Form and Function

2018
 If you look closer, at the surface of your skin or
inside your digestive tract, you would see that
there are actually many organisms living there, not
just our body(Yourself).
 That’s right! You are home to around 100 trillion
bacterial cells, which outnumber your own human
cells by about 10 to one.
 It means that our body is an ecosystem. This also
means that you—for some definition of the word
you—actually consist of both of the major types of
cells: prokaryotic and eukaryotic.
 Allcells fall into one of these two broad
categories. Only the single-celled organisms of
the domains Bacteria and Archaea are
classified as prokaryotes—pro means before
and kary means nucleus.
 Only two types of cells are produced by all living
organisms on earth.
 Prokaryotes (pro. or primitive nucleus) do not have
a membrane bound nucleus
 eubacteria (true bacteria)
 archaebacteria (ancient bacteria)
 Eukaryotes (eu, or true nucleus) have a membrane
bound nucleus
 Algae
 fungi
 protozoa
 plants
 animals
 Water - 70%
 Dry weight - 30% composed of:
 DNA - 5% MW 2,000,000,000
 RNA - 12%
 protein- 70% found in:
 Ribosomes(10,000) – RNA
 Protein particles - MW 3,000,000
 Enzymes
 Surface structures
 polysaccharides - 5%
 lipids - 6%
 phospholipids - 4%
1. Appendages- flagella, pili, fimbrae
2. Cell envelope- glycocalyx, cell wall , cell membrane
3. Cytoplasm- ribosomes, granules,
nucleoid/chromosome.
ESSENTIAL STUCTURE : NONESSENTIAL STUCTURE :

1) cell wall 1) capsule

2) cell membrane 2) flagella
3) ribosomes 3) fimbriae
4) cytoplasm 4) spore
5) nucleoid 5) intracytoplasmic inclusions
6) mesosomes 6) plasmides
Peptidoglycan (polysaccharides + protein),

 Support and shape of a bacterial cell.

The three primary shapes in bacteria are:
 coccus (spherical),

 bacillus (rod-shaped)

 spirillum (spiral).

 Mycoplasma are bacteria that have no cell wall and

therefore have no definite shape.

 peptidoglycan (polysaccharides + protein)
Components of the peptidoglycan layer:
 Repeating glycan chains (N acetyl
glucosamine and N acetyl muramic acid)
 a set of identical tetrapeptide side
chains attached to N-
acetylmuramic acid
 a set of identical peptide cross
bridges
 Basis of Gram Stain Reaction
 Hans Christian Gram- 1884
 Differential Stain
 Gram Positive vs Gram Negative Cells
 Gram Positive Cells-
 Thick peptidoglycan layer with embedded
teichoic acids
 Gram Negative Cells-
 Thin peptidoglycan layer, outer membrane of
lipopolysaccharide.
 Hans Christian Gram- 1880s
 Divides bacteria into 2 main groups-
 Gram positive
 Gram negative
 Also- gram variable
 Gram nonreactive
 Gram positive bacteria
 many layers of peptidoglycan and teichoic acids.
 form a crystal violet-iodine-teichoic acid complex
 Large complex, difficult to decolorize
 The periplasmic space is the region
between outer membrane ant inner
cytoplsmic membrane.
 It contains peptidoglycan and enzyme
proteins involved in transport, digestion of
nutrients, protection against toxic substances
(eg beta-lactamases that destroy beta-
lactam antibiotics).
 It gives the shape of bacteria
 It is an osmotic and mechanical barrier
 It is a selective permeability barrier
 It has antigenic function (Ag O and R)
 It participates in processes of growth and cell division
 It has a receptor function
 Cell wall components (lipid A, lipoic acids, fragments of
PG) contribute to cytokine production and complement
activation by alternating with septic shock (endotoxin)
 It is the target of attack of some enzymes and antibiotics
 It has adhesion function
 Gram negative bacteria
 Very thin peptidoglycan
 No teichoic acids
 Alcohol readily removes the crystal violet.
 Alcohol also dissolves the lipopolysaccharide of the cell wall.
 Gram variable cells
 Some cells retain crystal violet; some decolorize and take up the
safranin
 4 factors-
 Genetics- variable amount of teichoic acid.
 Age of culture- older cultures have variable amount of teichoic acid
 Growth medium- necessary nutrients not available
 Technique-
 smear not thin or evenly made.
 Staining procedure not done correctly- decolorizer left on too long.
 Gram
positive

