LIPID-CELL WALL

IDENTIFICATION
Carl G. Buscato and Riza MaeEayte
Bio 135
Republic of the Philippines
College of Natural Science and
Mathematics
MINDANAO STATE UNIVERSITY
General Santos City
Classifications Schemes to
Consider
Gram Reaction
Gram Positive Gram Negative
Acid
Fast
Non-acid Fast
Gram Positive Acid Fast
Gram Positive Non-Acid Fast

Mycolic acid (60%)

Have lipids linked to peptidoglycan

Includes the genera of:
Mycobacteria (C
60-90
)
Nocardia (C
46-58
)
Rhodococcus (C
28-48
)
Corynebacterium (C
24-36
)
Gram Positive Acid Fast
Thick, multi-layered peptidoglycan

Absence of lipid outer membrane

Low lipid, lipoprotein and lipopolysaccahride
content

Produces exotoxins
Gram Positive Non-Acid Fast
Gram Negative
Thin, single-layered peptidoglycan layer

Presence of lipid outer membrane

High lipid, lipoprotein and lipopolysaccharide
content

Produces endotoxin (Lipid A) and exotoxins

Since, gram negative have high lipid content,
the lipid-cell wall taxonomic scheme best fits
for them

Gram Negative
Lipids
Water-soluble (polar) organic molecules that
are important for the structure of the
cytoplasmic membrane and the cell wall.
Lipids are a useful non-genetic criteria to
differentiate Archaea from Bacteria.
Classified into:
Phospholipid
Phosphatyl-glycerol
Di-phosphatyl-glycerol
Phosphatyl-ethanolamine
Phosphatyl-choline
Lipid Classification
Phospholipid
Lipids containing a substituted P-group and 2
fatty acid chains on a glycerol backbone. Archaea
and Bacteria are distinguished by the bond
between the FA tail and the glycerol head; ether-
bond (Archae), esther-bond (Bacteria)

Lipid Classification
Phosphatyl-glycerol
Phospholipid with an attached gylcerol molecule
CHOH(CH2OH)2 to the P-group
Distinguishes microbes according to their
configuration: diacyl (I 60% in E. coli), alklacyl (
20% in Salmonella typhirum) and alkenacyl (30%
in Corynebacterium amycolatum)
Phosphatyl-choline
Phospholipids with an attached choline molecule
OH(CH2)2N(CH3)3 to P-group
Phosphatyl-ethanolamin
Phospholipids with an extra ethanolamine
molecule OH(CH2)2NH3+ to the P-group
Di-phosphatyl-glycerol (cardiolipin)
Two phospholipids bound together by a glycerol
molecule CHOH(CH2OH)2

Lipid Classification
Lipid Classification
Peptidoglycan Classification
Fatty Acid Classification
Branched
anteiso, asymmetrically at different sites
branched
iso, symmetrically branched
methyl, H swapped with CH3 group.
Cyclopropane
FA-chain is interrupted by a ring of C-atoms
(C3H8).
Hydroxy
H swapped w/ OH group of some FA-C-atoms;
e.g.: 2-OH,: 3-OH; in Archaea periodically
branched CH3 side chains.
Saturated
No double bond b/w any C of the FA-chain; all C-
atoms are saturated w/ H-atoms
Poly-Unsaturated FA
Have one or more positions along the FA-polymer
chain where 2 adjacent C are linked by double
bond (2 shared pairs of electrons); consequently,
fewer Hs are bonded to the Cs.

Fatty Acid Classification
Chemotaxonomy
Phenotypic analysis based on classification of
bacterial cell wall constitute (chemical markers
such as lipids, proteins and sugars)
Classification and identification can be done
through differences on distribution,
configurations and absence/presence of
molecules per species/genera
Chemotaxonomic methods include:


Chemotaxonomic Methods: Thin
Layer Chromatography
The classic method of
polar lipid extraction
utilizes a monophasic
mixture of chloroform,
methanol and water
for extraction.
2D thin layer
chromatography can
be used to determine
simple 2-dimensional
patterns of polar lipids
which may be
characteristic of
individual taxa.
Chemotaxonomic Methods: Fatty
Accid Methyl Ester (FAME)
Analysis
Chemotaxonomic Methods: MALDI-
TOF MS
Chemotaxonomic Methods: MALDI-TOF
MS
Chemotaxonomic Methods: MALDI-
TOF MS
MALDI-TOF MS: (Mass
Spectronomy)
MALDI-TOF MS: (MALDI)
MALDI-TOF MS: (TOF)
Thought to ponder: