cell division
– Is called outgrowth
– Outgrowth requires a supply of all nutrients
essential for cell growth

11/11/24 RSM & DGK 156

 Pharmaceutical Importance of bacterial
spores
• Are structures resistant to:-
– Heat, irradiation, cold, chemicals
– Boiling >1 hr, still viable
– Survive in undercooked food, hence produce toxins
and diseases
• Bacillus stearothermophilus -spores
– Used for quality control of heat sterilization equipment
• Bacillus anthracis - spores
– Used in biological warfare

11/11/24 RSM & DGK 157

 Prokaryotic vs. Eukaryotic

11/11/24 RSM & DGK 158

11/11/24 RSM & DGK 159
 Study questions

Between Gram negative and Gram positive

bacteria,
– Which group has a thicker cell wall? Why?(-ve)
– Which group has endotoxins (LPS)?(-ve)
– Which group has teichoic acid?(+ve)
– Which are more sensitive to Penicillin? Why?(-ve)
– Alcohol/Acetone affects which group more? Why
– With the aid of diagram describe types of bacterial
flagela and fimbriae.

11/11/24 RSM & DGK 160