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International Journal of Antimicrobial Agents xxx (2013) xxx–xxx

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International Journal of Antimicrobial Agents

journal homepage: http://www.elsevier.com/locate/ijantimicag

Review

Alternative natural sources for a new generation of antibacterial

agents
Peter W. Taylor ∗
University College London School of Pharmacy, 29–39 Brunswick Square, London WC1N 1AX, UK

a r t i c l e i n f o a b s t r a c t

Article history: The huge challenge posed by antibiotic resistance would be well served by the discovery and development
Received 13 May 2013 of a new wave of antibacterial drugs. Natural products have been the mainstay of anti-infective drug dis-
Accepted 13 May 2013 covery since the early days of the antibiotic era, but mining of valuable natural resources has been all but
abandoned by the major pharmaceutical players in favour of synthetic chemistry. The search for naturally
Keywords: occurring antibacterial agents has continued in academe, but activities need to be repositioned to take
Secondary metabolites
advantage of exciting advances in genomics and advanced genetic engineering. This review evaluates
Complex microbial communities
the potential of microbial communities in underexplored environmental niches to yield new antibiotics
Plant-derived agents
Membrane-interactive agents
and the harnessing of biomembrane-interactive plant-derived agents as supplements to conventional
Phenotype modification antibacterial chemotherapy.
© 2013 Published by Elsevier B.V.

1. Introduction Hopes that exploitation of genomics, in silico drug design and

Naturally occurring molecules, predominantly secondary
metabolites, are the source of the majority of drugs in clinical
use today. This is especially true of anti-infective agents; the
most recent comprehensive survey of drug source by Newman
and Cragg [1] indicated that natural products, or drugs based on
natural product scaffolds, account for >75% of the 97 approved
antibacterial New Chemical Entities introduced over the period
1981–2006, highlighting the continuing importance of natural
products as the basis of new therapeutics for bacterial infectious
diseases. Naturally occurring antibacterial agents developed for
clinical use have, without exception, been sourced from micro-
organisms, with the overwhelming majority derived from moulds
and members of the actinomycetes, notably the genus Strepto-
myces [2]. It has been estimated [3] that this bacterial genus alone
elaborates ca. 100 000 secondary metabolites with at least some
antimicrobial activity, suggesting that continuation of traditional
antibacterial screening strategies would be likely to unearth new
compounds with mechanisms of action distinct from those of
current, clinically relevant antibiotics [1]. However, commercial
screening of microbial products for novel drug candidates has
fallen from favour in recent years: the intensive application of
traditional screening methods invariably yielded more ‘old friends’
than novel compounds [3] as discovery programmes progressed.
∗ Tel.: +44 20 7753 5867; fax: +44 20 7753 5867.
E-mail address: peter.taylor@ucl.ac.uk
target-based high-throughput screening of combinatorial com-
pound libraries would yield a new generation of novel drugs
addressing new molecular targets have not been realised despite
massive investment [4,5]. For these reasons, there is growing
interest in the potential of untapped microbial, plant and animal
sources to yield novel molecules, scaffolds and pharmacophores
for a new generation of antibacterial drugs [6,7].
Although major pharmaceutical concerns currently devote
much less resource to antibacterial drug discovery programmes
compared with their investment during the ‘golden age’ of dis-
covery, they continue to uncover novel structures with significant
antibiotic activity. For example, Bayer AG discovered, but did
not develop, novel antibiotics of the acyldepsipeptide class that
bind to a bacterial protein required for regulation of cellular
proteolytic activity [8]. In a similar fashion, Merck Research Labora-
tories unearthed the previously identified antibiotic platensimycin,
an inhibitor of bacterial type II fatty acid synthesis, from an
established collection of natural product extracts [9]. Recent tech-
nological advances in conventional screening strategies that filter
out rediscoveries, allow synthetic tailoring of key natural prod-
uct scaffolds and facilitate repurposing of compound libraries from
non-antibiotic discovery programmes have rekindled interest in
commercial high-throughput searches for new natural product
scaffolds from micro-organisms, with an emphasis on the mining
of underexplored ecological niches [10]. In addition, plants [11],
social insects [6,12] and higher animals [13] are attracting atten-
tion as potential sources of antibacterial therapeutic agents; in this
context, mammalian cationic amphiphilic peptides possess potent
antimicrobial and immunomodulatory activities that together may

