Bacte Lecture 2

BACTERIAL CELL MORPHOLOGY AND

COMPOSITION
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TWO STRUCTURAL TYPES OF CELLS

 Cell = basic unit of life

o Prokaryotic: ALL BACTERIA; nucleus is absent, instead there is NUCLEOID
 E.g. (In microorganisms) Bacteria, blue green algae, mycoplasmas,
spirochetes
o Eukaryotic: PLANTS AND ANIMALS; membrane-bound organelles + nucleus
 E.g. (In microorganisms) Fungi, Protozoa, Algae
COMPARISON OF PROKARYOTES AND EUKARYOTES
Nucleus: Absent in prokaryotes. Present with nuclear membrane in eukaryotes.
Organelles: Absent in prokaryotes. Present in a variety of forms in eukaryotes.
DNA Structure: Single closed loop with almost naked strand and very little protein in
prokaryotes. Multiple chromosomes in nucleus with structural protein associated with DNA in
eukaryotes.
Chlorophyll: When present, dissolved in cytoplasmic membranes in prokaryotes. When present,
dissolved in chloroplast membranes in eukaryotes.
Ribosomes: In prokaryotes, smaller than eukaryotic ribosomes and are free in cytoplasm. In
eukaryotes, larger than prokaryotic ribosomes and are free or bound to membranes.
Cell walls: Generally present in prokaryotes with complex chemical composition. Present in
some types of eukaryotes with complex chemical composition.
Flagella: Follows a rotating movement in prokaryotes. Follows a whipping movement in
eukaryotes.
Cilia: Absent in prokaryotes. Present in eukaryotes.
Reproduction: In prokaryotes, by fission. In eukaryotes, by mitosis.
WHAT IS A BACTERIUM/BACTERIAL CELL?
- Unicellular
- Prokaryotic
- Contains both DNA and RNA
- Does not have histones (proteins required for mitotic division)
- Reproduces through asexual reproduction (binary fission)
BACTERIAL MORPHOLOGY:
 3 most common shapes: rod-like (bacilli), spherical (cocci), spiral (spirochete)
 Some rare shapes: star-shaped and square-shaped
 Variations in arrangement:
o “Strepto—” = chains
o “Diplo—” = in pairs
o Tetrads—” = in fours
o “Sarcinae”—packets of eight
 Size: GIANT BACTERIA = Thiomargarita (750 micrometers); nanobacteria (0.02
micrometers)
BACTERIAL CELL STRUCTURE/COMPONENT:
- Cytoplasm: other parts of the microorganisms can be found
- Nucleoid: where genetic material can be found
- Ribosomes: important for protein synthesis
- Cell wall & cytoplasmic membrane: responsible for integrity of the cell
CELL WALL
 Layer that is usually fairly rigid that lies outside the plasma membrane
 One of the most important structures because
o Confers shape
o Protects the cell from osmotic lysis (important function)
o Anchors the flagellum
o Protects the cell from toxic substances and pathogens
 o Used in identification (by studying the composition of the cell wall)
- C, U, P, P, A
 Bacteria can be divided into two big groups based on cell wall structure (Gram + and
Gram -)

