B 484-Microbiology

Lecture 2: Bacteria Structure and Function

Dr Liteboho D. Maduna

Department of Biology

Copyright © McGraw-Hill Education. Permission required for reproduction or display. 1

Bacterial Shapes and Arrangements

• Most bacteria function as independent single-celled, unicellular organisms.

– Some act as a group in colonies or biofilms.
– Some communicate through nanowires.
• Bacteria have an average size of 1 μm (micron).
– Cocci: circumference of 1 μm
– Rods: length of 2 μm and a width of 1 μm
• Pleomorphism: variation in the size and shape of cells of a single species
due to nutritional and genetic differences
Bacterial Shapes and Arrangements

1. Arrangement of Cocci (s. coccus): spheres

• Single
• Diplococci: pairs
• Ex. Neisseria gonorrhoeae causes gonorrhoea (sexually transmitted
infection)
• Tetrads: groups of four Ex. Micrococcus luteus
• Staphylococci or micrococci: irregular clusters
• Ex. Staphylococcus aureus causes skin infections e.g Boils
• Streptococci: chains
• Ex. Streptococcus pyogenes causes tonsilitis (strep throat)
Bacterial Shapes and Arrangements

2. Arrangement of bacilli: rods

– coccobacilli – very short rods
– Ex. Haemophilus influenza – causes otitis media,
respiratory infections, and meningitis in children)
– Diplobacilli: pair of cells with ends attached
– Streptobacilli: chain of several cells
– Palisades: cells of a chain remain partially attached by a
small hinge region at the ends
– Ex. Vibrio cholerae
• Spirilla: occasionally found in short chains
• Spirochetes: rarely remain attached after cell division
Bacterial Shapes and Arrangements

3. Arrangement of spirochaete
– Spiral-shaped, occasionally found in short chains
– Moves by spinning around its long axis
– Example: Treponema pallidum – causative agent of
syphilis
4. Arrangement of Vibrios
• Curved or comma-shaped bacilli
• May occur singly or in pairs (gull-wing morphology
like Campylobacter jejuni)
• Campylobacter jejuni which causes diarrhea brought
about by poultry (chicken)
Cell Wall Deficient Bacteria

▪ Pleomorphic: exists in a variety of

shapes
▪ Mycoplasma genitalium cause sexually
transmitted infections
▪ Mycoplasma pneumoniae (cause of
atypical pneumonia), and Ureaplasma
urealyticum (cause of UTI)
Bacterial Cell Organization Common Features

▪ The general cellular organization of a bacterial cell can be represented with

this flowchart

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Structures common to all bacterial cells
• All bacterial cells possess:
– Cell membrane
– Cytoplasm
– Ribosomes
– Cytoskeleton
– One (or a few) chromosome(s)
• Most bacterial cells possess:
– Cell wall
– A surface coating called a glycocalyx
The Structure of the Bacterial Cell (cont’d)

▪ Some, but not all bacterial cells possess:

• Flagella, pili, and fimbriae
• An outer membrane
• Plasmids
• Inclusions
• Endospores
• Intracellular membranes
 The Cell Envelope

▪ Cell wall is strong, rigid structure that prevents cell lysis

▪ Lies outside the cytoplasm
▪ Composed of two or three basic layers:
• Cell wall; architecture distinguishes two main types of bacteria
• Gram-positive
• Gram-negative
• Plasma membrane
• Outer membrane (in some bacteria)
The Cell Wall
Functions
1. It helps determine the shape of the bacterium.
• Almost all bacteria have one
2. It provides strong structural support to keep the bacterium from bursting
or collapsing under osmotic pressure.
• Certain drugs target the cell wall, disrupting its integrity and causing cell lysis
(disintegration or rupture) of the cell.
3. Helps protect from toxic materials
4. May contribute to pathogenicity
Structure of the Cell Wall
▪ The bacterial cell wall owes its strength
to a layer composed of peptidoglycan
(murein)
▪ Peptidoglycan is a complex of
alternating amino-sugar and peptide
structure important for cell wall stability
and shape.
– two alternating sugars
• N- acetylmuramic acid (NAM)
• N-acetylglucosamine (NAG)
– alternating D- and L- amino acids
 The peptidoglycan layer