0924-8579/$ – see front matter © 2013 Published by Elsevier B.V.

http://dx.doi.org/10.1016/j.ijantimicag.2013.05.004

Please cite this article in press as: Taylor PW. Alternative natural sources for a new generation of antibacterial agents. Int J Antimicrob Agents
(2013), http://dx.doi.org/10.1016/j.ijantimicag.2013.05.004
 ARTICLE IN PRESS
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2 P.W. Taylor / International Journal of Antimicrobial Agents xxx (2013) xxx–xxx
represent a viable alternative approach to the treatment of bacterial
infections [13,14]. This review will examine the potential of some
of these alternative sources of antibacterial compounds to act as
lead structures for a new generation of antibiotic agents.
2. The lure of biodiversity
The large majority of clinically useful antibiotics were discov-
ered as secondary metabolites of bacteria resident in the soil.
Selman Waksman’s systematic studies of the production of antibac-
terial chemical substances by soil micro-organisms in the years
following the Second World War led to the discovery of strepto-
mycin and highlighted the capacity of complex environments to
yield microbes with the potential to elaborate novel molecules with
antibiotic activity. The focus on diverse microbial communities in
soil has been at least partly driven by the perception that produc-
tion of antibiotic substances gives some microbes a competitive
advantage in a hostile environment, although it has recently been
emphasised that microbial communities are also highly commu-
nicative and the primary role of apparent antibiotic substances may
be to effect microbial cross-talk in order to maintain, rather than
perturb, complex microbial populations [15]. Soils are rich sources
of microbial biodiversity, with up to 5000 bacterial species repre-
sented in 1 g of soil typically containing ca. 109 bacteria [16], many
of which elaborate chemical structures of pharmaceutical interest.
There are, however, other biodiverse and complex microbial
communities that have not yet been mined to any extent for novel
anti-infective compounds: these include lakes, rivers, sediments
and marine environments and even niches within mammals such
as the gastrointestinal tract and saliva [15]. Micro-organisms that
occupy sites characterised by extremes of temperature, pressure
and light availability, such as polar sea ice [17] and ice cores
[18], hydrothermal vents [19,20], sub-seafloor sites [21] and caves
[22,23], represent potentially rich sources of novel compounds by
virtue of highly specialised metabolic adaptations. Recent advances
in marine metagenomics have revealed the complexity, diversity
and high variability of the microbial communities in the seas and
oceans that cover 70% of the Earth’s surface. Approximately 60% of
the ocean floor is covered by water more than 2000 m deep, and
emerging insights into the biogeographical patterns of bacterial
communities, reflecting physical, chemical and biological contrasts
and interconnections between the pelagic and the benthic realms
[24], have emphasised the potential of the estimated 1029 individ-
ual marine bacterial and archeal cells [25] as repositories of novel
chemical structures. This biodiversity has been exploited by niche
pharmaceutical companies and institutions as a source of anti-
cancer, antiviral and antipsychotic drugs, either approved or under
clinical development [26], but marine micro-organisms remain an
underutilised source of novel anti-infective agents. The degree of
biodiversity of marine prokaryotes does not rival that of soil [25],
but the sheer numbers of potential ‘microbial factories’ involved,
the availability of large culture collections of marine fungi, bacte-
ria and Archaea as well as significant compound libraries of marine
natural products [27] would support a concerted effort to investi-
gate these resources for novel lead compounds.
In contrast to these emerging microbial sources, there has been
an enormous effort over a considerable period of time to exploit
the plant kingdom as an alternative source of antibacterial lead
molecules, with notable lack of success to date [11]. The popularity
of plant materials as the starting point for the discovery and devel-
