PLASMA MEMBRANE:

The cytoplasmic membrane act as a hydrophobic barrier for the penetration of most water soluble molecules. However specific

proteins in the membrane facilitate the passage of small molecules (nutrients and waste products) across the membrane.
The cytoplasmic membrane also contains various enzymes involved in respiratory metabolism and in synthesis of capsular
and cell wall components.

The Plasma (Cytoplasmic) Membrane

The plasma membrane of prokaryotes consists primarily of phospholipids which are the most
abundant chemicals in the membrane, and proteins. Immediate below the cell wall is cytoplasmic membrane
which is similar in both gram + ve and gram -ve bacteria.
It is about 75 nm thick bilayered membrane and is composed primarily of phospholipids (about 20-30%) and proteins (about 6070%).
Each phospholipid molecule contains a polar head, composed of a phosphate group and glycerol
that is hydrophilic (water-loving) and soluble in water, and nonpolar tails, composed of fatty acids
that are hydrophobic (water-fearing) and insoluble in water (Figure 4.14c). The polar heads are on
the two surfaces of the lipid bilayer, and the nonpolar tails are in the interior of the bilayer.

The phospholipid form a bilayer in which most of the proteins are tenaciously held (integral proteins). These protein only
can be removed by destruction of the membrane or by the treatment with detergents. Other proteins are loosely attached (peripheral
proteins) and can be removed by mild treatment such as osmotic shock. Each phospholipids molecule of the bilayer has both
hydrophilic head facing outwards and a hydrophobic tail facing towards each other. The lipid matrix of the membrane has fluidity.
This type of structure of plasma membrane is known as fluid mosaic model. (OR)

The protein molecules in the membrane can be arranged in a variety of ways. Some, called
peripheral proteins, are easily removed from the membrane by mild treatments and lie at as
enzymes that catalyse chemical reactions, as a scaffold for support, and as mediators of changes
in membrane shape during movement. Other proteins, called integral proteins, can be removed
from the membrane only after disrupting the lipid bilayer (by using detergents, for example). Most
integral proteins penetrate the membrane completely and are called transmembrane proteins.
Some integral proteins are channels that have a pore, or hole, through which substances enter and
exit the cell.
Many of the proteins and some of the lipids on the outer surface of the plasma membrane
have carbohydrates attached to them. Proteins attached to carbohydrates are called
glycoproteins; lipids attached to carbohydrates are called glycolipids. Both glycoproteins and
glycolipids help protect and lubricate the cell and are involved in cell-to-cell interactions. For
example, glycoproteins play a role in certain infectious diseases. The influenza virus and the toxins
that cause cholera and botulism enter their target cells by first binding to glycoproteins on their
plasma membranes.

Protein-to-lipid ratios of bacterial plasma membranes are approximately 3: I, close to those for
mitochondrial membranes.
Unlike eukaryotic cell membranes, the bacterial membrane has no sterols, and bacteria lack the enzymes required
for sterol biosynthesis. One exception is the wall-less prokaryote Mycoplasma, which contains
membrane sterols.

Although their composition is similar to that of inner membranes of Gram-negative species, cytoplasmic
membranes from Gram-positive bacteria possess a class of macromolecules not present in the Gram-negative
membranes.

Functions

The most important function of the plasma membrane is to serve as a selective barrier through
which materials enter and exit the cell. In this function, plasma membranes have selective
permeability (sometimes called semipermeability).
The permeability of the membrane depends on several factors. Large molecules (such as proteins)
cannot pass through the plasma membrane, possibly because these molecules are larger than the
pores in integral proteins that function as channels.
But smaller molecules (such as water, oxygen, carbon dioxide, and some simple sugars) usually
pass through easily. Ions penetrate the membrane very slowly.
Substances that dissolve easily in lipids (such as oxygen, carbon dioxide, and nonpolar organic
molecules) enter and exit more easily than other substances because the membrane consists
mostly of phospholipids. The movement of materials across plasma membranes also depends on
transporter molecules, which will be described shortly.
Plasma membranes are also important to the breakdown of nutrients and the production of energy.
The plasma membranes of bacteria contain enzymes capable of catalysing the chemical reactions
that break down nutrients and produce ATP.

