Antibacterial Agents: Basis
of Action
Ian Chopra, University of Leeds, Leeds, UK
David Greenwood, University of Nottingham, Nottingham, UK
Antibacterial agents alleviate infection through a variety of modes of action including the
inhibition of protein, nucleic acid and cell wall synthesis.
History of Antibacterial Agents
Folk remedies have been used since ancient times in an
effort to alleviate infection. Most have taken the form of
salves or poultices applied to infected areas of the skin or
body orifices. Some were bizarre concoctions of substances
ranging from dried and powdered reptiles to bats’ dung;
others consisted of herbal preparations or fungal growths
that may, indeed, have had some antibiotic activity. Honey
was widely used and is still recognized as being useful when
applied to skin ulcers. Before the twentieth century,
however, proven treatments for systemic infection were
restricted to a few plant extracts active against parasitic
worms or protozoa, most notably cinchona bark, which
contains the alkaloids, quinine and quinidine, that have
been spectacularly successful in combating malaria since
the seventeenth century. Among antibacterial agents, the
only compounds offering even minimal benefit were
mercury (for syphilis), chaulmoogra oil (for leprosy) and,
since 1895, hexamine (methenamine) for cystitis. All this
changed in 1935, when Gerhard Domagk (1895–1963)
published his epoch-making paper on Prontosil, the
forerunner of the sulfonamides.
Sulfonamides and other synthetic agents
Prontosil was one of a series of dyes that the German
dyestuffs industry was seeking to exploit as antimicrobial
agents following the pioneering work of Paul Ehrlich
(1854–1915) on the differential affinity of dyes for tissues
and microbes. In Prontosil a sulfonamide group was linked
to a red dye on the grounds that this substituent might
enhance its affinity for bacterial cells, as it was known to do
for fibres. In fact, it turned out that Prontosil was split in
the bloodstream, liberating sulfanilamide, the antimicro-
bial activity of which was unsuspected. The dye itself had
no useful activity, so the discovery came about by an
amazing piece of luck.
Domagk reported an astonishing effect of the dye in
protecting mice from an otherwise lethal infection with
haemolytic streptococci. This was quickly confirmed in the
clinic, notably by Leonard Colebrook (1883–1967), who
established its effectiveness in young women with life-
ENCYCLOPEDIA OF LIFE SCIENCES / & 2001 Nature Publishing Group / www.els.net
Introductory article
Article Contents
. History of Antibacterial Agents
. Bacterial Structure and the Basis of Selective Toxicity
. Inhibitors of Bacterial Cell Wall Synthesis
. Inhibitors of Bacterial Protein Synthesis
. Inhibitors of Nucleic Acid Synthesis
. Antifolate Compounds
. Membrane Disorganizing Agents
. Antimycobacterial Drugs
. Future Directions
threatening ‘childbed fever’. However, the compound
nearly fell into disrepute: one formulation marketed in
the USA, ‘Elixir sulfanilamide’, which contained diethy-
lene glycol, killed 105 people. This tragedy led to the 1938
Federal Food, Drug and Cosmetic Act, which required
proof of the safety of new drugs.
From 1939, numerous derivatives of sulfanilamide were
prepared and other antibacterial chemicals were subse-
quently synthesized. These included the antituberculosis
agents, p-aminosalicylic acid (1946), isoniazid (1952) and
pyrazinamide (1952), as well as the first nitrofurans (1944),
nalidixic acid (1962; the first of the quinolone drugs) and
trimethoprim (1967). However, it was the discovery of the
potent antibacterial activity of natural products – anti-
biotics in the true sense of the word – that set the seal on the
therapeutic revolution in the management of infection.
Penicillins and other b-lactam antibiotics
In the same year in which Domagk published the
description of Prontosil, Howard Florey (1898–1968), an
Australian doctor, was appointed Professor of Pathology
at the University of Oxford. While setting up his research
there, a young German refugee, Ernst Chain (1906–1979),
was recommended to him as a scientific collaborator and
he set Chain the task of examining the properties of
naturally occurring bacteriolytic substances, among which
were two compounds, lysozyme and penicillin, which had
been discovered by Alexander Fleming (1881–1955) at St
Mary’s Hospital in London. Fleming had published his
description of penicillin, the product of a contaminant
mould, in 1929, but had taken only a passing interest in the
possibility that it might be used therapeutically. Chain
partially purified the compound and, in 1940, the Oxford
group established its remarkable activity, initially in a
speculative experiment with just eight mice (including four
untreated controls) infected with Streptococcus pyogenes.