opment of compounds with activity against bacterial pathogens
shows few signs of waning: major international conferences for
ethnobiology and ethnopharmacology feature large numbers of
presentations on the in vitro antibacterial activity of plant extracts
and purified plant metabolites and there is a huge literature base
in this area. The reasons for this are straightforward: there is
Please cite this article in press as: Taylor PW. Alternative natural sources for a new generation of antibacterial agents. Int J Antimicrob Agents
(2013), http://dx.doi.org/10.1016/j.ijantimicag.2013.05.004
enormous biodiversity within the plant kingdom, with the number
of species of flowering plants estimated to be in excess of 300 000
[28]; plants elaborate a large number of low-molecular-mass sec-
ondary metabolites, estimated to be in the region of 100 000 distinct
molecular species [29]; plant-derived medicines have been used for
millennia in many parts of the world to treat a wide variety of con-
ditions, including infections [30], and it is assumed that individuals
would not continue to do so if they derived no benefit; some valu-
able ethical drugs for the treatment of parasitic infections have been
developed from plants, such as the antimalarial agent artemisinin
[31], obtained from a plant used in China for almost 2000 years;
and, issues of biodiversity loss notwithstanding [32], plant mate-
rials are usually readily available. Are there factors holding back
the discovery and translation to medical practice of plant-derived
antibacterial compounds?
3. Pros and cons of plant-based antibacterial agents
The majority of the world’s population depends on traditional
medicine for their health, and local providers are consulted owing
to perceptions of a positive outcome [33]. Traditional medicines
are often employed alongside biomedicines so their continued use
is not due primarily to a lack of access to modern healthcare; lack
of sufficient recognition by biomedical practitioners of the value
and importance of indigenous medical systems may play a part in
their retention by native communities [34]. There is no established
research community for the evaluation of traditional plant-based
medicines, particularly in countries where their use is common,
and any intrinsic value of plant extracts as therapeutic concoc-
tions is not based on modern perceptions of efficacy and safety as
determined by controlled clinical trials. Therapeutic use of poorly
evaluated, complex and ill-defined extracts of variable composi-
tion is at odds with the principles of modern medicine. In fact, the
increasing interest and acceptance of complementary and alter-
native medicine in Western societies is causing concern regarding
doubts over efficacy and safety [35]. With encouragement from the
World Health Organisation (WHO) [36], there have been a number
of recent attempts to investigate the safety, efficacy and quality of
traditional medicines using evidence-based approaches. Very little
evidence has been obtained for any significant degree of efficacy for
the large majority of traditional medicines examined (for exam-
ple, [37,38]), they may disadvantageously interact with modern
medicines [39] and may do harm [40]. Only a very limited num-
ber of herbal remedies for the treatment of infections have been
subjected to evidence-based evaluation. For example, Umckaloabo
is an extract of Pelargonium sidoides that is sold in Western pharma-
cies for the relief of acute respiratory infections; doubts have been
expressed, following critical evaluation, over its claimed capacity to
alleviate the symptoms of acute rhinosinusitis, sinusitis and com-
mon cold in adults and acute bronchitis in adults and children [41].
More detailed evaluation of the antimicrobial efficacy of traditional
medicines clearly needs to be undertaken before such concoctions
based on plant extracts can be considered as a starting point for the
purification of bioactive chemical constituents for development as
modern medicines.
The body of data recording the in vitro antibacterial activ-
ity of plant extracts and constituents is very large and indicates
that activity as determined by the minimum inhibitory concen-
tration (MIC) is generally weak. A survey of publications in this
