BACTERIAL ENDOTOXINS

ENDOTOXINS

• Endotoxins are part of the outer membrane of the cell wall

of Gram-negative bacteria.

• The term "endotoxin" is used to refer to any cell-associated

bacterial toxin, but in bacteriology it is properly reserved to
refer to the lipopolysaccharide complex associated with the
outer membrane of Gram-negative bacteria.

• Examples:Salmonella, Shigella, Pseudomonas, Neisseria etc.
CHEMICAL STRUCTURE OF ENDOTOXINS

• Lipopolysaccharides are complex amphiphilic

molecules with a molecular weight of about 10kDa.

• LPS can be extracted from whole cells by treatment

with 45% phenol at 90oC. Mild hydrolysis of LPS
yields Lipid A plus polysaccharide.

• LPS consists of three components or regions: Lipid A,

an R polysaccharide and an O polysaccharide.
 Region I : Lipid A

• Lipid A is the lipid component of LPS.

It contains the hydrophobic, membrane-anchoring region of
LPS.

• It consists of a phosphorylated N-acetylglucosamine (NAG)

dimer with 6 or 7 fatty acids (FA) attached. Usually 6 FA are
found. All FA in Lipid A are saturated.

• Some FA are attached directly to the NAG dimer and others

are esterified to the 3-hydroxy fatty acids that are
characteristically present.

• The structure of Lipid A is highly conserved among Gram-

negative bacteria.
 Region II : Core (R) antigen or R polysaccharide

• It is attached to the 6 position of one NAG. The R antigen

consists of a short chain of sugars. For example: KDO - Hep
- Hep - Glu - Gal - Glu - GluNAc -Two unusual sugars,
heptose and 2-keto-3-deoxyoctonoic acid (KDO), are
usually present, in the core polysaccharide. KDO is unique
and invariably present in LPS and so it has been used as an
indicator in assays for LPS.

• With minor variations, the core polysaccharide is common

to all members of a bacterial genus (e.g. Salmonella), but it
is structurally distinct in other genera of Gram-negative
bacteria. 
 Region III : Somatic (O) antigen or O polysaccharide 

• It is attached to the core polysaccharide.

• It consists of repeating oligosaccharide subunits made up

of 3 - 5 sugars. The individual chains vary in length ranging
up to 40 repeat units.

• The O polysaccharide is much longer than the core

polysaccharide, and it maintains the hydrophilic domain of
the LPS molecule. A major antigenic determinant
(antibody-combining site) of the Gram-negative cell wall
resides in the O polysaccharide.
 LPS & VIRULENCE

• Both Lipid A and the polysaccharide side chains act as

determinants of virulence in Gram-negative bacteria.

• LPS elicits a variety of inflammatory responses in an animal

and it activates the complement system by the properdin
pathway.

• Toxicity is associated with the lipid component i.e Lipid A,

whereas immunogenicity is associated with
the polysaccharide component.
 The O polysaccharide and Virulence

• Virulence, and the property of smoothness,  is associated

with an intact O polysaccharide.
• Small changes in the sugar sequences in the side chains of
LPS result in major changes in virulence.
• The O polysaccharide helps in virulence in the following
ways:

1. Allow organisms to adhere specifically to certain tissues,

especially epithelial tissues.

2. Smooth antigens probably allow resistance to

phagocytes, since rough mutants are more readily engulfed
and destroyed by phagocytes.
 Lipid A and Virulence

• The physiological activities of LPS are mediated mainly by

the Lipid A component of the LPS.

• Exerts its toxic effects when released from multiplying cells

in a soluble form, or when the bacteria are lysed.

• Injection of living or killed Gram-negative cells or purified

LPS into experimental animals causes a wide spectrum of
nonspecific pathophysiological reactions, such
as fever, changes in white blood cell counts,disseminated
intravascular coagulation, hypotension, shock and death. 
 ENDOTOXINS vs EXOTOXINS
•  Endotoxins are less potent and less specific in their action,
since they do not act enzymatically.

• Endotoxins are heat stable (boiling for 30 minutes does not

destabilize endotoxin), but certain powerful oxidizing
agents such as superoxide, peroxide and hypochlorite, have
been reported to neutralize them.

•  Endotoxins, although antigenic, cannot be converted to

toxoids.
 References

• Online Textbook of Bacteriology by Dr. Kenneth

Todar.

• www.google.com

• en.wikipedia.org
 Presentation by:
Siddharth Rangnekar
IG-MBT – VIII
Roll no. 29