Successful human trials followed and were published in
1941. The development of penicillin was beyond the
1
 Antibacterial Agents: Basis of Action
capacity of Britain during World War II, so Florey and his
colleague Norman Heatley took the idea to the United
States, where rapid progress was made in preparing
penicillin on a commercial basis. In 1959, scientists at
Beecham Research Laboratories discovered the penicillin
nucleus, 6-aminopenicillanic acid, in fermentation pro-
ducts, and this paved the way for the development of
methicillin, cloxacillin, ampicillin and the other semisyn-
thetic penicillins.
The cephalosporins were also developed at Oxford. An
antibiotic-producing mould that had been found by
Guiseppe Brotzu in a sewage outfall in Cagliari, Sardinia,
was sent through a British intermediary to the Oxford
group. There it was discovered that the mould produced a
type of penicillin and a steroid-like antibiotic, neither of
which was developed further. The forerunner of the
cephalosporins, cephalosporin C, was detected as a minor
component by further fractionation of the antibiotic
complex. As with penicillins, semisynthetic derivatives
were developed and eventually gave rise to the large family
of compounds that are available today.
It later became clear that antibacterial compounds
containing a b-lactam ring are widely distributed in nature.
Several, including the carbapenems, monobactams and the
b-lactamase inhibitor, clavulanic acid, were subsequently
developed for therapeutic use.
Other antibiotics
In 1939, Rene Dubos (1901–1982) described tyrothricin,
later shown to be a mixture of two antibiotics, tyrocidine
and gramicidin, as a product of a soil bacterium Bacillus
brevis. These compounds proved too toxic to be used, other
than in topical preparations, but Dubos’ former teacher,
the soil microbiologist Selman Waksman (1888–1973) was
prompted to embark on a systematic search for antibiotic
substances in soil microorganisms. A breakthrough came
in 1944, when Waksman’s student, Albert Schatz, dis-
covered streptomycin, the first antibiotic to exhibit
therapeutic activity in tuberculosis.
The therapeutic revolution signalled by the success of
sulfonamides, penicillin and streptomycin stimulated the
pharmaceutical industry to embark on massive screening
programmes for new antibiotics, which soon bore fruit in
chloramphenicol (1947), chlortetracycline (1948), erythro-
mycin (1952), vancomycin (1956), lincomycin (1962) and
gentamicin (1963), among others.
Subsequent developments have chiefly centred on the
production of semisynthetic derivatives of earlier anti-
bacterial compounds. The aim has been to produce
derivatives with improved properties, particularly to
overcome resistance, which has followed in the wake of
all agents that have been brought into therapeutic use. At
the end of the twentieth century, about 250 antibacterial
compounds are on the world market for the treatment of
2 ENCYCLOPEDIA OF LIFE SCIENCES / & 2001 Nature Publishing Group / www.els.net
systemic infection. Most belong to the main families – b-
lactam agents, aminoglycosides, tetracyclines, macrolides,
sulfonamides and quinolones. The challenge for the next
century is to find new classes of compound to forestall the
ever-present threat of resistance to the older agents.
Bacterial Structure and the Basis of
Selective Toxicity
The guiding principle of all antimicrobial chemotherapy is
‘selective toxicity’: to inhibit or kill the infecting organism,
without harming the host. Most antibacterial agents are
able to exploit differences in the structure or metabolism of
bacterial and mammalian cells in order to achieve their
selective effect, and the unwanted side effects of anti-
bacterial therapy generally arise through idiosyncratic
effects unrelated to the primary mechanism of antibacterial
action.
The main structures of the bacterial cell that provide
targets for the selective action of antibacterial agents are
shown in Figure 1.
Bacterial cell envelope
Most common bacterial pathogens can be divided into two
groups – Gram-positive and Gram-negative – according to
their reaction in the Gram stain, which reflects basic
differences in the structure of bacterial cell envelopes. The
cell envelope of both Gram-positive and Gram-negative
bacteria contains peptidoglycan (syn. mucopeptide; mur-
ein). This consists of molecules of N-acetylglucosamine
alternating with N-acetylmuramic acid units that are
substituted with short peptides. Crosslinking of these
Outer membrane Cell wall peptidoglycan
(Gram-negative organisms)
Cell membrane Chromosome
(DNA synthesis
and transcription)
Cytoplasm Ribosome
(metabolic transformations) (protein synthesis)
Figure 1 Bacterial cell, showing possible targets for selective attack.
 Antibacterial Agents: Basis of Action