▪ Backbone: alternating N- acetylglucosamine (NAG) and N-acetylmuramic acid

(NAM) connected by β1 to 4 linkages (Glycan Chain)
▪ Tetrapeptide side chains attached to NAM
▪ Identical Peptide Cross bridges connecting the tetrapeptide chains.
 Gram-Positive Cell Walls

▪ Composed primarily of thick, homogenous sheet of

peptidoglycan
• 20 – 80 nm in thickness
▪ May also contain teichoic acids (negatively
charged)
• may account for up to 50% of the dry weight of the wall
and 10% of the dry weight of the total cell.
• help maintain cell envelope and enlargement
• protect from environmental substances
• may bind to host cells
▪ Some Gram-positive bacteria have layer of proteins
on surface of peptidoglycan
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The Gram-Positive Cell Wall

▪ Gram-positive cell wall has thick peptidoglycan layer

Gram-positive cell walls

▪ Teichoic acid is a polymer of ribitol or glycerol and phosphate

embedded in the peptidoglycan sheath.
▪ Functions in cell wall maintenance and enlargement during cell
division. They also move cations into and out of the cell and
stimulate a specific immune response
Gram-positive cell envelope

▪ The teichoic acids are covalently connected to either the peptidoglycan

itself or to plasma membrane (called lipoteichoic acid).
Gram-Negative Cell Walls

▪ More complex than Gram-positive walls

▪ Consist of a thin layer of peptidoglycan
surrounded by an outer membrane
• Peptidoglycan is ~5-10% of cell wall weight
▪ Outer membrane composed of
• Lipids,
• Lipoproteins, and
• Lipopolysaccharide (LPS)
▪ No teichoic acids

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Gram-negative cell walls

▪ The outer membrane is bilayered in structure.

▪ Consists of lipopolysaccharides (LPS) and lipoproteins.
▪ The lipopolysaccharide (also known as endotoxin)
Lipopolysaccharide (LPS)

▪ The LPS, also known as the endotoxin because

they are extremely toxic to animals
▪ Composed of lipid molecules bound to
polysaccharides
▪ LPS consist of three parts:
1. lipid A,
2. the core polysaccharide,
3. the O sidechain.
▪ The lipid A component is embedded in the outer
leaflet of the membrane anchoring the LPS.
 LPS Structure

▪ (1) Lipid A is the core structure of LPS. Consists of phosphorylated glucosamine disaccharide
units to which are attached a number of long-chain fatty acids.
– The structure and chemical compositions of lipid A are similar in nearly all Gram-negative
bacteria.
– So the pathophysiologic effects of LPS (endotoxin) are similar regardless of their bacterial
origin.
 LPS Structure

▪ (2) The polysaccharide core: similar in all gram-negative species that have
LPS.
– Includes two characteristic sugars, ketodeoxyoctanoic acid (KDO) and a heptose.
▪ (3) O side chain: each species, however, contains a unique repeat unit called
the O antigen.
– The repeat units are usually linear trisaccharides or branched
tetra/pentasaccharides.
– The fine chemical structure of the O antigen results in a large number of
antigenic variants useful in bacterial typing (e.g., detailed differentiation of
salmonella type)
Lipoprotein

▪ Molecules of an unusual lipoprotein cross-link the outer membrane and

peptidoglycan layers.
▪ The lipoprotein contains 57 amino acids, representing repeats of a 15-
amino-acid sequence; it is peptide-linked to diaminopimelic acid
residues of the peptidoglycan tetrapeptide side chains.
▪ The lipid component, consisting of a diglyceride thioether linked to a
terminal cysteine, is noncovalently inserted in the phospholipid bilayer.
▪ Its function is to stabilize the outer membrane and anchor it to the
peptidoglycan layer.
Structures associated with Gram-positive and Gram-negative cell walls.
Comparison of Gram-Positive and Gram-Negative Cell Walls
Gram Stain Reaction