COMPARISON BETWEEN GRAM + AND GRAM – CELL WALL

Gram (+) = THICK PEPTIDOGLYCAN
Gram (-) = relatively THIN PEPTIDOGLYCAN with outer membrane and periplasmic space
GRAM POSITIVE CELL WALL
Characteristics
A. Thick layer of peptidoglycan
 Peptidoglycan (PG); aka murein; mucopeptide
o Polymer of disaccharide linked by polypeptide (NAM and NAG, also with an
amino acid side chain)
o Insoluble, porous, big polymer
o > 50% of the cell wall’s dry weight
o Could be isolated as murein sacculus (in aid of diagnosis or identification of
bacteria)
 Peptidoglycan sub-unit
 o 4 amino acid components attached to NAM
 L-Alanine
 D-Glutamic acid
 D-Alanine
 Meso-diaminopimelic acid
o 2 major sugars:
 N-acetylglucosamine (NAG)
 N-Acetylmuramic acid (NAM)
o B(1,4) linkage (sensitive to lysozyme)
B. Presence of Teichoic acids
 Polymers of repeating units of glycerol or ribitol joined by
phosphates
 Amino acids (D-ala) or sugars (glu) are attached by
glycerol/ribitol
 Covalently linked to murein through muramic acid
 Connected/embedded in PG layer or to membrane/lipids
 LIPOTEICHOIC ACID
o Linear polymers of 16-40 phosphodiester-linked
glycerophosphate residues covalently linked to the
cell membrane
o Responsible for gram reaction or gram positivity of
microorganism
  Components:
o Peptidoglycan
o Teichoic acid
o Integral proteins
 Properties/Functions of Teichoic Acid
1. Highly antigenic – reacts during serological reaction; indicator for antigen-antibody
reaction
2. Anchor the wall to the cell membrane
3. Provide high density of regularly oriented charges
4. Storage of phosphorus
5. Facilitate attachment of bacteriophage (bacteriophage = virus going to infect the
bacteria; utilized in the manipulations of cells during genetic engineering)
6. Inhibit activity of autolytic enzymes, which hydrolize the murein (autolytic enzymes
= when activated, self-destruction occurs to cell wall, and ultimately, death of
microorganism)
- HAPSFI
 Other substances may be found in the cell wall:
o Teichuronic acid – acidic polysaccharides containing uronic acids
o Neutral polysaccharides – important in classification of some gram-positive
bacteria
o Other glycolipids – may substitute for whatever function of the LTA
o Mycolic acids – waxy lipids found in Mycobacterium (significant component;
responsible for the acid-fastness of some organisms; BASIC FUCHSIN = dye
utilized in acid-fast staining and has strong affinity to the mycolic acid)
  PERIPLASMIC SPACE
o Gram (+): gap between PM and CW
o Gram (-): gap between PM and OM

GRAM NEGATIVE CELL WALL

1. Peptidoglycan
 Thin: 1-2 layers in E. coli
 Constitute <5-10% of a wall’s dry weight (vs Gram +’s >50%)
 May be more of a gel than a compact layer
2. Outer membrane
- Compensates to Gram (-)’s thin layer of peptidoglycan; quite complex
 Located above/external to peptidoglycan layer
 Like the cytoplasmic membrane
 Other main components:
o Lipopolysaccharides (LPS) (consists of several fatty acids in the form of the
circular figures + sugar in the form of hexagonal shapes)
o Lipoproteins
o Outer membrane proteins (porins)
LIPOPOLYSACCHARIDES
 Lipids and carbohydrates
 Outer layer of the outer membrane
 Endotoxin
 Consists of three parts (responsible or components of outer membrane)
o Lipid A
o Core Polysaccharide
o O-specific polysaccharide (Outer-specific polysaccharide/O-Antigen/Outer
membrane layer)

LIPID A

 Embedded in the membrane as part of the lipid bilayer

 Hydrophobic
 Composed of 2 glucosamine (sugar) residues (N-acetylglucosamine) linked with B-1,6
(backbone) with four identical fatty acids (specific type is dependent on species of
bacteria)
- In E. coli and Salmonella typhimurium: fatty acids are B-hydroxymyristic acid (or 3-
hydroxytetradecanoic acid); 2 of the fatty acids are esterified with long chain saturated
fatty acids
CORE REGION

 Consists of an outer and inner core

 OUTER CORE
o Shows high to moderate variability
o Consists of hexoses
 INNER CORE
o Shows low structural variability
o Consists 2-keto-3-deoxyoctanate (KDO), heptose, ethonalamine (attached to
heptulose), and phosphate
O-ANTIGEN
- Outermost layer; responsible for antigenicity of a particular bacteria
- Short polysaccharide extending outward from the core
- Consists of peculiar sugars which varies between bacterial strains
- Not essential for viability (viability = ability to work successfully)