peptides (through an interpeptide bridge in Gram-positive

bacteria) maintains the shape of the organism and gives the Capsule
cell wall its strength. Since peptidoglycan is absent from
mammalian cells it provides a useful target for selectively
toxic agents, and many useful compounds, including
penicillins, cephalosporins and other b-lactam agents, as LPS
Porin
well as glycopeptides and fosfomycin, act on this structure.
Gram-positive bacteria (staphylococci, streptococci,
clostridia, etc.) have a thick (c. 30 nm) peptidoglycan layer, Outer membrane
which is extensively crosslinked, and is interspersed with
polysugarphosphates – teichoic and lipoteichoic acids P PL
(Figure 2). In contrast, the peptidoglycan of Gram-negative Periplasm
bacteria (Escherichia coli, salmonellae, pseudomonads, P
LP
meningococci, etc.) is thin (c. 2–3 nm) and loosely cross- Peptidoglycan
linked. There is no teichoic or lipoteichoic acid. External to P PL
the peptidoglycan, Gram-negative bacteria have a mem-
brane-like structure, consisting of lipoprotein and lipopo- Inner membrane
lysaccharide, which is selectively permeable and thus
regulates access to the underlying structures (Figure 3).
This outer membrane contains hydrophilic pores (‘porins’)
through which small molecules, including many antibio-
tics, can enter the cell, depending on their molecular Figure 3 The cell envelope of a Gram-negative bacterium. LP,
lipoprotein; LPS, lipopolysaccharide; P, protein; PL, phospholipid. Flagella
weight, stereochemistry and ionic charge. It is this outer are not illustrated. Reproduced with permission from Hancock I and Poxton
membrane that often determines the spectrum of activity of I (1988) Bacterial Cell Surface Techniques. London: John Wiley & Sons.
antibacterial compounds.

Mycobacteria mer. This impermeable outer coat renders the mycobacter-

The mycobacterial cell wall differs from more conventional ia resistant to many antibacterial compounds, and
bacteria in that the peptidoglycan is covalently linked to a specialized antimycobacterial agents, some of which
waxy mycolic acid layer via an arabinogalactan copoly- inhibit mycolic acid formation, have been developed for
use in therapy.

Capsule Cytoplasmic membrane

Like all cells, bacteria have an inner membrane within
which the cytoplasm is contained, but this is insufficiently
Protein layer different from the mammalian cell membrane to be a useful
target for selective attack, and, among systemically-useful
agents, only the polymyxins act at this site.
Peptidoglycan

Bacterial ribosomes
2° wall polymer Many antibacterial agents, including most naturally-
occurring antibiotics derived from soil microorganisms,
PL P
Lipocarbohydrate act by inhibiting various stages in bacterial protein
Cytoplasmic synthesis. The general mechanism of protein synthesis on
membrane
the ribosomal template is similar in all cells, but bacterial
ribosomes are sufficiently different to allow considerable
scope for selective action.
Bacterial ribosomes, like their mammalian counter-
Figure 2 The cell envelope of a Gram-positive bacterium. Note that not all parts, consist of two subunits, each consisting of a complex
the components shown occur in every strain of Gram-positive bacterium. P, of ribosomal ribonucleic acid (RNA) and a number of
protein; PL, phospholipid. Flagella are not illustrated. The secondary wall
polymer is frequently teichoic acid. Reproduced with permission from proteins; however, there are fundamental differences in the
Hancock I and Poxton I (1988) Bacterial Cell Surface Techniques. London: sequence of the RNA and in the proteins that form the
John Wiley & Sons. complete ribosome. These differences are reflected in the

