Cytoplasm

The cytoplasm, or protoplasm, of bacterial cells is where the functions for

cell growth, metabolism, and replication are carried out.
It is a gel-like matrix composed of water(4/5 th), enzymes, nutrients, wastes,
and gases and contains cell structures such as ribosomes, a chromosome,
and plasmids.
The cell envelope encases the cytoplasm and all its components. Unlike the
eukaryotic (true) cells, bacteria do not have a membrane enclosed nucleus.
The chromosome, a single, continuous strands of DNA, is localized, but not
contained, in a region of the cell called the nucleoid.
Nucleoid
The nucleoid is a region of cytoplasm where the chromosomal DNA is
located. It is not a membrane bound nucleus, but simply an area of the
cytoplasm where the strands of DNA are found.

Ribosomes:
Consists of RNA and protein and they are abundant in cytoplasm and often
grouped in long chains called polyribosomes.
The relative sizes of ribosomes and their subunits can determined their
sedimentation rate by centrifugation or subject to a centrifugal force.
Bacterial ribosomes have sedimentation rate of 70S; their subunits have rates of
30S and 50S. Streptomycin binds 70S ribosome and stops protein synthesis but it
can not bind 80S ribosome of eukaryotes and thereby eukaryotic cell remains
unaffected.
The unit used to measure sedimentation velocity is Svedberg, larger the S value
faster will be the sedimentation velocity. Sedimentation velocity depends upon both
shape and size and hence it is not a measure of mass.
Plasmids
Plasmids, are small, extrachromosomal genetic structures carried by many strains
of bacteria. Like the chromosome, plasmids are made of a circular piece of DNA.
Plasmids have been shown to be instrumental in the transmission of special
properties, such as antibiotic drug resistance, resistance to heavy metals, and
virulence factors necessary for infection of animal or plant hosts.

 Flagella - Flagella (singular, flagellum) are hairlike structures that provide a mean of
locomotion for those bacteria who have them.
The flagella beat in a propeller-like motion to help the bacterium move toward
nutrients; away from toxic chemicals; or, in the case of the photosynthetic
cyanobacteria; toward the light. The species of bacteria, which are motile, move due to
presence of flagella.
Genetically determined as to number and location, a species with one flagellum is
called monotrichous; with two (one on each end), amphitrichous or bipolar; with a
tuft at one end called lophotrichous; and with many flagella, peritrichous.
Prokaryotes are known to exhibit a variety of types of tactic behavior, i.e., the ability
to move (swim) in response to environmental stimuli.
chemotaxis- a bacterium can sense the quality and quantity of certain chemicals in
their environment and swim towards them (if they are useful nutrients) or away from
them (if they are harmful substances).
Positive or negative chemotaxis, phototaxis, or even magnetotaxis can determine
the direction of movement which alternates between swimming and tumbling motion.

Pili or fimbriae
(singular, pilus), small hairlike projections emerging from the outside cell
surface.
These outgrowths assist the bacteria in attaching to other cells and surfaces,
such as teeth, intestines, and rocks.
Without pili, many disease-causing bacteria lose their ability to infect because
they're unable to attach to host tissue. Gonorrhea is one of the widest spread
of the reportable diseases due to attachment fimbriae.
There are some pili with a different function: they are only found on cells
which can undergo a process known as conjugation, a means of genetic
transfer among bacteria. Such pili are call sex or F pili.

Cell wall - Peptidoglycan layer

Gram Negative bacteria

Peptidoglycan: PBP activities

endopeptidase

D-Ala

DD-carboxypeptidase

Hltje 1998

Peptidoglycan-Gram Positive bacteria

Lipopolysaccharide (endotoxin) in Gramnegative bacteria

Backbone composed of alternating pyrophosphate units (POP), linked
to glucosamine (G) and it is flanked outward by repeating units of
polysaccharides (O-antigen) and inward by long chain fatty acids
(Lipid A)

Lane 1 marker
Lane 2 O-antigen positive E. coli strain
Lane 3 O-antigen negative E. coli strain

60-90% peptidoglycan and removal of cell wall makes protoplast which bursts
easily by osmotic shock.
Retention of crystal violet is directly proportional to cell wall thickness.
Due to lack of periplasmic space the digestive enzymes and toxin are not retained
and they are released into the environment.

Only 10-20% of the cell wall is peptidoglycan remainder consists of various proteins,
lipids and polysaccharides.
Toxins and digestives enzymes are retained in periplasmic space for action,
but these enzymes or toxins do not harm the organism that produced them.
After digestion of cell wall they form spheroplast (containing CM, OM)

Mycobacteria

Cell wall is thick, contains 60% lipids and very less peptidoglycan.
The lipids make acid-fast organisms impermeable to most other strains and protect
them from acids and alkalis.
The organism grow slowly because the lipids impede entry of nutrients into cell and
cells spend large amount of energy to synthesize lipids.
They can be stained by Gram staining method and stained as Gram positive.