SUMMARY
 Lipid A
 Core polysaccharide
  O (Outer) side chain = consists of several sugars; composition depends on species;
variation in sugar means a specific species is being described
IMPORTANCE OF LPS
 Avoidance of host defenses (O-antigen)
 Contribution to the negative charge on the cell’s surface
 Stability of membrane structure
 Acts as endotoxin (endotoxin = toxic heat-stable lipopolysaccharide substance present in
the outer membrane of gram-negative bacteria that is released from the cell upon lysis)
LIPOPROTEINS (BRAUN’S LIPOPROTEIN)
 Mediate interconnection between the OM and murein
 Synthesized within the cell and contains a leader sequence of ~20 amino acids at its
amino terminal end
 After transport through the CM, the leader sequence is removed, and the terminal residue
is modified to make it hydrophobic (facilitates integration into the OM)
PORINS
 Small holes that will pass through the cell wall, to the cellular membrane, to the cell; not
seen by the naked eye because microscopic; facilitates diffusion, ergo there is no need for
transport proteins (direct from outside to inside of the cell—more about transport
mechanisms in Bacterial Physiology topic)
 Form small hydrophilic channels through the outer envelope allowing the diffusion of
neutral and charged solutes of MW <600 daltons
 Three identical units
 Associate to form membrane holes
 Transmembrane
- Though pictures seem to show porins stop until periplasmic space/peptidoglycan only?
IMPORTANCE OF THE OM
 Proteins in OM are used as attachment sites by bacteriophages (virus, during invasion,
anchors to OM)
 Permeability barrier to heavy metals, lipid-disrupting agents, and larger molecules
(maintain integrity of the cell)
 Outer surface with strong negative charge is important in evading phagocytosis
 Highly variable O-antigen reduces complement binding (same C’ from IS)
 O-antigen provides host with multiple antigenic structures in various stains (antigenicity
of bacteria)
 LPS complex is a bacterial endotoxin causing a variety of pathophysiological reactions
ranging from fever to death
 May be involved in adherence
- APERMBA
PERIPLASM
 A separate compartment between the cell membrane and outer membrane in Gram (-)
bacteria
 Seen in electron micrographs as space but is considered an aqueous compartment
 Activities:
o Redox reactions
o Osmotic regulation
o Solute transport
o Protein secretion
o Hydrolysis
- HORPS
PERIPLASM: COMPONENTS AND FUNCTIONS
1. Oligosaccharides – thought to be involved in the osmotic regulation of the periplasm
because their amounts decrease when the cells are grown in media of high osmolarity
- Osmolarity = concentration of dissolved particles of chemicals and minerals; higher
osmolarity = more particles in media
2. Solute binding proteins – bind to solutes and deliver solutes to specific transporters in the
membrane
3. Cytochrome – cyt c
- Part of Electon Transport Chain, which can be found in the cytoplasmic membrane and
periplasmic area
4. Hydrolytic enzymes – degrade nutrients to smaller molecules that can be transported
across the membrane by specific transported
a. Detoxifying agents – e.g. B-lactamase (enzymes that attack beta-lactam
antibiotics)
 b. TonB protein – required for the uptake of several solutes (iron siderphores, vit
B12) that do not diffuse through the porin (e.g. charged particles from outside
membrane to inside the cell)
- COSH
PLASMA MEMBRANE

