B.E Pruitt & Jane J.

Stein

Chapter 02
The Prokaryote: Bacteria

PowerPoint® Lecture Slide Presentation

REFERENCES and FIGURES

Tortora GJ, Funke BR, Case CL, 2007, Microbiology

an Introduction, 9th edition, Benjamin Cummings, San
Francisco, CA 94111, USA

Madigan MT, Martinko JM, 2006, Brock Biology of

Microorganisms, 11th edition, Pearson Education Inc.,
USA

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 Prokaryotic Cells

• Comparing Prokaryotic and Eukaryotic Cells

• Prokaryote comes from the Greek words for
prenucleus.
• Eukaryote comes from the Greek words for
true nucleus.

Comparison between prokaryotes and eukaryotes

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 Size, shape and arrangements

• Average size: 0.2-2.0 µm diameter, 2-8 µm in length

• Basic shapes:

Size, shape and arrangements

• Unusual shapes
• Star-shaped (Stella)
• Square (Halophilic archaea, Haloarcula)
• Most bacteria are monomorphic
• A few are pleomorphic (Rhizobium, Corynebacterium)

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 Structure of bacterial cell

Glycocalyx
• Composed of polysaccharide, polypeptides, or both
• A capsule is neatly organized
• A slime layer is unorganized & loose
• Capsules prevent phagocytosis (B. anthracis)
• EPS allows cell to attach and colonize (Klebsiella);
biofilm formation
• Barrier for biocides, disinfectants, AB
• Source of nutrition (S. mutans)
• Protects against dehydration
• Inhibit the movement of nutrients

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 Flagella
• Semirigid, helical structure
• 3 basic parts
• Gram (-) 2 pair of rings
• Gram (+) inner pair
• Motility
• Taxis (attractant, repellent)
• Chemotaxis
(O2, ribose, galactose)
• Phototaxis
• Receptor in various location
• Flagella proteins are H
antigens

Flagella arrangement

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 Axial filaments

• Endoflagella
• In spirochetes
(Treponema pallidum)
• Anchored at one end of a
cell
• Rotation of the filaments
cause cell to move

Axial filaments

Endoflagella: bundle of fibrils that arise

at the ends of the cell beneath an outer
sheath and spiral around the cell

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 Fimbriae and Pili

• Fimbriae allow
attachment (biofilm)
• Few or several
hundred per cell
• N. gonorrhoeae
• Pili are used to
transfer DNA from
one cell to another
• One or two per cell
• Conjugation pili
• Mediated by plasmids

Cell wall
• Complex, semirigid structure
• Prevents osmotic lysis (water)
• Maintain shape of the cell and serves as a point
anchorage for flagella
• Contributes in some species to cause disease
• Made of peptidoglycan (in bacteria); site action of AB
• Compared to eukaryote’s cell wall, differ in
chemically, are simpler and structure and less rigid

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 Peptidoglycan (murein)
• Polymer of disaccharide
N-acetylglucosamine (NAG) & N-acetylmuramic acid
(NAM); murus means wall
• Linked in row of 10 to 65 sugars to form “backbone”
• Linked by polypeptides
• tetrapeptides side chain (4 amino acids) and peptides
cross-bridge

Cell wall

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 Gram (+) and (-) cell wall

Gram-positive cell wall

Gram-negative cell wall

Gram-positive cell wall

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 Gram-positive cell wall

• Forming thick and rigid structure

• Teichoic acids: negative charge, regulate the
movement of cations into and out of the cell
• Assume in cell growth, preventing excessive wall
breakdown and possible cell lysis
• Antigenic specificity, identification of bacteria
• Resistance to physical disruption

• In acid-fast cells, contains mycolic acid (60%);

thin layer peptidoglycan; held together by
polysaccharide
• Acid-fast cells: Mycobacterium and Nocardia