area for 2012–2013 shows that a considerable number of stud-
ies have utilised unfractionated extracts of root and aerial parts
of various plants, with MIC values for crude extracts generally in
excess of 100 mg/L, and often in the mg/mL range, against common
pathogens such as Staphylococcus aureus, Klebsiella pneumoniae and
Pseudomonas aeruginosa [42–44]. It is difficult to envisage how
such materials could be developed as potential sources of therapies
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for the infections indicated by the authors: an effective antibiotic of low-molecular-mass markers from modelled membranes com-
should have high selective toxicity, with potent activity at achiev- posed of Escherichia coli phospholipids [58]. The major phenolic
able concentrations against target bacteria and minimal effects constituent of the olive leaf, oleuropein, showed significant lev-
on the host, chemical and physical stability after formulation to els of partition into phospholipid membranes, adopting a location
comply with the pharmaceutical specification, good bioavailability close to the membrane surface and causing perturbations that are
when administered by the chosen route, an appropriate phar- likely to account for its antimicrobial activity [59]. The main bioac-
macokinetic profile and, ideally, not readily give rise to resistant tive components of the African potato, Hypoxis rooperi, inhibited the
variants. growth of S. aureus and, to a lesser extent, E. coli, concomitant with
The extensive array of plant-derived compounds with antimi- the induction of membrane leakage following shallow interactions
crobial activity has been the subject of major reviews [45–47], with the phospholipid components [60]. The polyphenolic cate-
and structures, sources and potency will not be reviewed here chins and catechin gallates have very weak activity (MICs of 64 mg/L
in any depth. These aromatic secondary metabolites belong to to >256 mg/L) against meticillin-resistant S. aureus (MRSA) strains
a relatively small number of chemical categories and, where and other Gram-positive bacteria [61]; they bind to, and penetrate,
function is known, participate in plant defence and impart pig- model bilayers [62] and staphylococcal membranes [52] to differ-
ments and odours. Alkaloids, such as berberine, have long been ing extents, reflecting their capacity to modulate the biophysical
known to have antiparasitic activity, but some of these hetero- properties of these bilayers. It would be interesting to extend these
cyclic nitrogenous compounds have more recently been shown studies to include other weakly bioactive phytochemicals.
to possess moderate-to-weak (MICs of 8–64 mg/L) activity against
rapidly growing mycobacteria and staphylococci [48]. Coumarins
4. Induction of protective phytochemicals by microbial
are phenolic molecules composed of fused benzene and ␣-pyrone
infection
rings and are widely distributed within the plant kingdom; the
established coumarin antibiotic novobiocin, produced by some
The discovery of novel antibacterial agents in solvent extracts
Streptomyces spp., has potent activity against Gram-positive bacte-
of plants most frequently begins with leaves or roots from healthy
ria. Plant-derived coumarins are ineffective against Gram-negative
specimens, even though there is ample evidence that many key
bacteria but show good activity against some Gram-positive
components of plant defences against phytopathogens are induced
species. They are inhibitors of bacterial DNA gyrase [49]. Flavonoids
by infection. In fact, plants respond to microbial attack through
and isoflavonoids are widely distributed phenolics containing one
a highly co-ordinated repertoire of molecular, cellular and tissue-
carbonyl group and constitute a major group of plant metabolites.
based defensive barriers to colonisation and invasion [63,64]. There
As well as antioxidant properties that have evolved to protect plants
is a strong case for the evaluation of induced components of the
from environmental stresses, these compounds display weakly
defensive repertoire, as they are produced selectively in response
antibacterial activities [50], although the propensity of some mem-
to invasion by specific plant pathogens. Formidable constitutive
bers of this chemical class to intercalate into biomembranes enables
defences include preformed barriers to invasion such as cell walls,
them to favourably modify the properties of at least some Gram-
waxy epidermal cuticles and bark; detection of invading pathogens
positive pathogens [51,52]. The capacity of catechin gallates to
leads to an inducible response that includes the elaboration of
induce novel phenotypes in this way will be dealt with in another
toxic chemicals and depolymerases, as well as apoptosis. Some
section of this review. The highly reactive and ubiquitous quinones
chemical defence components, such as the terpenoid essential oils,
are aromatic compounds with two ketone substitutions. Members
are produced constitutively, as they function as insect toxins and
of the class have weak antibacterial activity [53,54] that can be
are relatively harmless to humans [65], but the concentration and
improved by rational design [55] but they possess significant mam-
range of chemicals produced in response to perceived attack from
malian toxicity [56]. Terpenoids are isoprene-based constituents
opportunistic micro-organisms increase enormously after detec-
of essential oils and are the largest group of natural products
tion of the invaders [63]. There is a substantial literature on the
[57]. Some members of this class of molecules possess good-to-
capacity of plant-associated bacteria as agents for the management
moderate activity against Gram-positive bacteria [46], although
of soil and plant health [66] but virtually no published accounts of
very few of the ca. 25 000 terpenoid structures reported have been
the use of pathogen-stimulated plants as a potential source of novel
evaluated for bioactivity. Some have significant toxicity following
antibacterial compounds, although Lewis and Ausubel [11] indi-
ingestion by mammals [57]. Tannins, lignans, steroidal saponins
cate that such screening has taken place in industrial laboratories.
and xanthones have also been shown to possess weak-to-moderate
Most, if not all, plants respond to fungal and bacterial pathogens
antibacterial activity [45,47].
by activation of low-molecular-weight antimicrobial compounds
With the exception of a small number of coumarins, very little is
[67], many of which differ from the constitutively expressed palette
known of the antibacterial mechanism of action of these molecules.
of antibacterial agents described above. These ‘phytoalexins’ rep-
Some weakly antibacterial compounds afford protection only when
resent a source of antibacterial agents and the complexity of the
synthesised by the host plant in large quantities; they resemble
response, involving a wide array of chemical structures, coupled
weak antiseptics and are unlikely to act through a target-specific
with significant variation depending on the nature of the pathogen
mechanism in a ‘lock-and-key’ fashion [11]. Further, Lewis and
and plant type [67,68], could provide a rich resource for drug dis-
Ausubel [11] have reported that a number of major pharmaceu-
covery. The plant response to infection can also be induced by plant
tical companies and smaller biotech concerns undertook extensive
activators, such as analogues of salicylic acid [69] and imprimatins
screening of plant materials for potent, non-toxic, broad-spectrum
[70], greatly simplifying the experimental design of activator induc-
antibiotics and failed to find suitable lead structures or devel-
tion processes.
opment candidates. In support of a relatively non-specific mode
of action for many phytochemicals, Micol and co-workers have
determined that some phenolics, polyphenols and quinones alter 5. Disarming bacterial pathogens with
the biophysical properties of membrane bilayers by intercalation membrane-interactive, plant-derived natural products
into, and sometimes perturbation of, the lipid palisade. Thus, the
1,8-dihydroxyanthraquinones barbaloin and emodin displayed a It has been cogently argued [11] that the failure of industry
high affinity for phospholipid membranes, weakened hydrophobic and academe to discover broad-spectrum plant-derived antibac-
interactions between hydrocarbon chains and engendered leakage terial compounds is likely to reflect the fact that plants have a

Please cite this article in press as: Taylor PW. Alternative natural sources for a new generation of antibacterial agents. Int J Antimicrob Agents
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chemical strategy for protection against microbial infection that

differs substantially from that employed by multicellular animals.
The affinity of some weakly antibacterial phenolics and polyphe-
nols for lipid bilayers and their capacity to induce varying degrees
of membrane perturbation [58–62,71,72] suggest that such com-
pounds may act in concert to provide a shield against microbial
attack. Similarly, the barberry plant, Berberis fremontii, synthesises
the multidrug resistance pump inhibitor 5 -methoxyhydnocarpin,
preventing extrusion from the bacterial cytoplasm of the alkaloid
DNA intercalator berberine [73].
Such modifying compounds could provide the basis for alter-
native modalities for the control of bacterial infections in a way
that would confound the rapid emergence of drug resistance. Mul-
tidrug efflux pumps make a significant contribution to intrinsic and
acquired antibiotic resistance of a wide range of clinically relevant
bacteria and there have been consolidated efforts, so far with-
out tangible success, to find clinically acceptable pump inhibitors
[74,75]. Inhibitors in combination with antibiotics whose action
is compromised by efflux pumps would be welcome additions
to the chemotherapeutic arsenal, even though the development
of combination therapies is notoriously difficult. These efforts
should be tempered by indications that pump inhibitors may
select multidrug-resistant mutants, at least in the laboratory [76].
A search for additional plant-derived multidrug resistance pump
inhibitors could provide a valuable resource for alternative antibac-
terial drug development programmes.
Fig. 1. Structure and space filling model of (−)-epicatechin gallate (ECg). The
The cytoplasmic membrane constitutes the selective barrier colourless regions are hydrophobic domains and the darker regions are hydrophilic
controlling cellular ingress and egress of materials and houses domains. Ring nomenclature assignment is shown for ECg.
many of the enzymes involved in bioenergetic functions, cell wall
synthesis and macromolecule secretion. Penetration of phyto-
chemicals into this phospholipid bilayer will have a profound effect
on a wide range of essential cell functions. In some cases, interca- isolates, inducing a reversible increase in susceptibility to oxacillin
lation of these chemical structures induces membrane disruption of the epidemic clone EMRSA-16 from 512 mg/L to 1 mg/L [61]; this
that permits solute equilibration across the bilayer, resulting in property was significantly enhanced by EC [79]. Galloyl catechins
cell death, but more subtle interactions at subinhibitory concen- also disrupt secretion of virulence-related proteins [82,83] and pre-
trations induce re-organisation of membrane architecture with vent formation of biofilms [84,85]. ECg compromises osmosensing
implications for the capacity of the target bacteria to cause dis- in halotolerant S. aureus by interfering with Na+ -specific antiporter
ease in the susceptible host. For example, the catechins possess systems operating across the cytoplasmic membrane [86] and pro-
negligible antibacterial activity against Gram-positive and Gram- motes cell wall thickening and cell aggregation without affecting
negative bacteria but are able to interact with, and penetrate, the rate or extent of growth [85].
lipid bilayers. These polyphenols constitute up to 30% of the dry The determinant of meticillin resistance in S. aureus is penicillin-
weight of the Japanese green tea (Camellia sinensis) leaf and are binding protein (PBP) 2a, encoded by the mecA gene initially
potent antioxidants; the most abundant constituents are the galloyl acquired from a related staphylococcal species; PBP2a is a cytoplas-
catechins (−)-epicatechin gallate (ECg) and (−)-epigallocatechin mic membrane-associated transpeptidase that is not susceptible
gallate (EGCg) and the non-galloyl molecules (−)-epicatechin (EC) to acylation by ␤-lactam agents [87] and functions co-operatively
and (−)-epigallocatechin (EGC) [51]. The galloyl catechins bind with PBP2 in the presence of high concentrations of ␤-lactams.
more avidly than EC or EGC to unilamellar vesicles [62,77,78] and Thus, MRSA continues to cross-link peptidoglycan chains in the
staphylococcal membranes [79] and penetrate deep within the presence of ␤-lactams even though the transpeptidase domain of
hydrophobic core of the lipid palisade [62]. Non-galloyl catechins PBP2 is inactivated [88]. ECg does not suppress the transcription of
such as EC adopt a superficial location within bilayers close to the mecA or the production of PBP2a and produces only small changes
phospholipid–water interface, but they elicit minor perturbations in the expression and functionality of other PBPs, resulting in a
to the membrane that are sufficient to enhance the access of ECg 5–10% reduction in peptidoglycan cross-linking without compro-
to sites deep within the hydrophobic core [79]. ECg (Fig. 1) has mising cell integrity [85]. ECg-mediated perturbation of orderly cell
a greater affinity for the bilayer than EGCg [80], and interactions division, cell wall turnover and cell separation appears to be related
between ECg and the hydrophobic core of the membrane can be to changes in the secretion of autolysins from the cell; ECg-grown
increased by structural modification of the natural product [81]. cells retain autolysins within the thickened cell wall, probably
Interactions between lipid bilayers and galloyl catechins such as in a predominantly inactive form, with greatly reduced amounts
ECg are facilitated by an exposed hydrophobic domain in the region released into the growth medium [85]. Activity and retention of
of the ester bond and C-ring, optimised by virtue of epi (cis) stereo- these enzymes, key players in cell separation and peptidoglycan
chemistry (Fig. 1). turnover, are strongly influenced by the charge characteristics of
Intercalation of these polyphenols into the cytoplasmic mem- the cell wall [89]. Growth in the presence of ECg decreases the
brane of S. aureus, including multidrug-resistant clinical isolates net positive charge of the cell surface (Fig. 2), in part by reducing
(MRSA), almost certainly accounts for the distinctive pheno- the degree of d-alanylation of wall teichoic acids associated with
type induced by ECg and, to a lesser extent, EGCg. Moderate peptidoglycan [90]. An increased negative surface charge leads to
(3–12.5 mg/L) concentrations of ECg completely abrogated ␤- electrostatic repulsion from negatively charged surfaces [89] and in
lactam resistance in all MRSA from a large collection of clinical all likelihood accounts for the biofilm-inhibiting properties of ECg.

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Fig. 2. Scanning electron micrographs of meticillin-resistant Staphylococcus aureus (MRSA) isolates (A,B) BB568 and (C,D) EMRSA-16 grown in non-supplemented medium
(A,C) and medium containing 12.5 mg/L (−)-epicatechin gallate (ECg) (B,D), then exposed to cationised ferritin. ECg-grown cells bind large quantities of the positively charged
probe.
These changes explain some aspects of the complex ECg-
induced MRSA phenotype, but the profound changes in ␤-lactam
susceptibility engendered by ECg are likely to be due to direct
interference with the antibiotic resistance machinery of the cell.
Similarly, loss of halotolerance and a markedly reduced capacity
to export proteins implies fundamental perturbation of key cel-
lular processes involving changes to the biophysical properties of
the cytoplasmic membrane. Staphylococcus aureus elaborates an
unusual membrane [91,92] comprising three major phospholipids:
negatively charged phosphatidylglycerol (PG) and cardiolipin in
addition to positively charged lysyl-PG (LPG), with evidence for
asymmetric distribution across the outer and inner cytoplasmic
membrane leaflets [92]. Exposure to ECg alters the expression of
genes encoding membrane-embedded proteins but also upregu-
lates genes belonging to the cell wall stress stimulon [52]. These
genes are induced following treatment with cell-wall-active antibi-
otics and cytoplasmic membrane depolarising agents such as
daptomycin [93,94], providing evidence that the bacteria respond
to ECg by taking steps to preserve and/or repair a compromised cell
wall and membrane. Indeed, the cell wall may represent an addi-
tional site of ECg binding, as a proportion (38%) of [3 H]ECg can be
recovered from the cell wall fraction, in addition to the membrane
(58%), after incubation with EMRSA-16 cells [52].
ECg rapidly enters the cytoplasmic membrane, causing an
immediate reduction in bilayer fluidity, but growing cells respond
by altering the composition of the membrane, incorporating a
greater complement of branched chain fatty acids to re-establish a
fluid phospholipid palisade. Membrane asymmetry is not compro-
mised by ECg (H. Rosado and P.W. Taylor, unpublished observation).
ECg intercalation appears to compromise membrane-anchored
MprF, as it engenders a large reduction in LPG content with a con-
comitant increase in PG [52]. This enzyme is responsible for the
attachment of lysine to PG and translocation of LPG to the outer
Please cite this article in press as: Taylor PW. Alternative natural sources for a new generation of antibacterial agents. Int J Antimicrob Agents
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5
leaflet of the membrane, where its positive-charge profile provides
protection against cationic antibacterial peptides and daptomycin
[95]. Thus, ECg may elicit reductions in virulence over and above
those due to large reductions in secretion of enzymes and toxins.
The elaboration of a more mobile and more negatively charged
bilayer radically alters the environment in which proteins involved
in cell wall biogenesis function and may be a key factor in the
perturbation of orderly wall assembly.
In dividing cells, PBP2-mediated transglycosylation and
transpeptidation reactions occur predominantly at the division
septum [96]. The PBP2 substrate within extending peptidoglycan
chains is restricted to the septum and PBP2 molecules are recruited
to this site by a process requiring the presence of FtsZ, a protein that