 Most dynamic structure of a prokaryotic cell

 Definitive structure of the cell
 Has a variety of functions in the energy generation and biosynthesis
- Since prokaryotes have no mitochondria
 Contains transport proteins, binding proteins, sensing proteins, and enzymes involved in
metabolic processes
- Will be discussed in Bacterial Metabolism
 Absolute requirement of all living organisms (responsible for the semi-permeability and
integrity of a particular cell)
 Chief point of contact with the environment
PLASMA MEMBRANE: FUNCTIONS
1. Transport of nutrients (active and passive transport—will be discussed later on)
2. Location of a variety of crucial metabolic processes
o Synthesis of membrane lipids
o Wall murein synthesis (peptidoglycan)
o Assembly and secretion of extracytoplasmic proteins
o Respiratory electron transport
- WARS
3. Chromosome segregation
4. Establishment of electrochemical gradient
5. Motility (because flagella is anchored to inner portion of cytoplasmic membrane)
6. ATP (energy currency of bacterial cell) Synthesis
7. Intracellular signaling (Transduction mechanisms—bacterial communication)
8. Respond to environmental signals (responsible in sensing chemotaxins and
antibiotics/stimulus then come up with the movement of the flagella)
- TLECMAIR
PLASMA MEMBRANE: 3 IMPORTANT FUNCTIONS
1. Permeability Barrier – prevents leakage and functions as a gateway for transport of
nutrients into and out of cell
- loss of this function leads to cell death; plasmatisis (?) or bursting of bacterial cell due to
uncontrolled osmotic mechanism
2. Protein Anchor – site of many proteins involved in transport, bioenergetics, and
chemotaxis
- So many proteins embedded on cell membrane with many responsibilities (e.g. transport,
integral)
3. Energy Conservation – site of generation and use of the proton motive force
- PEA

COMPOSITION
 Phospholipids
- Bilayer—in eukaryotes, consists of phosphatidylcholine and phosphatidylethanolamine—
responsible for semi-permeability
 Peripheral proteins
 Integral proteins
1. PHOSPHOLIPIDS
 Most membrane associated lipids are structurally asymmetric
- Glycerol: backbone of phospholipids; 2 esterified fatty acids; phosphate group in carbon
#3

 For every 1 g of cells: 91 mg phospholipid = 129 micromole phosphatidylethanolamine

where 1 mole: 750 micrograms PE
o Phosphatidylethanolamine = major phospholipid in the cytoplasmic membrane;
formed from glycerol-3-P, serine, and fatty acids in the ratio of 1:1:2
o Phosphatidylglycerol = next most prevalent; formed from glycerol-3-P and fatty
acids in equimolar rations
o Fatty acids – varies depending on the species and environmental conditions
- (not going to focus on this information)
2. PROTEINS
 Two types of membrane proteins have been identified
a. Integral proteins – embedded in the CM; amphipathic (both hydrophobic and
hydrophilic parts); bound to the fatty acids of the phospholipids via
hydrophobic bonding
b. Peripheral proteins – attached to the membrane surfaces by ionic interactions
 - Sometimes some substances will not be allowed to enter the cell due to incompatibility,
in terms of charges, with the peripheral proteins
 Most are lipoproteins which contain a lipid tail on the amino terminus of the protein that
anchors the protein in the membrane
3. MEMBRANE STEROIDS AND HOPANOIDS
 Rigid, planar molecules found associated with bacterial CM
 Molecules similar to sterol
- Similar in function with the cholesterol—responsible for rigidity, which are embedded on
the bilipid layer