▪ This is the most extensively used stain to differentiate between different

types of bacteria
▪ Most bacteria are classified as Gram-positive or Gram-negative
according to their response to the Gram-staining procedure.
▪ Gram staining procedure was developed by Hans Christian Gram in
1884.
▪ Gram-positive bacteria stained purple, retain a complex of crystal
violet (a purple dye) and iodine after a brief wash with alcohol or acetone.
▪ Gram-negative bacteria colored pink or red do not retain the dye–
iodine complex and become translucent, but they can then be
counterstained with safranin (a red dye).
 Gram Stain Procedure

1. Crystal violet
▪ First, crystal violet is added to the cells in a
smear. It stains them all the same purple
color.
2. Gram’s iodine
▪ Then, the mordant, Gram’s iodine, is
added. This is a stabilizer that causes the
dye to form large complexes in the
peptidoglycan meshwork of the cell wall.
▪ The thicker gram-positive cell walls are able
to more firmly trap the large complexes
than those of the gram-negative cells.
 Gram Stain Procedure

3. Alcohol
▪ Application of alcohol dissolves lipids in the outer membrane and
removes the dye from the peptidoglycan layer—only in the gram-
negative cells.
4. Safranin (red dye)
▪ Because gram-negative bacteria are colorless after decolorization, their
presence is demonstrated by applying the counterstain safranin in the
final step
The steps in a Gram stain
 Gram-Positives Gram Stain Gram-negatives

Gram-positive cocci in clusters (Staphylococci) Gram-negative rods

Gram-positive rods

Gram-positive Cocci in
chains (Streptococci) Gram-negative cocci
Mechanism of Gram Stain Reaction

1. Gram stain reaction due to nature of cell wall

2. Crystal violet stains inside of cell, not cell wall
3. Gram-positive cell wall prevents crystal violet–iodine complex from
being washed out
▪ Constriction prevents loss of crystal violet during decolorization step
▪ Shrinkage of the pores of peptidoglycan layer of Gram-positive cells
3. Thinner peptidoglycan layer and larger pores of Gram-negative bacteria
does not prevent loss of crystal violet

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Mycoplasmas and Other Cell-Wall- Deficient Bacteria
▪ Mycoplasma bacteria have no cell wall, which contributes to varied shapes (pleomorphic)
▪ Mycoplasma cell membrane is stabilized by sterols and is resistant to lysis
• Naturally lack a cell wall
• Very small bacteria (0.1 to 0.5 µm) !!!
• Range in shape from filamentous to coccus !!!
• Live in isotonic environments !!!
• Cannot be grown on artificial media
• Found in many habitats
▪ Important medical species:
• Mycoplasma genitalium (causes sexually transmitted infections)
• Mycoplasma pneumonia
Scanning electron micrograph of Mycoplasma genitalium
Differences in Cell Envelope Structure

• Outer membrane of gram-negative bacteria contributes an extra barrier:

– Impervious to certain antimicrobial chemicals e.g antibiotics
– More difficult to kill or inhibit than gram-positive bacteria.
• Alcohol-based compounds dissolve lipids in the outer membrane:
– Alcohol swabs used to cleanse the skin before certain medical procedures
• Treatment of infections caused by gram-negative bacteria requires drugs
that can cross the outer membrane.
 Cytoplasmic Membrane Structure
• Absolute requirement for all living organisms
• A lipid bilayer with proteins embedded.
• Regulates transport of nutrients and wastes
• Selectively permeable: special carrier mechanisms for passage of most molecules
• Serves as a site for:
– Energy reactions
– Nutrient processing
– Synthesis
• Interacts with external environment
• receptors for detection of and response to chemicals in surroundings
• transport systems
• metabolic processes
The Cytoplasmic Membrane

▪ Cytoplasmic membrane defines boundary of cell

• Phospholipid bilayer embedded with proteins
• Hydrophobic tails face in; hydrophilic tails face out
• Serves as semipermeable membrane
• Proteins serve numerous functions
• Selective gates
• Sensors of
environmental
conditions
• Fluid mosaic model:
proteins drift about in
lipid bilayer
The Cytoplasmic Membrane

▪ Cytoplasmic membrane defines boundary of cell (continued…)

• Bacteria and Archaea have same general structure of cytoplasmic
membranes
• Distinctly different lipid compositions
• Lipid tails of Archaea are not fatty acids and are connected
differently to glycerol
Permeability of Lipid Bilayer
▪ Cytoplasmic membrane is selectively permeable
• O2, CO2, N2, small hydrophobic molecules, and water pass freely
• Some cells facilitate water passage with aquaporins
• Other molecules must be moved across membrane via transport
systems
Permeability of Lipid Bilayer

▪ Simple Diffusion
• Movement from high to
low concentration
• Speed depends on concentration
▪ Osmosis
• Diffusion of water across selectively
permeable membrane due to
unequal solute concentrations
• Three terms:
– Hypertonic
– Isotonic
– Hypotonic
 Cytoplasmic Membrane and Energy Transformation

▪ Electron Transport Chain embedded in membrane

• Critical role in converting energy into ATP
• Eukaryotes use membrane-bound organelles
• Use energy from electrons to move protons out of cell
• Creates electrochemical gradient across membrane
• Energy called proton motive force
• Harvested to drive
cellular processes
including ATP
synthesis and
some forms of
transport, motility
Transport of Small Molecules Across Cytoplasmic Membrane

▪ Most molecules must pass through proteins functioning as selective gates

• Termed transport systems
• Proteins may be called permeases, carriers
• Membrane-spanning
• Highly specific: carriers transport certain molecule type
Transport of Small Molecules Across Cytoplasmic Membrane

▪ Facilitated diffusion is a form of passive transport

• Movement down gradient; no energy required
• Not typically useful in low-nutrient environments
▪ Active transport requires energy
• Movement against gradient
• Two main mechanisms
• Use proton motive force
• Use ATP (ABC transporter)
▪ Group Translocation
• Chemically alter compound
• Phosphorylation common
– Glucose, for example
Transport of Small Molecules Across Cytoplasmic Membrane

▪ Types of transport systems

Transport of Small Molecules Across Cytoplasmic Membrane

▪ Protein secretion: active movement out of cell

• Examples: extracellular enzymes, external structures
• Proteins tagged for secretion via signal sequence of amino acids
Cytoplasm

▪ Contents of the Cell Cytoplasm

• Gelatinous solution
• Site for many biochemical and synthetic activities
• 70%-80% water
• 20% - 30% nutrients, proteins, and genetic material (i.e., chromatin, ribosomes,
macromolecular storage granules, and actin strands)
The Nucleoid

▪ Not membrane bound

▪ Location of chromosome and associated
proteins
• Circular or linear double strands of DNA
▪ Supercoiling and nucleoid proteins
(different from histones) aid in folding
Plasmids
▪ Extrachromosomal pieces of DNA
• found in bacteria, archaea, some fungi
• usually small, closed circular DNA
molecules
▪ Exist and replicate independently of
chromosome
▪ Often confer protective traits such as
drug resistance or the production of
toxins and enzymes
 Ribosomes
▪ Site of protein synthesis
▪ Composed of rRNA and proteins
▪ Consist of a large and small subunit
• Small subunit: 30S
• Large subunit: 50S
• Large and small subunits together: 70S
• Archaeal ribosome: 70S
• Eukaryotic ribosome: 80S
Components Outside of the Cell Wall
▪ Outermost layer in the cell envelope
▪ Glycocalyx
1. capsules and slime layers
2. S layers
Functions
▪ Formed by many pathogenic bacteria- protect the bacteria against
phagocytes
▪ Aid in attachment to solid surfaces
• e.g., biofilms in plants and animals

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 S Layer and Glycocalyx

• S layer:
– Single layers of thousands of copies of a single protein linked together like chain
mail.
– Only produced when bacteria are in a hostile environment
• Glycocalyx:
– Coating of repeating polysaccharide or glycoprotein subunits
– Slime layer: loose, protects against loss of water and nutrients
– Capsule: more tightly bound, denser, produce a mucoid characters to colonies
on agar
Capsules

▪ Usually composed of polysaccharides

▪ Well organized and not easily removed from cell
▪ Visible in light microscope
▪ Protective advantages
• resistant to phagocytosis
• protect from desiccation
• exclude viruses and detergents

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Capsule

▪ Contributes to the invasiveness of pathogenic

bacteria encapsulated cells are protected from the
host cells phagocytosis.
▪ E.g When S. pneumoniae lacks a capsule, it is
destroyed easily and does not cause disease,
whereas the capsulated variant quickly kills mice.
▪ Capsules contain water and can protect against
desiccation. They exclude viruses and most
hydrophobic toxic materials such as detergents.
▪ Encapsulated bacteria are very pathogenic!!
Specialized Functions of the Glycocalyx

• Capsules:
– Formed by many pathogenic bacteria
– Have greater pathogenicity
– Protect against phagocytosis
• Biofilms:
– Plaque on teeth protects bacteria from becoming dislodged.
– Responsible for persistent colonization of catheters, pacemakers, and other
implanted medical devices.
 Appendages for Attachment and Mating
• Fimbria/fimbriae:
– Small, bristle-like fibers sprouting off the surface of many
bacterial cells (up to 1,000/cell)
– Allow tight adhesion between fimbriae and epithelial cells,
allowing bacteria to colonize and infect host tissues.
• Sex Pilus/pili:
– longer, thicker, and less numerous (1-10/cell)
– Used in conjugation between bacterial cells.
– genes for formation found on plasmids
Bacterial Flagella

▪ Threadlike, locomotor appendages extending outward from plasma

membrane and cell wall
▪ Thin, rigid protein structures that cannot be observed with bright-field
microscope unless specially stained
▪ Functions
• motility and swarming behavior
• attachment to surfaces
• may be virulence factors
▪ Pattern of flagellation varies

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Flagella

▪ Three parts
• Filament
• Hook
• Basal body
Patterns of Flagella Distribution

▪ Polar arrangement – flagellum attached at

one or both ends of cell
a) Monotrichous – single flagellum (polar)
b) Lophotrichous – small cluster of flagella
emerging at one or both ends (polar)
c) Amphitrichous – single flagellum, small
bunches or tufts of flagella emerging from
two sites
▪ Peritrichous arrangement – spread over
entire surface of cell
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 Lophotrichous

Monotrichous
Petrichous Amphitrichous
Chemotaxis

▪ Bacteria sense chemicals and move accordingly

• Nutrients may attract, toxins may repel
▪ Movement is series of runs and tumbles
▪ Other responses observed
• Aerotaxis
• Magnetotaxis
• Thermotaxis
• Phototaxis
The Bacterial Endospore
▪ Endospores (spores) are small, metabolically dormant cells
with a thick, multi-layered coat. Various locations within the
cell
▪ Formed extracellularly by members of the genera Bacillus and
Clostridium
▪ Highly resistant to adverse environmental conditions.
• heat
• radiation
• chemicals
• desiccation
▪ Spores are formed in response to limitations of nutrients by a
process of sporulation.
Sporulation

▪ Sporulation triggered by carbon, nitrogen

limitation
• Starvation conditions begin 8-hour process
• Endospore layers prevent damage
• Exclude molecules (e.g., lysozyme)
• Cortex maintains core in dehydrated state, protects
from heat
• Core has small proteins that bind and protect DNA
• Calcium dipicolinate seems to play important
protective role
▪ Germination triggered by heat, chemical
exposure
 The Medical Significance of Bacterial Endospores

▪ Spores also have an important role in the epidemiology of certain human

diseases
▪ Bacillus anthracis: agent of anthrax
▪ Clostridium tetani: cause of tetanus
▪ Clostridium perfringens: cause of gas gangrene
▪ Clostridium botulinum: cause of botulism
▪ Clostridium difficile: “C. diff,” a serious gastrointestinal disease
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