C L I N I C A L B A C T E R I O L O G Y (L E C T U R E)

• Peptidoglycan is composed of an alternating

disaccharide, acetyl glucosamine and acetyl-
Antimicrobials and their Mechanism of muramic acid.
Action ➢ A short peptide chain is attached to the n-
acetyl muramic acid component that
Antibiotics mediated cross-linking with parallel glycan
• Chemical substances that are produced by molecules. The cross-linking provides
microorganisms that has the capacity to inhibit or kill other strength to the mature cell, and this is
catalyzed by transpeptidase enzymes, also
microorganisms.
called penicillin binding proteins or
• Can also be synthesized chemically.
PBT’s.
• May be classified as broad spectrum (wide range or
• The beta-lactam antibiotics form covalent
different kinds of organisms) or narrow spectrum
complexes with enzymes that generate mature
(effective only against a limited numbers of pathogens)
peptidoglycan molecule (transpeptidase enzymes
• Based on origin they may be classified as either natural,
that mediate peptidoglycan cross-linking).
semi-synthetic, or synthetic drugs.
• The action of beta-lactam antibiotics is to bind
➢ Natural antibiotics- those that are produced by
these enzymes preventing peptidoglycan cross-
microorganism such as bacteria or fungi.
linking.
➢ Semi-synthetic antibiotics- those that are
• The active component of the beta-lactam family of
chemical modified from natural drugs.
➢ Synthetic antibiotics- those that produce purely drugs is the beta-lactam structure/ ring.
chemical means. • Glycopeptides inhibit cell wall synthesis by binding
• Based on the effect in microbial agents may be either to the end of the peptidoglycan, preventing
bacteriostatic or bactericidal. transpeptidation/ cell wall synthesis.
➢ Bacteriostatic agents- inhibit the growth of
bacteria but generally do not kill microorganism.
➢ Bactericidal- those that usually kill or destroy
organism and are used for treating life-
threatening infections.

Terminologies
• Minimum-inhibitory concentration (MIC)– the lowest
concentration/ dilution of a drug that can inhibit bacterial
growth.
• Minimum- bactericidal concentration (MBC)– the
lowest concentration/ dilution of a drug that can still kill
bacteria.
• Therapeutic index– ratio of toxic dose to the therapeutic
dose; the higher the index, the more effective the
chemotherapeutic/ safe agent; used to determine relative
safety of a drug.

Characteristics of Antimicrobial Agents

• The agent must be in active form.
• The agent must be able to achieve concentration at the
site of infection that is higher than the pathogen’s MIC to 2. Inhibitors of Folate Synthesis
be effective. Otherwise, the antimicrobial agent will be • Synthesis of Folate is important in the production of
ineffective as the pathogens will still thrive in the infected purines, a component of the DNA
area. • The folic acid pathway provides essential precursor
• Must have selective toxicity. It is preferable to choose molecules needed for DNA biosynthesis
antibiotics that are toxic against microorganisms but do • Sulfamethoxazole/ SMZ (a sulfonamide) prevents
not cause harm to the patient. the formation of dihydropteroate through
competitive inhibition of para-aminobenzoic acid
with the enzyme dihydropteroate synthase
• Trimethoprim blocks the formation of the
tetrahydrofolate by inhibiting dihydrofolate
reductase
• The effect of competitive binding by these two
antibiotics is the cesation of the conversion of para-
aminobenzoic acid to purines disrupting DNA
synthesis

• Some groups of antibiotics target the synthesis of the

cell wall, a critical component of the bacterial cell that
protects it from the environment and osmotic damage.
• Some inhibits synthesis of proteins by attacking the
ribosomes and its components.
• Others inhibits the synthesis of nucleic acids while other 3. Agents that interfere with DNA replication
• DNA replication- a process in which DNA is
interfere with the production of essential metabolites. duplicated in preparation for cell division
• Topoisomerases I, II, III, and IV are necessary for
Types of Antimicrobial Agents DNA replication
1. Cell wall inhibitors • Quinolones are antibiotics that target these
• Selective antibiotics with high therapeutic index enzymes and prevent the process. It affect DNA
• Interferes with the biosynthesis of peptidoglycan. replication by targeting Topoisomerases II (DNA
gyrase) in gram negative bacteria and
Topoisomerase IV in gram-positive bacteria.
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 4. Agents that interfere with DNA transcription • Antibiotics must be able to penetrate the cell wall to reach
• DNA transcription or the synthesis of RNA using a their targets.
strand of DNA as a template ➢ In gram negative bacteria, the negatively
• Transcription of DNA into RNA is mediated by RNA charged LPS functions as a selectively
polymerase. permeable barrier against negatively charged
• Rifampin targets the RNA polymerase subunit, antibiotics; they repels negative-charged
blocking RNA chain elongation antibiotics
5. Agents that inhibit protein synthesis ➢ Porins are protein channels through which
• Some antibiotics target the 30S ribosomal subunit molecules may enter and leave the cell; they
(aminoglycosides, spectinomycin, tetracyclines, serve as outer membrane channels that permit
glycylcyclines) resulting in the misreading of the the influx of nutrients and efflux of waste
mRNA products.
• Others target the 50S ribosomal subunit Efflux
(macrolides, lincosamides, chloramphenicol,
• Efflux pumps are found in bacteria and function as
oxazolidinones, streptogramins) resulting in the
transporter proteins for the extrusion of toxic substances
inhibition of peptide chain elongation.
and antibiotics from the interior of the cell to the external
• Only aminoglycosides are bactericidal; the rest are environment.
bacteriostatic Enzymatic inactivation
• Aminoglycosides may cause deathness and loss • Bacteria can produce enzymes that destroy antimicrobial
of balance, Chloramphenicol may cause agents before they are able to reach the targets
temporary or permanent depression of the bone
• This is commonly encountered in the resistance against
marrow that leads to aplastic anemia and
beta-lactam antibiotics (all contains the beta-lactam ring
leukopenia, and Tetracycline in increase dosage
structure)
may lead to liver and kidney damage and yellowing
• Beta-lactamases are enzymes that hydrolyzes beta-
of teeth in chidre have side effects.
lactams, preventing the antibiotic’s ability to bind
transpeptidase enzymes
• The effect can be minimized by the use of beta-
lactamase inhibitors (clavulanic acid, sulbactam, and
tazobactam). These inhibitors bind beta-lactamases and
reduces their detrimental effects on the beta-lactam
antibiotics
• Penicillioc acid does not have any antimicrobial activity

Target site modification

• Modification of a target can reduce the binding affinity of
the antibiotic to the target
• Quinolones target DNA gyrase (topoisomerase II) and
topoisomerase IV, inhibiting DNA synthesis. Resistance to
quinolones are caused by mutations encoding amino acid
changes in these DNA topoisomerases, which has
reduced affinity to quinolones.
• Mutations may lead to alteration in transpeptidase
enzymes, resulting to decreased affinity to beta-lactam
antibiotics
Acquisition of new targets
• Microorganisms become resistant by acquiring cellular
targets with reduced affinity for the antibiotic
• There is a gene that encodes for a new type of
transpeptidase enzyme with reduced affinity for beta-
lactam antibiotics.
• Sulfonamides compete with dihydropteroate synthase
enzymes to block the formation of nucleotide precursors.
Some bacteria acquired a new enzyme that is unaffected
by sulfonamides

Bacterial Resistance to Antimicrobials

Impermeability

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