Cytoplasm
Cytoplasm is about 80% water and contains primarily proteins (enzymes), carbohydrates, lipids,
inorganic ions, and many low molecular- weight compounds. Inorganic ions are present in much
higher concentrations in cytoplasm than in most media.
Cytoplasm is thick, aqueous, semi-transparent, and elastic. The major structures in the cytoplasm of
prokaryotes are a nucleoid (containing DNA), particles called ribosomes, and reserve deposits called
inclusions. Protein filaments in the cytoplasm are most likely responsible for the rod and helical cell
shapes of bacteria. Prokaryotic cytoplasm lacks certain features of eukaryotic cytoplasm, such as a
cytoskeleton and cytoplasmic streaming.

NUCLEAR MATERIAL:
The nucleoid of a bacterial cell usually contains a single long, continuous, and frequently
circularly arranged thread of double-stranded DNA called the bacterial chromosome.

Bacterial chromosomes are not surrounded by a nuclear envelope (membrane) and do not
include histones. The nucleoid can be spherical, elongated, or dumbbell shaped.

In actively growing bacteria, as much as 20% of the cell volume is occupied by DNA because
such cells presynthesize nuclear material for future cells. The chromosome is attached to the
plasma membrane. Proteins in the plasma membrane are believed to be responsible for replication
of the DNA and segregation of the new chromosomes to daughter cells during cell division.

Bacteria lack a well-defined nucleus. Its genetic material is designated under the area near the centre of the cell and
regarded as nucleoid/ chromatin body or bacterial chromosome since it consist of single circular DNA molecule in which all
genes are linked. It can be made visible under the light microscope by Feulgen staining which is specific for DNA.

Under Electron microscopy it appears as a light area with a delicate fibrillar structure. Its size measures, about 1000~
in length and 3 nm in diameter. Its molecular weight is nearly 3-5 x 10 9

It has about 4000 genome whose replication is by semiconservative method. The bacterial chromosome differ from

eukaryote chromosome in lacking histone (basic) protein however polyamines may be found to some of the phosphate group
of the bacterial DNA. Polyamines are small molecules rich in amino groups.

The cross section of nucleoid show 500-900 strands folded back and forth several hundred times. The size of
nucleoid increases during replication.

Bacterial chromatin does not contain basic histone proteins, but low-molecular-weight polyamines and
magnesium ions may fulfil a function similar to that of eukaryotic histones. Despite the differences between
prokaryotic and eukaryotic DNA, prokaryotic DNA from cells infected with bacteriophage g, when visualised by
electron microscopy, has a beaded, condensed appearance not unlike that of eukaryotic chromatin.

Plasmid and Episomes: In many bacteria in addition to nucleoid there is an extra small circular DNA segment
which is in the form of ring is called Plasmid. There replication is autonomous. This extrachromosomal DNA fragment
was first discovered by Lederberg (1952). They are extrachromosomal, self-replicating and stably inherited, whose size
ranges from about 20-100 kb pairs (a bacterial chromosome is about 4000 kbp). Plasmid has an independent replication
and contain own system for initiating and controlling the replication. The numbers of genes in plasmid vary from 3-4
who has no role in viability and growth of bacteria.
Under certain conditions, however, plasmids are an advantage to cells. Plasmids may carry genes
for such activities as antibiotic resistance, tolerance to toxic metals, the production of toxins, and
the synthesis of enzymes. Plasmids can be transferred from one bacterium to another. In fact,
plasmid
Two types of plasmids have been identified:
1. Conjugative Plasmid : It carries genes that promote the transfer of plasmids from host cell to a recipient cell by
conjugation.
2. Non-conjugative Plasmid: It can't promote its own transfer by conjugation.

Episomes are the plasmid which get integrated into the bacterial chromosomes. It was discovered by Jacob, Schaeffer
and Wollman (1960). The first mentioned plasmid responsible for fertility was named as fertility factor or F factor. It
plays an important role in conjugation in E. coli. It is about 94.5 kb long carries the gene responsible for cell attachment
and plasmid transfer between specific strains of bacteria during conjugation.
Important characteristic of naturally occurring plasmids:
1. They replicate independently of the main chromosome.
2. They are species specific to one or few species of bacteria.
3. They can undergo reversible integration into bacterial chromosome.
4. A few plasmid can pick up and transfer chromosomal gene.
5. They can be transferred by conjugation.
6. They usually contain up to 40 genes.
7. They do not occur free in nature.