ENCYCLOPEDIA OF LIFE SCIENCES / & 2001 Nature Publishing Group / www.els.net 3

 Antibacterial Agents: Basis of Action
sedimentation properties of the ribosomes when subjected
to ultracentrifugation: bacterial ribosomes have a sedi-
mentation coefficient of 70 Svedberg units (70S), composed
of 50S and 30S subunits; the corresponding values for
mammalian ribosomes are 80S, with 60S and 40S subunits.
Bacterial chromosome
Although the genetic information of all living cells resides
in the double helix of deoxyribonucleic acid (DNA), the
organization of the bacterial chromosome is quite different
from its mammalian counterpart. Thus the bacterial
chromosome exists as a covalently linked circular mole-
cule, which in E. coli is about 1300 mm long. The bacterium
solves the problem of packaging this enormous molecule
into a cell that is only about 2 mm in length by condensing
the double helix down and then introducing further
constraints to produce a so-called ‘supercoiled’ state. This
structure has to be replicated every 20 min at the fastest
growth rates and must allow the transcription of messenger
RNA (mRNA) to provide all the necessary material for the
ribosomal workbench. Specialized enzymes, topoisome-
rases, perform the remarkable functions of nicking,
unwinding and resealing the double helix during replica-
tion and transcription and, although similar enzymes exist
in the mammalian cell, those in bacteria are sufficiently
different to be the basis of selective inhibitors such as the
antibacterial quinolones.
Metabolic transformations
Few antibacterial agents interfere directly with the
numerous metabolic processes taking place in the bacterial
cell cytoplasm, although the folic acid pathway has proved
amenable to selective attack by sulfonamides and diami-
nopyrimidines. These compounds exploit the requirement
of bacteria to manufacture folic acid de novo or differences
in affinity for dihydrofolate reductase (see below). Tetra-
hydrofolic acid is involved in various one-carbon transfers
within cells and its inhibition interferes with nucleic acid
synthesis, among other things.
Inhibitors of Bacterial Cell Wall
Synthesis
Several stages in the formation of the bacterial cell wall
provide targets for therapeutically useful antibacterial
agents (Figure 4). The first step is the formation of N-
acetylmuramic acid (NAMA) by the condensation of
phosphoenolpyruvate with N-acetylglucosamine (NAG),
a reaction which is inhibited by the phosphonic acid
antibiotic, fosfomycin. NAMA is next substituted by five
amino acids, the last two of which, d-alanyl-d-alanine (d-
ala-d-ala), is added as a unit. d-alanine is derived from l-
4 ENCYCLOPEDIA OF LIFE SCIENCES / & 2001 Nature Publishing Group / www.els.net
alanine by alanine racemase and the dipeptide is formed by
a ligase; both reactions are competitively inhibited by
cycloserine, a second-line antituberculosis agent. Any
amino acids needed for interpeptide bridges are now added
and the cell wall unit is completed by the addition of NAG
to the NAMA-peptide.
All this takes place in the cell cytoplasm, and the cell wall
unit has to be transported across the cytoplasmic
membrane to the peptidoglycan growth site. This is
accomplished through the agency of a lipid (55-carbon
isoprenyl phosphate) carrier molecule in the membrane. In
this process the lipid acquires an additional phosphate
group that is then removed to regenerate the carrier
function. This dephosphorylation reaction is inhibited by
the cyclic peptide, bacitracin, an antibiotic that is too toxic
for systemic use but is used in topical preparations.
The cell wall unit is now transferred to the growing end
of the peptidoglycan chain and this process is prevented by
glycopeptide antibiotics, such as vancomycin and teico-
planin. Finally, adjacent peptidoglycan chains are cross-
linked to give the wall its mechanical strength, and this
transpeptidation reaction is the site of action of penicillins
and other b-lactam antibiotics. However, the precise action
of these compounds is more complex
Mode of action of b-lactam agents
Although all b-lactam antibiotics interfere with the cross-
linking reaction that gives the bacterial cell wall its shape
and strength, different members of the b-lactam family
achieve this in subtly different ways. Growth and division
of the bacterial cell requires different forms of wall growth,
depending on whether the cell is engaged in extending the
cylinder of rod-shaped cells, forming the poles of the
dividing cell, or laying down the septum that will form the
division site. A battery of enzymes with transpeptidase,
endopeptidase and carboxypeptidase activity is required to
carry out these coordinated activities, and b-lactam
antibiotics can interact differentially with a number of
them, collectively called ‘penicillin-binding proteins’
(PBPs). In E. coli there are at least seven such PBPs,
numbered in order of decreasing molecular weight. Three,
the PBP-1 complex, PBP-2 and PBP-3 are involved in the
antibacterial action of b-lactam antibiotics. Most b-lactam
agents bind to all these PBPs, although at different
concentrations. In sufficient concentration they cause
rapid lysis of susceptible Gram-negative bacteria as the
defective cell wall fails to protect the cell from osmotic
rupture. Others, including cephalexin, cephradine and
aztreonam, selectively bind to PBP-3, which is involved in
septum formation, and allow the cells to continue to grow
into long multinucleate filaments before death ensues.
Some others, including mecillinam (amdinocillin), imipe-
nem and the b-lactamase inhibitor, clavulanic acid,
preferentially bind to PBP-2. This PBP is involved in
 Antibacterial Agents: Basis of Action