Gram
negative
 Gram staining is useful because it classifies
whether bacteria is gram-positive or gram-
negative.
 Knowing of gram reaction of bacteria it is
important because it reveals the differences
of bacteria.
 Gram-positive and Gram-negative differ in
how susceptible they are to the antibiotics.
 They produce different toxic materials.
 They react differently to disinfectants.
 Gram nonreactive cells
 Have peptidoglycan but have very waxy- thick lipids –
waterproof, dyes cannot enter either.
 Examples- Mycobacterium tuberculosis and leprosy.
 Alternative staining- acid fast stain
 The Ziehl-Neelsen stain (ZN stain), also called the
hot method of AFB staining, is a type of
differential bacteriological stain used to identify
acid-fast organisms, mainly Mycobacteria.
 Acid fast organisms are those which are capable
of retaining the primary stain when treated with an
acid (fast=holding capacity).
 Members of the Actinomycetes, genus Nocardia
(N. brasiliensis and N. asteroides are opportunistic
pathogens) are partially acid-fast.
 Acid-fast
bacteria have a waxy substance
called mycolic acid in their cell walls which
makes them impermeable to many staining
procedures, including the Gram stain.
 These bacteria are termed "acid-fast"
because they are able to resist
decolorization with acid alcohol.
 Carbol fuchsin is the primary stain in this
procedure, and it contains phenol to help
solubilize the cell wall. Heat is also applied
during the primary stain to increase stain
penetration. All cell types will take up the
primary stain.
 The cells are then decolorized with acid-
alcohol, which decolorizes all cells except
the acid-fast ones.
 Methylene blue is then applied
to counterstain any cells which have been
decolorized.
 At the end of the staining process, acid-fast
cells will be reddish-pink, and non-acid fast
cells will be blue.
 (Note: Acid-fast stains are performed on
smears that have been heat-fixed.)
 L-forms ( Lister Institute where
discovered)
 Bacteria loses cell wall during the life cycle
 Result of a mutation in cell wall forming genes
 Induced by treating with lysozyme or penicillin which
disrupts the cell wall
 Protoplast-
 G + bacterium with no c. wall, only a c. membrane
 Fragile, easily lysed
 Spheroplast-
 G – bacterium loses peptidoglycan, but has outer
membrane
 Less fragile but weakened.
 Located just under cell wall
 Very thin
 Lipid bilayer, similar to the plasma membrane
of other cells. Transport of ions, nutrients and
waste across the membrane
 Typical
 30-40% phospholipids
 60-70% proteins
 Exceptions-
 Mycoplasma- sterols
 Archaea- unique branched hydrocarbons
 The cytoplasmic membrane has a classical
membrane structure: a phospholipid double
layer in which proteins and glycoproteins are
inserted.
 Missing sterols (except myoplasms).
 In gram-positive bacteria, CPM forms
mesosomes (septal, lateral), containing
enzymes, cytochromes and proteins of the
electron transport system.
 PBP de la “penicillin-binding proteins”(they are
inactivated by penicillin and beta-lactam
antibiotics).
 Enzymes that participate in cell biosynthesis to
ensure cell growth
 Permease(components of the transmembrane
transport system)
 Protein that have receptors.
 Respiratory chain proteins (cytochromes)
 Carries out functions normally carried out by eukaryote
organelles.
 Site for energy functions
 Nutrient processing
 Synthesis
 Transport of nutrients and waste
 Selectively permeable
 Most enzymes of respiration and ATP synthesis
 Enzyme synthesis of structural macromolecules
 Cell envelope and appendages
 Secretion of toxins and enzymes into environment.
1.Plasmolysis phenomena (in hypertonic medium - cytoplasm
retraction) or plasmoptysis (in hypotonic medium - lysis of the
cell, separation of MCP from the cell wall)
2. Electronic microscopy
Extension of cell membrane
 Folding into cytoplasm – internal pouch
 Increases surface area.
 Gram-positivebacteria-prominent
 Gram negative bacteria- smaller, harder to see.
 Functions-
 Cell wall synthesis
 Guides duplicated
chromosomes into
the daughter cells
in cell division.
 Encased by cell membrane
 Dense, gelatinous
 Prominent site for biochemical and synthetic
activities
 70-80% water- solvent
 Mixture of nutrients- sugar, amino acids, salts
 Building blacks for cell synthesis and energy
 It lack cellular organelles
Functions: the seat of all metabolic processes
 Highlighting: by any staining method
 Singularcircular strand of DNA
 Aggregated in a dense area- nucleiod
 Long molecule of DNA tightly coiled around
protein molecules.
 A prokaryote is a simple, single-celled organism
that lacks a nucleus and membrane-bound
organelles. The majority of prokaryotic is found in
a central region of the cell called the nucleoid, and
it typically consists of a single large loop called a
circular chromosome.
 The nucleoid can be clearly visualized on
an electron micrograph at
high magnification, where, although its
appearance may differ, it is clearly visible
against the cytosol.
 Sometimes even strands of what is thought to
be DNA are visible. By staining with
the Feulgen stain, which specifically
stains DNA, the nucleoid can also be seen
under a light microscope.
 Nonessential pieces of DNA
 Often confer protection- resistance to drugs

 Tiny, circular
 Free or integrated
 Duplicate and are passed on to offspring
 Used in genetic engineering
 Fertility-F-plasmids. They are capable of
conjugation (transfer of genetic material between
bacteria which are touching).

 Resistance-(R)plasmids, which contain genes that

can build a resistance against antibiotics or poisons and
help bacteria produce pili .

 Col-plasmids, which contain genes that determine the

production of bacteriocins, proteins that can kill other
bacteria.

 Degradative plasmids, which enable the digestion of

unusual substances, e.g., toluene or salicylic acid.

 Virulence plasmids, which turn the bacterium into a

pathogen (one that causes disease).
 Ent plasmids - production of enterotoxin
 Hly plasmids - production of hemolysin
 Degradative plasmids, which enable the digestion
of unusual substances, e.g., toluene or salicylic acid.

 Virulence plasmids, which turn the bacterium into a

pathogen (one that causes disease).
 Prokaryotes use a
relatively simple form
of cell division -
binary fission.
 The diagram at 1.shows a
bacterial cell.
 The cell wall and
membrane are in red,
 the bacterial chromosome
in blue,
 the cytoplasm in light
green,
 the yellow dot represents
a point of attachment of
the chromosome to the
cell membrane.
 Siteof protein
synthesis
 Thousands
 Occurs in chains
–polysomes
 70S
 2 smaller
subunits
 30S and 50S
 “Happiness and bacteria have one
thing in common; they multiply by
dividing!”

 “Ifyou don't like bacteria, you're on

the wrong planet.”