The prokaryotic nucleoid the equivalent of the eukaryotic nucleusis structurally simpler than the true eukaryotic
nucleus, which has a complex mitotic apparatus and surrounding nuclear membrane.

Inclusions
Within the cytoplasm of prokaryotic cells are several kinds of reserve deposits, known as
inclusions. Cells may accumulate certain nutrients when they are plentiful and use them when the
environment is deficient. Evidence suggests that macromolecules concentrated in inclusions avoid
the increase in osmotic pressure that would result if the molecules were dispersed in the cytoplasm.

Some inclusions are common to a wide variety of bacteria, whereas others are limited to a small
number of species and therefore serve as a basis for identification.
Metachromatic Granules

Metachromatic granules are large inclusions that take their name from the fact that they
sometimes stain red with certain blue dyes such as methylene blue. Collectively they are known as
volutin. Volutin represents a reserve of inorganic phosphate (polyphosphate) that can be used in
the synthesis of ATP. It is generally formed by cells that grow in phosphate-rich environments.
Metachromatic granules are found in algae, fungi, and protozoa, as well as in bacteria. These
granules are characteristic of Corynebacterium diphtheria, the causative agent of diphtheria; thus,
they have diagnostic significance.
Polysaccharide Granules

Inclusions known as polysaccharide granules typically consist of glycogen and starch, and their
presence can be demonstrated when iodine is applied to the cells. In the presence of iodine,
glycogen granules appear reddish brown and starch granules appear blue.
Lipid Inclusions

Lipid inclusions appear in various species of Mycobacterium, Bacillus, Azotobacter, Spirillum, and
other genera. A common lipid-storage material, one unique to bacteria, is the polymer poly-hydroxybutyric acid. Lipid inclusions are revealed by staining cells with fat-soluble dyes, such as
Sudan dyes.
Sulphur Granules

Certain bacteria e.g.the sulphur bacteria genus Thiobacillusderive energy by oxidizing sulphur
and sulphur-containing compounds. These bacteria may deposit sulphur granules in the cell,
where they serve as an energy reserve.
Carboxysomes

Carboxysomes are inclusions that contain the enzyme ribulose-1,5-diphosphate carboxylase.

Photosynthetic bacteria use carbon dioxide as their sole source of carbon and require this enzyme
for carbon dioxide fixation. Among the bacteria containing carboxysomes are nitrifying bacteria,
cyanobacteria, and thiobacilli.
Gas Vacuoles

Hollow cavities found in many aquatic prokaryotes, including cyanobacteria, anoxygenic

photosynthetic bacteria, and halobacteria are called gas vacuoles. Each vacuole consists of rows
of several individual gas vesicles, which are hollow cylinders covered by protein. Gas vacuoles
maintain buoyancy so that the cells can remain at the depth in the water appropriate for them to
receive sufficient amounts of oxygen, light, and nutrients. Some bacteria living in aquatic habitat form gas
vacuoles that provide buoyancy. In light microscope these are bright, retractile bodies and can be made to collapse under
pressure and there by lose their refraction. The wall is made up of protein ego Non-pigmented members of phototroph
bacteria like Polynema, Holobacterium & Clostridium.
Magnetosomes

Magnetosomes are inclusions of iron oxide (Fe3O4) surrounded by invaginations of the plasma
membrane. Magnetosomes are formed by several gram-negative bacteria such as Magnetospirillum
magnetotacticum and act like magnets (Figure 4.20). Bacteria may use magnetosomes to move
downward until they reach a suitable attachment site. In vitro, magnetosomes can decompose
hydrogen peroxide, which forms in cells in the presence of oxygen. Researchers speculate that
magnetosomes may protect the cell against hydrogen peroxide accumulation.
CYTOPLASMIC INCLUSIONS
Concentrated deposits of a variety of reserve materials are detectable in the cytoplasm of some bacteria. They have high
molecular weight and usually osmotically inert. They are mainly of three types:

(i) Polymetaphosphate (Po3-)n : They are also known as vo0lutin granule or metachromatic granules which appear reddish
violet after staining with methylene blue. They are most common in Spirillum volutans, Conjnebacterium diphtheriae and
Mycobacteria. This is required during nucleic acid synthesis.
(ii) Poly p-Hydroxybutyrate : They serve as source of metabolic energy and stained with sudan black. It is found in Bacillus
megatherium who contain about 60% of the dry weight.
(iii) Polyglucan Granules: They appear blue, reddish blue or brown when stained with iodine.
(iv)Sulphur inclusions: They are found in bacteria growing in environment rich in sulphur e.g. purple sulphur bacteria which
utilize H2S either as e- donor during photosynthesis or non-photosynthetic bacteria e.g. Beggiatoa and Thiothrix.