FLUID MOSAIC MODEL

 Most widely accepted model of the CM
 Shows that the CM is a lipid bilayer with which proteins and lipids “float” freely
- Proteins and other substances in lipid bilayer allow floating movement—not too rigid, not
too prone to break
- Once bilipid layer is destroyed, it will lose its integrity, and will lead to cell death or cell
lysis
THE CYTOPLASM
 Consists of aqueous solution of three groups of molecules
o Macromolecules (proteins, messenger RNA, transport RNA, etc.)
o Small molecules which serve as energy sources, precursors of macromolecules,
metabolites, or vitamins
o Various organic and inorganic ions and cofactors
- Literally cytoplasmic components + nuclear materials can be found inside the cytoplasm
 Structural components:
o Nucleoid
 RNA, DNA, proteins
o Ribosomes
 Important in protein synthesis
o Inclusion bodies
 Serves several functions depending on the type and species of bacteria
 - 3 important and prominent components that can be observed inside the cytoplasm
PROKARYOTIC CYTOSKELETON
 Cytoskeletal filaments that are structurally similar to their eukaryotic counterparts
o Cresentin (intermediate filament proteins) – responsible for cell shape
 One of the very important
o FtsZ (tubulin) – cell dvision
 During binary fission
o MreB (actin) – cell shape
- If these particular genes are being mutated, there must be a change in bacterial
shape/bacterial components/arrangement of bacteria
MESOSOMES
- Play a very important role
 Invaginations of the plasma membrane in the shape of vesicles, tubules, or lamellae
 Believed to be involved in cell septum formation (cell septum formation = initial step in
cellular division/binary fission), electron transport, photosynthesis, cell wall formation,
DNA replication, segregation, and sporulation (initial steps as well)
INCLUSION BODIES
 Distinct bodies that may occupy a substantial part of the cytoplasm
 May be organic or inorganic
 Some inclusion bodies lie free in the cytoplasm or enclosed by a shell consisting of
proteins or a membranous structure composed of proteins and phospholipids
 Usually used for storage
- Microorganisms or bacteria may not have all of these inclusion bodies; presence is also
distinctive to the particular species of bacteria; can be utilized in the identification of the
specific species of bacteria
GRANULES AND GLOBULES
a. Glycogen – a polymer of glucose units composed of long chains formed by
a(14) glycosidic bonds and branching chains connected to them by a(16)
glycosidic bonds (1 out of 2 major polysaccharides, other one is starch)
b. Poly-ß-hydroxybutyric acid (PHB) –
contains ß-hydroxybutyrate molecules joined
by ester bonds (aka Poly-ß-hydroxyalkanoids;
can be found in a certain group of
microorganisms; serves as food source in time
of scarcity)
c. Cyanophycin granules – composed of large polypeptides
containing approximately equal amounts of amino acids,
arginine, and aspartic acid (the two said amino acids are
important because from them, other amino acids can be derived,
which are important in the survival of bacteria; serves as storage
for arginine and aspartic acid)

d. Carboxysomes – polyhedral bodies

about 100 nm in diameter; contain the
enzyme ribulose-1,5-biphosphate
carboxylase (used in the carbon
fixation portion of the Calvin cycle);
may serve as site for CO2 fixation
e. Chlorosomes – ellipsoidal vesicles
where
photosynthetic pigments are located; attached to the
plasma membrane but not continuous with it
f. Polyphosphate granules (volutin or metachromatic
granules) – linear polymer of orthophosphates
joined by ester bonds; function as storage reservoir
for phosphates or as energy reserve (there is a
species of bacteria with so much polyphosphate
granules it serves as a characteristic trait of that said
species—Cornynebacterium diphtheria)
g. Sulfur granules – used to store sulfur temporarily; accumulate in the periplasmic
space of in special cytoplasmic
globules (can be found in
photosynthetic sulfur bacteria which
can be found in extreme environments
such as in mining areas; they are
responsible for the yellow/orange
color in the soil surrounding said
areas)
h. Gas vesicles – small, hollow, cylindrical structures
composed entirely of a single small protein;
impermeable to water but freely permeable to gases (in
some microorganisms, this can be utilized for
buoyancy, like if they live in the ocean and their food is
on the surface; they fill these vesicles with gas to float
and release gases when they need to go back down)
 i. Magnetosomes – intracellular chains of
magnetite (Fe3O4), greigite (Fe3S4), or pyrite
(FeS2) particles bounded by a membrane; aids
in orienting bacteria in the Earth’s magnetic
field (very important in the migration or
movement of bacteria; since they are very
small, they usually just stay in one place and
this inclusion body helps)

RIBOSOMES
 Complex structures made of both
protein and ribonucleic acid
 Present in the cytoplasmic matrix
or loosely attached to the plasma
membrane
 Site of protein synthesis (Really
important! Where the Translation process takes place)
 Composition: 50 different proteins, 3RNA (23S, 16S, and 5S—16S utilized for analysis,
23S for molecular diagnostics, 70S for the entire process of Translation in prokaryotes
but 80S in eukaryotes)
NUCLEOID
 Irregularly shaped region where the
prokaryotic chromosome is located
 Bacterial chromosome:
 o Circular in most cases
o Very tightly coiled
o Histone-like proteins generally absent
- Genetic materials found in that particular portion of the cytoplasm; all genetic
information, we can get here
COMPONENTS EXTERNAL TO THE CELL WALL
- Not all microorganisms have these. Aka ultrastructures.
 GLYCOCALYX – general term for the thick cover or layer of polymers deposited
outside the cell
o Capsule – well-organized glycocalyx; attached firmly to the cell wall; compact;
excludes particles like India ink (can be observed using special type of stain)
o Slime layer – zone of diffused, unorganized material; loose association; does not
exclude particles (usually disintegrates during staining)
- Hollow portion surrounding particular cell
o Importance:
 Exclude viruses and most hydrophobic toxic substances (protect bacterial
cells from outside environment; when capsulated, hard to treat diseases
caused by infective agents; e.g. S. pneumoniae, Krebsiella)
 Aid attachment to surfaces
 Protection from physical injury
 Provide resistance to phagocytes (e.g. macrophages, monocytes, PMNs)
 Reservoir of stored food (in extreme environment, they process their
capsule and turn to food)
 Prevent desiccation (in extreme environment, they can stay, until
environment becomes favorable)
 Confers pathogenicity
 “Cellular garbage dump”
 Antigenicity
 PILI – hair-like structures on the surfaces of prokaryotic cells; composed of protein sub-
units called “pilins”; sometimes referred to as “fimbriae”
o Model showing the probable helical arrangement of the bacterial pilus; straight
filament; 7 nm in diameter; pilin sub-unit: 17,000 daltons

o TYPES:
1. Adhesion pili
 Involved in attachment of cells to surfaces (enhance
attachment to bacteria and multiply 24 hrs after attachment;
enhances pathogenicity)
 Composed of a single protein (pilin)
 Major determinant of bacterial virulence
  Up to 1000 per cell

2. Conjugation pili
 Vehicles for transfer of genetic information between bacteria (cell mating; extension of
pili to other species—a one-sided affair, only by one who can donate; one will only
receive)
 Composed of phosphoglycoprotein
 About 1-10 per cell
PILI are synthesized in the cytoplasm and cotranslationally translocated across the membrane
- Bacteriogenetics: other methods of transferring genetic materials aside from conjugation
(normal way)—there are two others
PROKARYOTIC MOTILITY STRUCTURES
BACTERIAL FLAGELLA
 Threadlike locomotor appendages extending outward from
the plasma membrane and cell wall
 Slender, rigid structures about 20 nm across and 15-20
micrometers long
 Rotary structure driven from a motor at the base, with the
filament acting as the propeller
FLAGELLAR ULTRASTRUCTURE
 TEM studies have shown that the bacterial flagellum is
composed of three parts:
o Filament – a hollow rigid cylinder constructed of a
single protein called “flagellin”
 Helical filament; about 14 nm in
diameter; consists of thousand of copies
flagellin (40,000 daltons)
- Origin is at the cytoplasm; stimulus comes from
cytoplasm— moving towards or away from stimulus
CLASSIFICATION BASED ON FLAGELLA:
1. No flagella = atrichous
2. One flagellum = monotrichous
3. Two single flagella at opposite sides = amphitrichous
4. Bunch of flagella on one side = lophotrichous
5. All over the cell = peritrichous
- Contributes to pathogenicity
BACTERIAL ENDOSPORE
 A type of dormant cell formed by some groups of bacteria (genetically determined; not
all bacteria are capable of spore-forming; genes being inherited or passed through from
mother down to offsprings)
 Cryptobiotic
 Highly resistant to environmental stresses
 Formed by vegetative cells in response to environmental signals that indicate a limiting
factor for vegetative growth
 A mechanism of survival
 Used in classification and identification of bacteria
o Consider shape, location, and ability to swell sporangium
 Centrally-located = Bacillus subtilis
 Subterminal spore = Bacillus; Clostridium botulinum
 Terminally-located = “drumstick/rocket appearance” Clostridium tetani
- Triggered once there is scarcity for food or in extreme environment; once converted to
spores, they can survive for years
- if they are converting vegetative cells to spores, this is how they look (Right side)
- DNA at the center  core wall  cortex  exosporium
- In favorable environment, DNA replication, exosporium and spore coat will be shed off