L-ala ×2

D-ala ×2 Cycloserine

Amino acids D-ala-D-ala

β-Lactam
NAMA NAMA-pentapeptide Glycopeptides antibiotics
Fosfomycin + Lipid carrier Transfer to Crosslinking
(membrane) peptidoglygan
NAG NAG
Bacitracin

Dephosphorylation
of lipid

Figure 4 Sites of action of inhibitors of bacterial cell wall syntheses. NAMA, N-acetylmuramic acid; NAG, N-acetylglucosamine. Reproduced with
permission from Greenwood D, Slack RCB and Peutherer JF (eds) (1997) Medical Microbiology, 15th edn. Edinburgh: Churchill Livingstone.

extension of the cell wall cylinder, and affected cells
gradually round up before succumbing to osmotic lysis.
Gram-positive bacteria also have multiple PBPs, but
fewer than are found in Gram-negative rods. Inhibition of
the PBPs causes a release of lipoteichoic acid, and this
triggers an autolytic dismantling of the peptidoglycan.
Because the cell wall peptidoglycan is much thicker, death
occurs more slowly. In the case of ‘methicillin-resistant
Staphylococcus aureus’, which are resistant to all b-lactam
antibiotics, the bacteria produce a novel PBP (PBP-2’) that
has a low affinity for b-lactam compounds and allows the
cells to continue manufacturing intact cell wall.
Inhibitors of Bacterial Protein Synthesis
Various clinically useful antibacterial agents inhibit
protein synthesis by directly inhibiting the functions of
ribosomes, the cellular organelles upon which the genetic
message is decoded to produce proteins. The specific action
of antibacterial protein synthesis inhibitors arises from
differences in the structure of bacterial and mammalian
ribosomes (see Bacterial Structure, above) allowing unique
binding sites in the bacterial ribosome to be selectively
exploited by the antibacterial agents. In some cases
differential accumulation by bacterial and mammalian
cells also contributes to the selective action of drugs
inhibiting protein synthesis; for example, the tetracyclines
and fusidic acid are more readily accumulated by bacteria
than by mammalian cells.
Bacterial protein synthesis is a complex process invol-
ving the formation of active 70S particles capable of
catalysing peptide bonds between incoming amino acids in
a repetitive process known as the elongation cycle (Figure 5).
Most clinically useful inhibitors of bacterial protein
synthesis, including lincosamides, macrolides, tetracy-
clines, chloramphenicol, aminoglycosides, streptogramins
and fusidic acid, act by inhibiting or modifying this cycle
(Figure 5).
Each of these drugs binds to separate sites in the 70S
ribosomal particle and affects different stages of the
elongation cycle: lincosamides and chloramphenicol in-
hibit peptidyl transferase activity; the macrolides cause
dissociation of peptidyl tRNA (transfer RNA) from the
ribosome; the tetracyclines inhibit the binding of aminoa-
cyl tRNA; streptogramins inhibit both the binding of
aminoacyl tRNA and peptidyl transferase activity; and
fusidic acid forms a stable complex between the ribosome
and the soluble elongation factor G (EF-G), the release and
reassociation of which are important events that facilitate
the elongation cycle. Aminoglycosides have a variety of
effects, including prevention of the movement between
ribosomes and mRNA (spectinomycin), misreading of
proteins being translated (streptomycin), and inhibition of
binding of EF-G to the ribosome (neomycins, gentamicins,
kanamycins, amikacin and tobramycin).
In contrast to most protein synthesis inhibitors, which
affect the elongation cycle, linezolid, a member of a new
class of antibacterial agents, the oxazolidinones, prevents
the formation of the 70S initiation complex by binding to
the 50S ribosomal subunit (Figure 5).
Inhibitors of Nucleic Acid Synthesis
Synthesis of RNA and DNA are essential processes in the
bacterial cell and can be selectively disrupted by anti-
bacterial agents.