Ribosomes
All eukaryotic and prokaryotic cells contain ribosomes, which function as the sites of protein
synthesis.
cytoplasm of a prokaryotic cell contains tens of thousands of these very small structures, which give
the cytoplasm a granular appearance (see Figure 4.6).
Ribosomes are composed of two subunits, each of which consists of protein and a type of RNA
called ribosomal RNA (rRNA).Prokaryotic ribosomes differ from eukaryotic ribosomes in the number
of proteins and rRNA molecules they contain; they are also somewhat smaller and less dense than
ribosomes of eukaryotic cells.
Accordingly, prokaryotic ribosomes are called 70S ribosomes (Figure 4.19), and those of eukaryotic
cells are known as 80S ribosomes. The letter S refers to Svedberg units, which indicate the relative
rate of sedimentation during ultra-high-speed centrifugation. Sedimentation rate is a function of the
size, weight, and shape of a particle. The subunits of a 70S ribosome are a small 30S subunit
containing one molecule of rRNA and a larger 50S subunit containing two molecules of rRNA.
Several antibiotics work by inhibiting protein synthesis on prokaryotic ribosomes. Antibiotics such as
streptomycin and gentamicin attach to the 30S subunit and interfere with protein synthesis. Other
antibiotics, such as erythromycin and chloramphenicol, interfere with protein synthesis by attaching
to the 50S subunit. Because of differences in prokaryotic and eukaryotic ribosomes, the microbial
cell can be killed by the antibiotic while the eukaryotic host cell remains unaffected.
Ribosomes: Ribosomes are found in free floating conditions and are randomly distributed in the cytoplasm. They constitute
about 30% of the total weight of the bacterium (10,000 -15000 ribosome in a bacterial cell). During protein synthesis a
number of ribosomes are held together by mRNA and form polyribosomes. The number of ribosome is directly proportional
to the rate of protein synthesis.
The ribosome of prokaryote are of 70s type whose molecular weight is about 2.7 million. Each 70s ribosome is composed of
two subunits larger 50S and a smaller 30S. At low concentration of Mg+2 ions these 70S ribosome is dissociated into its two
subunits.
Ribosome of E. coli bacteria is made up of 63% RNA and 37% protein or they are in 2:1 ratio. Many antibiotics like
streptomycin and tetracycline who inhibit the protein synthesis in bacterial cell have a main target on the ribosome of the
bacterium.
Lamellae or Chromatophore : Lamellar thylakoid or vesicles are found in many photosynthetic bacteria. They are known as
chromatophores. Lamellae are synthesized by two unit membranes. These membranes are distributed throughout the
cytoplasm. These chromatophore are hollow rounded structure with a diameter of about 300A 0. They bears photosynthetic
pigments, enzyme required for light reaction, ETS system of photophosphorylation. They lack the enzyme required for dark
reaction.
Mesosome : Bacterial cell do not contain membrane bound organelles (Viz. mitochondria, chloroplast, golgi apparatus etc).
But in bacteria the cytoplasmic membrane have specialized invagiantions that can increase their surface area for certain

function.
Especially in gram positive bacteria these membrane invaginations are in the form of convoluted tubules and vesicles termed
mesomes. They are well developed in bacilli, and may be 2-4 in number in each cell. Their number is higher in those bacteria
involved in higher respiratory activity e.g. Azatobacter. The mesosomes may be central or peripheral in position. The central
mesosome penetrate deeply into the cytoplasm and located near the middle of the cell, and seemed to be attached to the
genetic material of the cell and thought to be involved in replication of DNA and cell division. While the peripheral
mesosome show only a shallow penetration into the cytoplasm seem to be involved in export of exocellular enzymes such as
penicillinase. The nature of mesosome was initially considered to be equivalent to mitochondria of higher plants and
considered to be pockets of respiratory activity since they lack outer membrane they are not considered analogous to
mitochondria. Along with this it is also devoid of many plasma membrane enzymes viz. ATPase, dehydrogenase, and
cytochrome. It has been suggested that mesosomes help in the formation of septum.23x