REG. NO: SC/ NUS/ 10/ 0039.

ANNAH, ITSUKO KAI.

26th August, 2010.
NUR 229; MICROBIOLOGY.

Assignment 1: Describe the detail structure of a Bacterial cell wall and indicate how
different its chemical composition is from Plant and Fungi cell wall.

A cell wall is a tough, usually flexible but sometimes fairly rigid layer that surrounds some
types of cells. It is located outside the cell membrane and provides structural support and
protection and also act as a filtering mechanism.
In bacterial cell wall is rigid to gives it a definite shape, appears porous which forms an
electron-dense layer outside the plasma membrane. It is made up of polysaccharides, protein and
frequently also lipids. It is 10 to25u thickness and weighs 20% of total weight of the cell. Its
chemical nature varies from species to species, the principal component of bacterial cell wall is
peptidoglycan [it is a polymer or interlocking chains of identical monomers and composes of two
derivatives of glucose; N-acetylgucosamine and N-acetylmuramic acid, this glucose strands are
connected by interpeptide bridges]. Glutamic acid, lysine, diaminopimellic, teichoic acid,
homopolymers of peptides, lipoproteins and lipopoly-saccharides are also present in the bacterial
cell wall. Many bacterial cell wall are surrounded by an out gelatinous capsule which is tightly
bound to the cell wall, When the capsular coat is free from the cell wall it is called slime. The
capsule makes the bacterial pathogenic, the slime layer compose of polymers of carbohydrate
and display various morphological patterns.
The structure of bacterial cell wall divides bacterial into two broad classes of that can be
distinguish by a staining procedure called Gram stain; the Gram-positive bacterial cell wall ,
contains 30-40 peptidoglycan layers and can be lysed by the enzyme lysozyme which have the
property of dissolving the cell wall by breaking the 1-4glycoside bond and renders the organism
naked, while in the Gram- negative bacterial cell wall contains 1-3 layers of peptidoglycans
surrounded by second rigid membrane of lipopolysaccharide [which are hydrophilic in nature
giving the wall surface a soft and smooth to act as deterrent for engulfment by phagocytic cells]
and lipoproteins. The cell wall also contains antigens to generate immunity in vertebrates.
In contract the major carbohydrate making the primary plant cell wall are cellulose, pectin,
hemicelluloses, while chitin forms that of fungi. The cellulose micro fibrils are linked via
hemicellulosic tethers to form a network which via embedded in the pectin matrix in the plant
cell wall. The outer part of the primary cell wall of plant epidermis is compose of cutin and wax
forming a permeability barrier called plant cuticle, the secondary cell wall contain compound
that modify their mechanical properties and permeability, the polymer that make up wood
include cellulose, xylan and a complex phenotic polymer called lignin.

References:
S, C. Rostogi, Cell and Molecular Biology, second edition. Pp 5-8, 120
Purves, Sadava, Orians, Heller,
Website: Bauman, R. (2005) Microbiology.
Park Talaro, K. (2008) Foundation of Microbiology
REG. NO: SC/ NUS/ 10/ 0039
ANNAH, ITSUKO KAI
26th August, 2010.
NUR 229: MICROBIOLOGY

Assignment 2: Describe the Gram-stain technique and indicate the differences between Gram-
negative and G ram-positive bacterial with special reference to the composition of their cell wall.

Christian Gram, a Danish bacteriologist in the 1800”s developed the bacterial staining
technique. This involves the application of series of dyes, specific stain reaction of the bacterium
results from the structure of its cell wall. Most bacterial have one of these two types of cell wall,
the differential Gram stain uses two dyes to distinguish between the bacterial cell wall types.
Before staining a bacterial, smear must be prepared and heat fixed to a microscope slide. A
smear is a sample of bacterial suspended in a small amount of water on a slide, the sample is then
dried using heat. The heat kills the bacterial and attaches the sample to the slide so that it does
not easily wash away.
Procedure:
-The slide is flood with crystal violet [primary stain]
-After one minute the slide is rinse with water.
-Flood the slide with iodine [a mordant that binds the crystal violet to the Gram cell postive wall]
-After one minute rinse the slide with water.
-Flood the slide with acetone alcohol [alcohol is a decolorizer that will remove the stain from the
Gram negative cell.
-After 10 to 15 seconds, rinse the slide with water [do not leave the decolorizer on too long or it
may remove stain from the Gram positive cell wall.
-Flood slide with safrinin [the counterstain]
-After one minute, rinse slide with water.
-Gently blot slide dry. It is now ready to be viewed under oil immersion [1000x TM] with a
bright-field compound microscope.
After this staining procedure, the Gram positive cell will appear purple, due to the complex
called peptidogylcan layer been thicker in the Gram positive bacterial cell wall having retained
the primary stain. The Gram negative will appear pink, due to the location of cell wall
peptidoglycan and lipopolysaccharide and have retained the counterstain after the primary stain
was removed by the decolorizer.
The vast majority of Gram negative is pathogens, bacterial that can cause diseases. This
pathogenicity is typically associated with lipopolysaccharide endotoxins in Gram negative wall;
it can also produce toxicity in a host organism, and in human may trigger a nonspecific immune
response marked by the production of cytokines which result in inflammation. The lipids protect
the Gram negative bacterial cell wall membrane to resist antibiotics to which Gram positive
bacterial cell wall are susceptible.

Reference:
Poger, Y. stanier, Michael, Daidoroff, Edward, A. Adelbo
Website: Bauman, R. [2005] Microbiology.
Park Talaro, K. [2008] Foundation of microbiology.