Differences between Gram-negative Bacteria and Gram-positive Bacteria

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Gram-positive vs. Gram-negative bacteria

Gram staining is a laboratory method used to differentiate bacterial species into two large groups based on the physical and chemical properties of their cell walls.[1][2] The technique is named after the Danish bacteriologist Hans Christian Gram, who developed the procedure in 1884.[3] The staining process separates bacteria into Gram-positive and Gram-negative groups by identifying differences in the thickness of the peptidoglycan layer in their cell walls.[3][2] Gram-positive bacteria retain the initial crystal violet stain and appear purple, while Gram-negative bacteria lose the initial stain and take up the safranin counterstain, appearing pink or red.[4][5][1]

Comparison Table

Feature Gram-positive Bacteria Gram-negative Bacteria
Gram stain result Retains crystal violet dye; appears purple/blue.[4][5][1] Loses crystal violet; takes up safranin counterstain and appears pink/red.[4][5][1]
Peptidoglycan layer Thick (many layers).[5] Thin (single layer).[5]
Outer membrane Absent. Present, containing lipopolysaccharides (LPS) and porins.[4][5]
Lipopolysaccharide (LPS) Absent. Present in the outer membrane; the lipid A component acts as an endotoxin.
Teichoic acids Present (includes lipoteichoic and wall teichoic acids). Absent.[5]
Periplasmic space Very small or absent. Present, located between the inner and outer membranes.[5]
General antibiotic resistance Generally more susceptible to beta-lactam antibiotics. More intrinsically resistant due to the outer membrane acting as a permeability barrier.[5]
Venn diagram for Differences between Gram-negative Bacteria and Gram-positive Bacteria
Venn diagram comparing Differences between Gram-negative Bacteria and Gram-positive Bacteria


Cell wall structure

The primary structural difference between these two groups of bacteria is the composition of their cell envelope.

A Gram-positive cell wall is characterized by a thick peptidoglycan layer, which can be up to 30 layers thick and constitutes up to 60% of the cell wall mass. This thick mesh is what retains the crystal violet-iodine complex during the decolorization step of the Gram stain.[3][2] Embedded within the peptidoglycan layer are polymers called teichoic acids, which are unique to Gram-positive bacteria. These include wall teichoic acids, which are covalently linked to the peptidoglycan, and lipoteichoic acids, which are anchored to the cytoplasmic membrane.

In contrast, the Gram-negative cell wall is more complex but has a much thinner peptidoglycan layer, often only a single layer thick.[4][5] This thin layer is located within the periplasmic space, an area between the inner cytoplasmic membrane and a distinct outer membrane.[5] Because of this thin peptidoglycan layer, the crystal violet-iodine complex is easily washed out by the decolorizer.[3] The outer membrane is a defining feature, composed of a lipid bilayer where the outer leaflet contains lipopolysaccharide (LPS). The lipid A portion of LPS is an endotoxin, which can trigger a strong immune response in humans.

Clinical significance

The differences in cell wall structure have significant implications for bacterial pathogenicity and treatment. The outer membrane of Gram-negative bacteria provides an additional protective barrier, making them inherently more resistant to many antibiotics, detergents, and lysozyme.[5] This membrane prevents certain drugs from reaching their target within the bacterial cell.

Gram-positive bacteria lack this outer membrane and are typically more susceptible to antibiotics that target the peptidoglycan cell wall, such as penicillin and other β-lactams. However, many Gram-positive pathogens have acquired resistance through other mechanisms, such as altering the antibiotic's target site. The LPS in Gram-negative bacteria is a potent endotoxin that can be released when the bacterial cell is destroyed, leading to symptoms like fever, inflammation, and, in severe cases, septic shock.


References

  1. 1.0 1.1 1.2 1.3 "clevelandclinic.org". Retrieved December 02, 2025.
  2. 2.0 2.1 2.2 "carleton.edu". Retrieved December 02, 2025.
  3. 3.0 3.1 3.2 3.3 "ksu.edu.sa". Retrieved December 02, 2025.
  4. 4.0 4.1 4.2 4.3 4.4 "medicalnewstoday.com". Retrieved December 02, 2025.
  5. 5.00 5.01 5.02 5.03 5.04 5.05 5.06 5.07 5.08 5.09 5.10 5.11 "byjus.com". Retrieved December 02, 2025.