Gram-negative cell wall

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 Gram-negative cell wall

• Periplasm: contains hydrolytic enzymes, binding

proteins, chemoreceptors
• Outer membranes: strong negative charge, helps
evading phagocytosis and action of complement
• Protection to ABs, digestive enzymes (lysozyms,
detergents, heavy metals, bile salts, and certain dyes)
• Porins (protein): form channels (hydrophilic low-MW)
through membrane, e.g. nucleotides, disaccharides,
peptides, amino acids, vitamin B12 and iron
• O polysaccharides: antigens, determine the species
• Lipid A: endotoxins
• Penicillin vs gram negative: outer membranes, barrier

Characteristics of Gram (+) and (-)

CHARACTERISTIC GRAM POSITIVE GRAM NEGATIVE
Gram Reaction Retain crystal violet dye and Can be decolorized to accept
stain dark violet or purple counterstain (safranin) and
stain pink
Peptidoglycan Layer Thick (multilayered) Thin (single-layered)
Teichoic Acid Present in many Absent
Periplasmic Space Absent Present
Outer Membrane Absent Present
Lipopolysaccharide (LPS) Content Virtually none High
Lipid and Lipoprotein Content Low (acid-fast bacteria have High (due to presence of outer
lipids linked to peptidoglycan) membrane)
Flagellar Structure 2 rings in basal body 4 rings in basal body
Toxins Produced Primarily exotoxins Primarily endotoxins
Resistance to Physical Disruption High Low
Cell Wall Disruption by Lysozyme High Low (requires pretreatment to
destabilize outer membrane)
Susceptibility to Penicillin And Sulfonamide High Low
Susceptibility to Streptomycin, Low High
Chloramphenicol and Tetracycline
Inhibition of Basic Dyes High Low
Susceptibility to Anionic Detergents High Low
Resistance to Sodium Azide High Low
Resistance to Drying High Low

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 Atypical cell walls

• Genus Mycoplasmas
• Lack or very little material of cell walls
• Smallest bacteria that can grow and reproduce
outside living host cells
• Pass through 0,2 µm membrane filters
• Sterols in plasma membrane: help protect from lysis
• Starting materials requiring special attention
• Cell cultures for the production of veterinary
vaccines
• Cell substrates for the production of vaccines for
human use

Plasma membrane

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 Plasma membrane (inner membrane)
• Consist primarily phospholipids bilayer and protein
• Peripheral proteins
• catalyze chemical reaction and mediator of changes
in membrane shape during movement
• Integral proteins (transmembrane proteins): pore
• Selective permeability
• Simple sugars, O2, CO2, non polar substances
• Ions penetrates the membrane slowly
• Break down of nutrients and the production of energy
(chromatophores or thylakoids)
• Mesosomes ??? artifacts, not true cell structures
• Medically targeted: alcohols, QA, polymyxins

Plasma membrane

Rhodospirillum rubrum, a purple (nonsulfur) bacterium

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 Movement across membranes
• Passive process: movement of substance from ↑ [ ]
→ ↓ [ ], move with the gradient [ ], without ATP
• Simple diffusion: Movement of a solute from ↑ [ ]
→ ↓ [ ], equilibrium, small molecules (O2, CO2)
• Facilitative diffusion: Solute combines with a
transporter protein (permease) in the membrane
• Osmosis: movement of water across a selectively
permeable membrane from ↑ [ ] → ↓ [ ], isotonic
hypotonic, hypertonic solution
• Most bacteria live in hypotonic solution and
swelling contained by the cell wall
• Most bacteria produce enzymes that can break down
large molecules: extracellular enzymes, released by
bacteria into the surrounding medium

Movement across membranes

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 Movement across membranes

Movement across membranes

• Active process: movement of substance from ↓ [ ]

→ ↑ [ ], move against the gradient [ ], with ATP
• Active transport of substances requires a
transporter protein and ATP.
• Group translocation of substances requires a
transporter protein and phosphoenolpyruvic acid
(PEP)
• Substance is altered during transport across the
membrane

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 Cytoplasm

• Cytoplasm is the substance inside the plasma

membrane
• Contains 80% water, proteins, carbohydrates, lipid,
inorganic ions, low-MW compounds
• Major structures: nuclear area (DNA), ribosomes,
inclusions
• Protein filaments are most likely responsible for the rod
and helical shapes of bacteria
• Prokaryotes cytoplasm lack of cytoskeleton and
cytoplasmic streaming