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 Antibacterial Agents: Basis of Action

Initiation factors
Initiator formyl-methionine-tRNA
mRNA
AUG fMet

Preinitiation
Peptide complex
product Linezolid
30S subunit

mRNA + AUG GCU CGC

Formation of
Acceptor 70S initiation
Peptidyl (A) site complex
(P) site
Fusidic acid fMet
Neomycin 50S subunit
Tetracycline
Gentamicin Streptogramins
Kanamycin Elongation cycle involving
Amikacin accurate reading of genetic code and association of peptidyl-tRNA with P site
Tobramycin Delivery of charged
Streptomycin Macrolides tRNA, e.g. Ala-tRNA
Termination ala by elongation factor
Tu
Translocation of
growing polypeptide
AUG GCU CGC chain from A to P site AUG GCU CGC AUG GCU CGC
Empty driven by elongation
A site factor G
Peptide bond
ala Movement between ala formation by fMet ala

mRNA and ribosome peptidyl transferase

Growing fMet fMet

polypeptide Spectinomycin
chain Chloramphenicol
Lincosamides
Streptogramins

Figure 5 Bacterial protein synthesis, showing the steps that are inhibited by various antibacterial agents.

Inhibition of RNA synthesis
RNA polymerases are enzymes that mediate transcription
of structural genes by catalysing the initiation and
elongation of RNA molecules on a DNA template.
Bacterial RNA polymerases comprise a core enzyme (E)
and a specificity subunit, also defined as a sigma factor (s).
Core polymerase (E) is a stable, noncovalent assembly of
four polypeptide chains consisting of two a subunits (each
with a molecular weight of 36 500 Da), one b subunit
(molecular weight 151 000 Da) and one b’ subunit (mole-
cular weight 155 000 Da). Under normal growth condi-
tions transcription of most genes is initiated by RNA
polymerase holoenzyme Es70 in which the specificity
subunit is s70. Under other growth conditions, e.g. during
the stationary phase of growth, alternative sigma factors
are used to achieve expression of a different set of genes.
Rifampicin, a member of the rifamycin group of
antibiotics, binds selectively to the b subunit of RNA
polymerase and in so doing interferes with the ability of the
holoenzyme (Es) to initiate RNA synthesis. Binding of
RNA polymerase to the DNA template is not blocked and
inhibition probably results from interference with the
formation of the first phosphodiester bond in the RNA
chain. The selective antibacterial action of rifampicin
reflects structural differences between the RNA poly-
merases of bacterial and mammalian cells. In contrast to
the bacterial enzymes, mammalian RNA polymerases
contain 10 or more polypeptide chains, none of which
contain sites for the binding of rifampicin.
Inhibition of DNA synthesis
DNA gyrase inhibitors
Replication of the bacterial chromosome, a circular duplex
DNA molecule, requires the separation of the two highly
intertwined parental strands. However, separation of
strands wound in a helix generates loops, termed positive
supercoiled twists, in the single strands. Unless prevented,
this situation would stop further unwinding of parental
DNA at the replication fork. The enzyme DNA gyrase (a
type II DNA topoisomerase) relaxes positively supercoiled
DNA by periodically breaking a phosphodiester bond in
one of the strands of the double helix, introducing negative
supercoils, and finally resealing the nick.
Bacterial DNA gyrase is a tetrameric enzyme composed
of four subunits: two gyrase A subunits (each with a
molecular weight of 97 000 Da) and two gyrase B subunits
(each with a molecular weight of 90 000 Da). All activities

