Ryan Danvers

How does the structure of a Gram negative bacterium differ from that of a Gram positive
bacterium? What consequences does this have for human disease?

The fundamental difference between Gram positive and Gram negative bacteria lies within the
composition of the cell wall - and thus the subsequent result produced in Gram staining: a
staining technique using a crystal violet dye that is retained in the presence of peptidoglycan. As
demonstrated by Hans Christian Gram, Gram-positive bacteria constitute mainly peptidoglycan
in their cell wall, staining blue when viewed under the microscope as the dye is retained by this
relatively thick peptidoglycan layer. Whereas, peptidoglycan only constitutes a thin portion of
cell walls in Gram-negative bacteria, whereby the outer membrane of the cell wall is made of a
lipopolysaccharide complex not found in Gram positive bacterium. Thus, these bacteria stain
red/pink with Gram staining as this thinner layer of peptidoglycan insufficiently retains the blue
dye used in Gram staining.

Gram-positive bacteria, such as species in the genera Staphylococcus and Clostridium, contain
several layers of peptidoglycan: a macromolecule consisting of many crosslinked
glycosaminoglycans such as N-acetylglucosamine and N-acetylmuramic acid via short-peptides.
This structure offers protection within Gram-positive bacteria and establishes their shape.
Notably, only partial cross linking is present in Gram-negative bacteria due to there only being a
thin layer of peptidoglycan.

Furthermore, Gram-positive bacteria contain teichoic acids (WTAs) throughout the cell wall.
Significantly, wall-teichoic acids are responsible for sensitivity (and thus resistance) to
Ryan Danvers

antibiotics, such as the B-lactam resistance found in MRSA. As wall teichoic acids are highly
negatively charged and hydrophobic, they modulate binding to extracellular molecules - which
also influences how easily bacteria can enter cells. Experimentally blocking WTA synthesis in B.
subtilis (via KO gene for synthetic enzyme) increases sensitivity to B-lactam antibiotics.
As highly antigenic structures, proteins involved in the WTA biosynthesis pathway are potential
sites for antibiotic development. For instance, targocil is an inhibitor of TarG - an essential
enzyme needed in late stages of synthesis. Experimentally using targocil in Gram-positive
bacteria removes organisms’ capacity to withstand high temperatures nor osmotic stress
exhibitted from hypertonic salt solution also suggesting that WTAs play a role in protecting
Gram-positive bacteria from adverse environments. Furthermore, teichoic acids contribute to
pathogenesis via facilitating adherence to host tissue - such as that found when Staphylococcus
aureus attaches to mucosal cells: emphasising their essentiality in normal Gram-positive
function.

Gram-negative bacteria are structurally different to Gram-positive bacteria: containing only a thin
peptidoglycan layer and a periplasmic space between the inner plasma membrane and outer
membrane containing the lipopolysaccharide (LPS) complex. This complex is an endotoxin
constituting 3 layers: O-antigen (a target for host antibodies and antibiotics), the outer core and
inner core (containing a Lipid A molecule). Notably, the outer membrane plays an essential role
in protecting Gram-negative bacteria against antibiotics (i.e. penicillin), lysosomes and
detergents (which would otherwise damage the inner peptidoglycan), with the LPS especially
playing an essential role in maintaining the selective barrier function. Zhang (2013)
demonstrated that experimentally altering/removing the LPS complex (i.e. via deletion of ZlgC
gene) caused death in many Gram-negative bacteria, demonstrating its central role in defence
and survival.

In the context of human disease, Lipid A within the LPS complex is responsible for the toxicity of
Gram-negative bacteria. Lipid A is an extremely hydrophobic molecule, allowing the LPS to be
securely anchored to the outer membrane. Lipid A activates macrophages and complement
upon entry into human hosts; though, lysis of Lipid A by immune cells induces septic shock.
This manifests as dangerously low hypotension, as well as a fever and a high respiratory rate.
The endotoxic nature of Gram-negative bacteria directly contrasts with the (conventionally)
exotoxic nature of Gram-positive bacteria, such as Staphylococcus aureus. In this instance,
heat labile proteins synthesised during bacterial growth and metabolism are secreted
extracellularly following lysis of the prokaryote itself: rather than the outer membrane constituent
of its cell wall. Clinically, exotoxins are the basis for tetanus and diptheria.
Additionally, O-antigen can also contribute to virulence - evidenced by E.coli O157:H7 which
causes hemolytic uremic syndrome, manifesting as kidney failure due to damage of renal blood
vessels.

The outer membrane is a significant medical challenge, not only as a permeability barrier to
antibiotics, but also due to the fact that the periplasmic space hosts multiple enzymes that
Ryan Danvers

degrade antibiotics. Thus, B-lactam antibiotics typically used on Gram-positive bacteria, such
as vancomycin, are ineffective as they are unable to penetrate and thus reach peptidoglycan
within the cell wall. .

Pili are virtually exclusive to Gram-negative bacteria; though, are present on C. renale (a
Gram-positive species). Pili play a vital role in bacterial transformation in which horizontal gene
transfer occurs: presenting great medical challenge in the evolution of antibiotic resistant strains
of Gram-negative bacteria, such as MRSA and C. difficile.

In conclusion, it is apparent that structural differences exist between Gram-positive and

Gram-negative species have established differences in the way in which each broad category
induces pathogenesis. As our understanding of each bacterium’s structure develops, it opens
opportunity for further development of therapeutics, in the hope to overcome barriers in
penetration and metabolism within the prokaryotic cell-wall structure that are currently enabling
antibiotic-resistance.