Nuclear region or nucleoid:

Nucleoid is centrally located, contains
DNA, RNA and proteins associated with it.
Most of the bacterium contains one circular
chromosome (except V. cholerae, which
has two). Manipulation of the microbial
genome/ chromosomes is the most
important tool in any biotechnology
industry.
Internal membrane systems:
Photosynthetic and cyanobacteria contain
internal membrane system (chromometaphore),
which has membranes derived from cell
membrane (CM) and pigments (helps to
capture lights).
Nitrifying bacteria, soil organisms those convert
nitrogen compounds into forms usable by green
plants, also have internal membranes. They
house enzymes and those are used in deriving
energy from oxidation of nitrogen compounds.

Inclusion bodies:
Bacteria can have within their cytoplasm a variety of small bodies collectively
referred to as inclusion bodies. Some are called granules and other are called
vesicles.
Granules: Densely compacted substances without a membrane covering. Each
granule contains specific substances, such as glycogen (glucose polymer) and
polyphosphate (phosphate polymer, supplies energy to metabolic processes).
Sulfur bacteria contains reserve granules of sulfur.
Polyphosphate granules are called volutin or metachromic granules, because
they display metachromasia (metachromic granules exhibit different intensities of
color). These granules are depleted in starvation.
Vesicles: Certain bacteria have specialized membrane enclosed structures
called vesicles (or vacuoles). Some aquatic photosynthetic bacteria and
cyanobacteria have rigid gas-filled vacuoles and it helps in floating at a certain
level.
Some magnetotactic bacterium, eg. Aquaspirillium
magnetotacticum, stores Magnetitite (Ferric oxide).
The presence of such magnetic inclusions enables
these bacteria to responds to magnetic fields.

There are certain bacterial cells which are called

vegetative having low level of metabolizing
Endospore:
nutrients. Vegetative cells of Bacillus and
Clostridium, produce resting stages called
endospores.
A bacterium produces a single endospore, which
merely helps the survival not reproduction. (But
the fungal spores are for survival as well as
reproduction).
Sporulation or endospore formation is a mean
by which some bacteria prepare for the
possibilities for adversities in future.
Endospores form within the cells contain very less water and are highly resistant to
heat, drying, acids, bases, certain disinfectants and even radiation.
It consists of a core, surrounded by a cortex, a spore coat and in some cases
delicately thin layer called exosporium.
The core has an outer core wall, a cell membrane, nuclear region and other cell
components. They contain dipicolinic acid (Pyridine-2,6-dicarboxylic acid, a metal
chelating agent) and large amount of calcium ions.
Endospores are capable of survive a few months to several thousands of years in
adverse environment exerting heat, cold, radiation stress.
When conditions become more favorable the spores germinate or begin to develop
into functional vegetative cells.

Flagella:

Polar monotrichous
Pseudomonas

Polar amphitrichous
Spirillium

Lophotrichous
Spirillum

Peritrichous
Salmonella
SEM of peritrichous strain Proteous
Note: bacteria without flagella are called atrichous.

In Gram-negative Bacteria

The diameter of a prokaryotic flagellium is about 1/10 th of that of

eukaryotic.
It is made up of protein subunits called flagellin.
Each flagellium is attached to cell membrane with the help of proteins
other than flagellin.
The basal region has a hook like structure and a complex basal body.
The basal body consists of a central rod or shaft surrounded by a set of
rings.
Gram negative bacteria has a pair of rings embedded in the cell
membrane and another pair of rings associated with the peptidoglycan
and lipopolysaccharide layers.
Gram-positive bacteria has one ring embedded in the cell membrane
and another in the cell wall.

In Gram-positive Bacteria

BACTERIAL MOVEMENT
Bacterial movement is produced through the action of the flagella .
Bacteria move toward attractive stimuli and away from harmful
substances and waste products in the process known as chemotaxis.
Monotrichous bacteria move forward in a simple response to chemotactic
stimuli by the counterclockwise rotation of the flagellum. This forward
movement is termed the "run".
Negative chemotaxis causes clockwise rotation of the flagellum and
results in a random tumbling motion.
Peritrichous bacteria move in a similar fashion, even though the situation
is somewhat complicated by a requirement for bundling of the flagella to
produce coordinated action during counterclockwise rotation. The
"tumble" in peritrichous bacteria is the result of bundle disruption during
clockwise flagellar rotation.
Both run and tumble are generally random movement; no one direction of
movement is more likely than any other direction.

bacterial movement contd.

Runs lasts for an average of 1.0 sec, during which the bacterium swims
about 10-20 times the length of its body.
Tumbles last about 0.1 sec and no forward progress can be achieved.
Cruising speed of a bacteria is about 10 body length/second which is
almost equal to the flying speed for us.
Bacterial chemotaxis is controlled by a complex series of events beginning
with the binding of an attractant molecule to a cell surface chemoreceptor.
Chemoreceptors are often clustered at the ends of rod-shaped cells like E.
coli. Chemoreceptors do not influence flagellar motion directly, but convey
information through a phosphorylation cascade required for signaling.
Information about the environment can be translated into motion within 200
milliseconds. A return to steady-state is assured by a coordinated feedback
loop that quickly causes a reversion to original levels of protein
phosphorylation in the absence of stimuli.

Pilli or fimbrae

Conjugation pilli found in certain

group of bacteria.

Used for genetic material transfer.

The transfer process is called

conjugation.

Antibiotic resistance transfer.

Atatchment pilli, Help bacteria

adhare to the surfaces, such as cell
surfaces and interface of water and
air.

It helps in colonization and

pathogenicity.