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of the enzyme require both A and B subunits, but certain
domains mediate different functions. The A subunits of
gyrase are involved in the DNA breakage and resealing
events associated with supercoiling, while the B subunits
are responsible for adenosine triphosphate (ATP) hydro-
lysis, reflecting the consumption of energy in the formation
of negative supercoils.
Bacteria contain a second type II DNA topoisomerase
called DNA topoisomerase IV. This is a heterodimer with
subunits of molecular weight 70 000 and 75 000 Da. DNA
topoisomerase IV catalyses ATP-dependent relaxation of
negatively and positively supercoiled DNA. However,
unlike gyrase, topoisomerase IV displays no DNA super-
coiling activity and appears to have a primary role in
chromosomal partitioning (segregation) to daughter cells
during cell division.
The quinolone antibacterial agents, including nalidixic
acid and the newer fluoroquinolones, selectively inhibit
DNA gyrase and topoisomerase IV and therefore arrest the
essential process of bacterial DNA replication. In Gram-
negative bacteria the primary target of quinolones is DNA
gyrase, whereas in Gram-positive bacteria the primary
target is DNA topisomerase IV. These drugs do not bind
directly to DNA gyrase or topoisomerase IV, but form
stable drug–DNA–enzyme complexes that prevent the
further functioning of the topoisomerases.
Agents mediating strand breakage
Nitroimidazoles, such as metronidazole and tinidazole,
interfere with DNA synthesis by introducing strand
breakages in duplex DNA. These drugs kill bacteria, and
some protozoa, that inhabit a low redox environment and
are thus capable of anaerobic metabolism. The activity of
these agents depends upon reduction, within the cell, of the
nitro group that each drug contains. Reduction results in
the formation of a number of products including the nitro
radical anion, which reacts with DNA, oxidizing it and
causing strand breaks and destabilization of the double
helix. Nitrofurans, like nitrofurantoin, probably act in an
analogous way, albeit in an aerobic environment.
Antifolate Compounds
Sulfonamides are structural analogues of p-aminobenzoic
acid. They interfere with the early stages of folic acid
synthesis by competitive inhibition of dihydropteroic acid
synthetase, which condenses p-aminobenzoic acid with
dihydropteroic acid. The sulfonamide may also be
erroneously incorporated into the folic acid molecule to
produce an inactive product. Bacterial cells synthesize folic
acid, whereas mammalian cells use the preformed dietary
vitamin, and this is the basis of the selective antibacterial
action of sulfonamides.
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Antibacterial Agents: Basis of Action
Diaminopyrimidines, like trimethoprim and the anti-
malarial compound, pyrimethamine, act at a later stage on
the same pathway by inhibiting dihydrofolate reductase,
the enzyme that generates the active product, tetrahydro-
folate, from dihydrofolate. The affinity of trimethoprim
for dihydrofolate reductase of bacteria is several orders of
magnitude higher than the affinity for the mammalian
enzyme; similarly pyrimethamine has a very high affinity
for the dihydrofolate reductase of malaria parasites.
Because sulfonamides and diaminopyrimidines act on
the same metabolic pathway, they exhibit a strongly
synergic interaction, at least in vitro. However, because
tetrahydrofolate is reoxidized to dihydrofolate during the
biosynthesis of thymidylic acid, diaminopyrimidines ra-
pidly trap the vitamin in the unusable dihydrofolate form.
Sulfonamides, in contrast, cut off the supply of dihydro-
folate and act rather slowly because the folate pool
becomes depleted only after several cell divisions. For this
reason, if there is sufficient diaminopyrimidine present to
halt tetrahydrofolate regeneration completely, the sulfo-
namide does not have an opportunity to contribute to the
antibacterial action.
Membrane Disorganizing Agents
The structurally related cyclic antibiotics polymyxin B and
polymyxin E (colistin), which are primarily active against
Gram-negative bacteria, exert their antibacterial activity
by disrupting both the outer and inner (cytoplasmic)
membranes of the Gram-negative cell. Death of the cell
results from leakage of cytoplasmic contents. The prefer-
ential action of polymyxins on Gram-negative bacteria
results in part from their targeting of lipopolysaccharide,
found exclusively in the outer membrane of the Gram-
