Structure and Synthesis of

Peptidoglycan
Lecture # 1
Microbial anatomy and physiology
Dr. Qasim Mughal
05-06-2023
Structure of Peptidoglycan
• Peptidoglycan, or murein, is an enormous meshl-ike polymer composed
of many identical subunits.
• The polymer contains two sugar derivatives,
– N-acetylglucosamine and
– N-acetylmuramic acid (the lactyl ether of N-acetylglucosamine), and
– several different amino acids.
• Three of these amino acids are not found in proteins:
– D-glutamic acid,
– D-alanine, and
– mesodiaminopimelic acid.
• The presence of D-amino acids protects against degradation by most
peptidases, which recognize only the L-isomers of amino acid residues.
Structure
• The backbone of this polymer is composed of alternating N-
acetylglucosamine and N-acetylmuramic acid residues.
• A peptide chain of four alternating D- and L-amino acids is
connected to the carboxyl group of N-acetylmuramic acid.
• Many bacteria replace meso-diaminopimelic acid with another
diaminoacid, usually L-lysine
• Figure: Peptidoglycan Subunit
Composition.
• The peptidoglycan subunit of E. coli,
most other gram-negative bacteria,
and many gram-positive bacteria.
– NAG is N-acetylglucosamine.
– NAM is N-acetylmuramic acid (NAG
with lactic acid attached by an ether
linkage).
– The tetrapeptide side chain is composed
of alternating D- and L-amino acids since
meso-diaminopimelic acid is connected
through its L-carbon.
– NAM and the tetrapeptide chain attached
to it are shown in different shades of
color for clarity
• Figure: Peptidoglycan Cross-Links.

– (a) E. coli peptidoglycan with direct

cross-linking, typical of many gram-
negative bacteria.
– (b) Staphylococcus aureus peptidoglycan.
S. aureus is a gram-positive bacterium.

– NAM is N-acetylmuramic acid.

– NAG is N-acetylglucosamine.
– Gly is glycine.
– Although the polysaccharide chains are
drawn opposite each other for the sake
of clarity, two chains lying side-by-side
may be linked together
Figure 3.21 Peptidoglycan Structure. A schematic diagram of one
model of peptidoglycan. Shown are the polysaccharide chains,
tetrapeptide side chains, and peptide interbridges.
• In order to make a strong, mesh-like polymer, chains of linked
peptidoglycan subunits must be joined by cross-links between the
peptides.
• Often the carboxyl group of the terminal D-alanine is connected
directly to the amino group of diaminopimelic acid, but a peptide
interbridge may be used instead.
• Most gram-negative cell wall peptidoglycan lacks the peptide
interbridge. This cross-linking results in an enormous peptidoglycan
sac that is actually one dense, interconnected network
• These sacs have been isolated from gram-positive bacteria and are
strong enough to retain their shape and integrity, yet they are
relatively porous, elastic, and somewhat stretchable
Synthesis of Peptidoglycan
• Nucleoside diphosphate sugars also participate in the synthesis of
peptidoglycan.
• Recall that peptidoglycan is a large, complex molecule consisting of
long polysaccharide chains made of alternating N-acetylmuramic
acid (NAM) and N-acetylglucosamine (NAG) residues.
• Pentapeptide chains are attached to the NAM groups. The
polysaccharide chains are connected through their pentapeptides or
by interbridges
• Not surprisingly, such an intricate structure requires an equally
intricate biosynthetic process, especially because some reactions
occur in the cytoplasm, others in the membrane, and others in the
periplasmic space.
• Peptidoglycan synthesis involves two carriers.
• The first, uridine diphosphate (UDP) functions in the cytoplasmic
reactions.
• In the first step of peptidoglycan synthesis, UDP derivatives of N-
acetylmuramic acid and N-acetylglucosamine are formed.
• Amino acids are then added sequentially to UDP-NAM to form the
pentapeptide chain.
• NAM-pentapeptide is then transferred to the second carrier,
bactoprenol phosphate, which is located at the cytoplasmic side of the
plasma membrane.
• The resulting intermediate is often called Lipid I. Bactoprenol is a 55-
carbon alcohol and is linked to NAM by a pyrophosphate group.
• Next, UDP transfers NAG to the bactoprenol-NAM-pentapeptide
complex (Lipid I), to generate Lipid II. This creates the peptidoglycan
repeat unit.
• The repeat unit is transferred across the membrane by bactoprenol.
• If the peptidoglycan unit requires an interbridge, it is added while the repeat
unit is within the membrane.
• Bactoprenol stays within the membrane and does not enter the periplasmic
space.
• After releasing the peptidoglycan repeat unit into the periplasmic space,
bactoprenol-pyrophosphate is dephosphorylated and returns to the
cytoplasmic side of the plasma membrane, where it can function in the next
round of synthesis.
• Meanwhile, the peptidoglycan repeat unit is added to the growing end of a
peptidoglycan chain.
• The final step in peptidoglycan synthesis is transpeptidation, which creates the
peptide cross-links between the peptidoglycan chains. The enzyme that
catalyzes the reaction removes the terminal D-alanine as the cross-link is
formed.
• To grow and divide efficiently, a bacterial cell must add new
peptidoglycan to its cell wall in a precise and well-regulated way
while maintaining wall shape and integrity in the presence of high
osmotic pressure.
• Because the cell wall peptidoglycan is essentially a single,
enormous network, the growing bacterium must be able to
degrade it just enough to provide acceptor ends for the
incorporation of new peptidoglycan units.
• It must also reorganize peptidoglycan structure when necessary.
This limited peptidoglycan digestion is accomplished by enzymes
known as autolysins, some of which attack the polysaccharide
chains, while others hydrolyze the peptide cross-links. Autolysin
inhibitors are produced to keep the activity of these enzymes
under tight control.
• Although the location and distribution of cell wall synthetic activity varies
with species, there seem to be two general patterns.
• Many gram-positive cocci (e.g., Enterococcus faecalis and Streptococcus
pyogenes) have only one to a few zones of growth.
• The principal growth zone is usually at the site of septum formation, and new
cell halves are synthesized back to-back.
• The second pattern of synthesis occurs in the rod-shaped bacteria Escherichia
coli, Salmonella, and Bacillus.
• Active peptidoglycan synthesis occurs at the site of septum formation, but
growth sites also are scattered along the cylindrical portion of the rod.
• Thus growth is distributed more diffusely in rod-shaped bacteria than in the
streptococci.
• Synthesis must lengthen rod-shaped cells as well as divide them. Presumably
this accounts for the differences in wall growth pattern
• Because of the importance of peptidoglycan to cell
wall structure and function, its synthesis is a
particularly effective target for antimicrobial
agents.
• Inhibition of any stage of synthesis weakens the
cell wall and can lead to osmotic lysis.
• Many commonly used antibiotics interfere with
peptidoglycan synthesis.
• For example, penicillin inhibits the
transpeptidation reaction, and bacitracin blocks
the dephosphorylation of bactoprenol
pyrophosphate