Nuclear area (nucleoid)

• Bacterial chromosomes: single long, continuous,

circularly, double-stranded DNA
• Bacterial chromosomes aggregate to form a visible
mass called nucleoid
• Genetic information for cell’s structure and function
• Not surrounded by nuclear envelope, lack histones
• The chromosomes is attached to plasma membrane
• Protein in plasma membrane are believed to be
responsible for replication of the DNA and segregation
of the new chromosomes daughter cells during division

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 Nuclear area (plasmids)

• Plasmids: circular, double-stranded DNA molecules

• Extrachromosomal genetic elements
• Contain 5-100 genes and not crucial for its life
• Plasmids can be gained or lost without harming the cell
• Carry genes for antibiotic resistance, tolerance to toxic
metals, the production of toxins and synthesis of
enzymes
• Plasmids responsible for conjugation and transmissible
between cells during conjugation
• Plasmids can be transferred between species or genus
• Plasmids DNA is used for gene manipulation

Plasmids (R factor)

• RTF: plasmid replication and transfer by conjugation

• R determinant: production of enzymes (exoenzymes,
exotoxins), adhesins, inactivate drugs

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 Ribosomes

• Synthesis protein
• Two subunits: protein (40%) and rRNA (60%)
• Smaller and less dense compared to eukaryotes cells
• 70S: 30S (1 mol. rRNA) and 50S (2 mol. rRNA)
• Medically targeted: aminoglycosides (30S), erythromycin
& chloramphenicol (50S)

Inclusions

• Reserve deposit
• Evidence suggests that macromolecule concentrated
in inclusions avoid the increase in osmotic pressure
that would result if the molecule were dispersed in the
cytoplasm
• Serve as a basis of identification
• Metachromatic granules (volutin): inorganic phosphate
(polyphosphate), synthesis ATP
• Polysaccharide granules: glycogen and starch
• Lipid inclusions: polymer poly-β-hydroxybutyric acid
• Sulfur granules: oxidizing sulfur and sulfur-containing
compound

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 Inclusions

• Carboxysomes: contain enzymes ribulose 1,5-

diphosphate carboxylase for CO2 fixation
• Gas vacuoles: gas vesicle, hollow cylinders covered by
protein, maintain buoyancy so the cells can remain at
the depth in the water appropriate for them to receive
sufficient amount of O2, light and nutrients
• Magnetosomes: Fe3O4, move downward until reach a
suitable attachment site, may protect the cell against
H2O2 accumulation

Endospores
• Differentiated cells, highly resistant
• Ideal structure for dispersal (wind, water & animal gut)
• 20 genera (Bacillus, Clostridium, Sporosarcina and
Heliobacterium)
• Dipicolinic acid (DPA), located in the core
• Rich of Ca2+, combined with DPA, 10% of dry weight
• Reduce water availability, helping to dehydrate
• The complex intercalates (insert between bases) in
DNA → stabilize it to heat denaturation
• SASPs (small acid-soluble proteins)
• Bind tightly to DNA in the core and protect it
• Carbon and energy source during germination

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 Endospores

Endospores

• Exosporium: protein
• Spore coat: spore-
specific protein
• Cortex: loosely cross-
linked peptidoglycan
• Core or spore protoplast:
core wall, cytoplasmic
membrane, cytoplasm,
nucleoid, ribosomes,
other cellular essentials

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 Endospores

Bacillus subtilis, 8 hours for the sporulation process

Endospores germination

• Activation
• Heating several minutes at sublethal temperature
• Germination
• Placed in the specific nutrients (AAs, alanine)
• Rapid process (several minutes)
• Loss microscopic refractility, loss of resistance to
heat and chemical, loss of Ca2+-DPA and cortex,
SASPs are degraded
• Outgrowth
• Visible swelling due to water uptake
• Synthesis of new RNA, proteins and DNA

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