negative cell, and their preference for cytoplasmic mem-
branes containing the phospholipid phosphatidylethano-
lamine, which is present in Gram-negative bacteria, but is
generally lacking in Gram-positive species. Since the
polymyxins also interact with mammalian cell membranes
and exhibit a number of adverse side effects, they have only
a minor place in medicine.
Among other agents that act on bacterial membranes are
the endogenous host defence peptides of animals, such as
defensins, magainins and cecropins. These are cationic
oligopeptides that appear to form channels in the bacterial
cytoplasmic membrane, leading to disruption of the
transmembrane proton gradient and death of the cell.
Selectivity for bacteria appears to result from a number of
factors, including the absence of cholesterol in bacterial
membranes.
7
 Antibacterial Agents: Basis of Action
Antimycobacterial Drugs
Several antibiotics that have already been described have
roles in the treatment of infections caused by mycobacter-
ia, such as tuberculosis and leprosy. These include
cycloserine, some fluoroquinolones and macrolides, ri-
fampicin and streptomycin. These agents inhibit the same
targets in mycobacteria as they do in other bacteria.
Drugs that are used only in the treatment of mycobac-
terial infections appear to interact with targets that occur
exclusively in these organisms. The antituberculosis
agents, isoniazid and ethionamide, are oxidized within
the mycobacterial cell, and one or more of the oxidation
products interact with enzymes involved in the biosynth-
esis of mycolic acids, unique and essential components of
the mycobacterial envelope. Ethambutol, which also has a
role in the treatment of tuberculosis, inhibits the biosynth-
esis of arabinogalactan, another essential macromolecule
in the mycobacterial wall. The molecular modes of action
of dapsone and clofazimine (important antileprosy drugs)
and pyrazinamide (an antituberculosis agent) are un-
known.
Future Directions
The development of effective chemotherapy for bacterial
infections represents one of the most remarkable achieve-
ments of the twentieth century. Unfortunately, the
emergence of resistance to existing antimicrobial agents
increasingly threatens their effectiveness for the treatment
of certain infections, and the discovery of new drugs is now
a matter of urgency. In order to minimize the potential for
crossresistance to existing drugs, new drugs should ideally
be aimed at previously unexploited biochemical targets in
the bacterial cell. A good example concerns the potential
8 ENCYCLOPEDIA OF LIFE SCIENCES / & 2001 Nature Publishing Group / www.els.net
for developing drugs that would inhibit lipopolysaccharide
synthesis in Gram-negative bacteria. The lipid A compo-
nent of lipopolysaccharide anchors the molecule in the
outer membrane, and mutants deficient in the synthesis of
lipid A are nonviable. Since several of the enzymes involved
in lipid A synthesis are unique to Gram-negative bacteria,
the opportunity for selectivity of drug action exists.
Much interest has also been shown in derivatives of the
naturally occurring cationic peptides that disrupt bacterial
membranes in a unique way, and research continues to seek
other means to subvert or nullify the virulence of
pathogenic bacteria.
Whether these developments come to fruition as
clinically useful agents remains to be seen, but the twin
challenges to husband the resources we have painstakingly
acquired, and to maintain the impetus of discovery in the
future, must surely be met.
Further Reading
Chopra I and Brennan P (1998) Molecular action of anti-mycobacterial
agents. Tubercle and Lung Disease 78: 89–98.
Greenwood D (ed.) (1995) Antimicrobial Chemotherapy, 3rd edn.
Oxford: Oxford University Press.
Hancock REW and Lehrer R (1998) Cationic peptides: a new source of
antibiotics. Trends in Biotechnology 16: 82–88.
O’Grady F, Lambert HP, Finch RG and Greenwood D (eds) (1997)
Antibiotic and Chemotherapy, 7th edn. Edinburgh: Churchill Living-
stone.
Pratt WB and Fekety R (1986) The Antimicrobial Drugs. Oxford: Oxford
University Press.
Russell AD and Chopra I (1996) Understanding Antibacterial Action and
Resistance, 2nd edn. London: Ellis Horwood.
Williams RAD, Lambert PA and Singleton P (1996) Antimicrobial Drug
Action. Oxford: Bios Scientific Publishers.
Wyckoff TJO, Raetz CRH and Jackman JE (1998) Antibacterial and
anti-inflammatory agents that target endotoxin. Trends in Microbiol-
ogy 6: 154–159.