Antibiotics

Biotechnology II

Antibiotics Disrupt Cell Wall Synthesis,

Protein Synthesis, Nucleic Acid Synthesis
and Metabolism

Univ S. Carolina

Principles and Definitions

Selectivity
Selectivity

vs toxicity

Therapeutic index
Toxic dose/ Effective dose

Categories of antibiotics
Bacteriostatic
Reversibly inhibit growth
Duration of treatment sufficient for host defenses to
eradicate infection

Bactericidal Kill bacteria

Usually antibiotic of choice for infections in sites such as
endocardium or the meninges where host defenses are
ineffective.

Principles and Definitions

Selectivity
Therapeutic index
Categories of antibiotics

Use of bacteriostatic vs bactericidal

antibiotic
Therapeutic index better for bacteriostatic
antibiotic
Resistance to bactericidal antibiotic
Protein toxin mediates disease use
bacteriostatic protein synthesis inhibitor to
immediately block synthesis of toxin.

Principles and Definitions

Antibiotic susceptibility testing (in vitro)

Bacteriostatic Antibiotics
Minimum inhibitory concentration (MIC)
Lowest concentration that results in inhibition of visible
growth (colonies on a plate or turbidity of liquid culture)

Bactericidal Antibiotics
Minimum bactericidal concentration (MBC)
Lowest concentration that kills 99.9% of the original inoculum

Antibiotic Susceptibility Testing-MIC

Disk Diffusion Test
Determination of MIC
Str

Tet

4
2
1
0
Tetracycline (g/ml)
MIC = 2 g/ml

Ery

Chl

Amp

Size of zone of inhibition depends on sensitivity, solubility, rate of diffusion.

Compare results to MIC tables generated using standards.

Principles and Definitions

Combination therapy
Prevent emergence of resistant strains
Temporary treatment until diagnosis is made
Take advantage of antibiotic synergism
Penicillins and aminoglycosides inhibit cell wall synthesis
and allow aminoglycosides to enter the bacterium and
inhibit protein synthesis.
CAUTION: Antibiotic antagonism
Penicillins and bacteriostatic antibiotics. Cell wall synthesis
is not occurring in cells that are not growing.

Antibiotics vs chemotherapeutic agents vs

antimicrobials
Antibiotics-naturally occurring materials
Chemotherapeutic-synthesized in the lab (most
antibiotics are now synthesized and are therefore
actually chemotherapeutic agents.

Mechanisms of Antibiotics
Inhibition of Protein Synthesis
Mostly bacteriostatic
Selectivity due to differences in prokaryotic and
eukaryotic ribosomes
Some toxicity - 70S ribosomes eukaryotic in mitochondria
Inhibitors of INITATION
30S Ribosomal Subunit (Aminoglycosides, Tetracyclines,
Spectinomycin)
50S Ribosomal Subunit (Chloramphenicol, Macrolides)
Inhibitors of ELONGATION
Elongation Factor G (Fusidic acid)

Mechanisms of Antibiotics
Inhibitors of Nucleic Acid Synthesis
Inhibitors of RNA Synthesis: Selectivity due to
differences between prokaryotic and
eukaryotic RNA polymerase: (Rifampcin)
Inhibitors of DNA Synthesis: Selectivity due to
differences between prokaryotic and
eukaryotic enzymes: (Quinolones)
Inhibitors of Folic Acid Synthesis :bacteria make
it, we get it from diet (Sulfonamides)

Antimicrobial Drug Resistance

Principles and Definitions

Clinical resistance vs actual resistance

Resistance can arise by new mutation or by gene
transfer (e.g. acquisition of a plasmid)
Resistance provides a selective advantage.
Resistance can result from single or multiple steps
Cross resistance vs multiple resistance
Cross resistance -- Single mechanism-- closely related
antibiotics are rendered ineffective
Multiple resistance -- Multiple mechanisms -- unrelated
antibiotics. Acquire multiple plasmids. Big clinical
problem.

Antimicrobial Drug Resistance

Mechanisms

Altered permeability
Altered influx

Mutation in a transporter necessary to import antibiotic can lead

to resistance.

Altered efflux

Acquire transporter gene that will pump the antibiotic out

(Tetracycline)

Inactivation of the antibiotic

b-lactamase
Chloramphenicol Acetyl Transferase

Mutation in the target site.

Penicillin binding proteins (penicillins)

RNA polymerase (rifampin)
30S ribosome (streptomycin)

Replacement of a sensitive enzyme with a resistant

enzyme

Plasmid mediated acquisition of a resistant enzyme (sulfonamides,

trimethoprim)