anchors the cell wall synthetic machinery to the site of cell division
[97]. ␤-Lactam antibiotics bind covalently to the transpeptidase
active site of PBP2 and abrogate catalytic activity, preventing for-
mation of pentaglycine cross-bridges and, in ␤-lactam-susceptible
strains, delocalising PBP2 from the septum [97]. As PBP2 and
PBP2a function co-operatively in the presence of oxacillin to
ensure orderly peptidoglycan synthesis at the septum, there is no
delocalisation of PBP2 in MRSA strains [97]. FtsZ remains at the
septum after exposure of MRSA to ECg, but ECg intercalation into
the bilayer delocalises PBP2 [52], functionally decoupling PBP2
and PBP2a (S. Paulin and P.W. Taylor, unpublished observation).
Thus, PBP2a is no longer able to compensate for the loss of PBP2
transpeptidase activity in the presence of oxacillin and this process
is highly likely to account for the reversible sensitisation of MRSA
strains by ECg. Decoupling of antibiotic resistance machineries by
membrane intercalators represents a novel and intriguing mecha-
nism for phenotypic modulation of drug resistance and raises the
possibility that combinations of ␤-lactam and intercalating agents
could be employed to restore the efficacy of second-generation
penicillins and cephalosporins whose utility has been severely
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6 P.W. Taylor / International Journal of Antimicrobial Agents xxx (2013) xxx–xxx
eroded by the emergence of resistance. With such a large number
of membrane-interactive plant metabolites available, it is likely
that other benign compounds will possess similar potentially
beneficial properties.
6. Conclusions
The re-emergence of evaluation of natural products as a source
of new drugs is particularly relevant to antibacterial drug discovery.
Historically, almost all antibiotics derived from secondary metabo-
lites are produced by soil bacteria and fungi, but it is unlikely
that such an approach will be precisely resurrected. However,
other environmental niches occupied by complex communities
of micro-organisms are only just beginning to be explored and,
in combination with genomics and advanced genetic engineering
[98], may provide new and exciting leads for the next generation
of antimicrobial agents. Too much effort has been expended on
what appears to be a fruitless search for plant-derived compounds
with properties that would permit development as conventional
antibiotics; resources would be used more profitably away from the
‘grind and find’ of traditional ‘herbal remedy’ approaches towards
the embrace of emerging molecular and chemical technologies.
Some plant-derived products do, however, have the potentially
valuable property of modifying complex bacterial phenotypes that
could benefit the new paradigms of antivirulence and resistance
modification that are currently in their infancy [83,99,100].
Funding: No funding sources.
Competing interests: None declared.
Ethical approval: Not required.
References
[1] Newman DJ, Cragg GM. Natural products as sources of new drugs over the
last 25 years. J Nat Prod 2007;70:461–77.
[2] Hopwood DA. Streptomyces in nature and medicine: the antibiotic makers.
New York, NY: Oxford University Press; 2007.
[3] Watve MG, Tickoo R, Jog MM, Bhole BD. How many antibiotics are produced
by the genus Streptomyces? Arch Microbiol 2001;176:386–90.
[4] Sams-Dodd F. Target-based drug discovery: is something wrong? Drug Discov
Today 2005;10:139–47.
[5] Payne DJ, Gwynn MN, Holmes DJ, Pompliano DL. Drugs for bad bugs: con-
fronting the challenges of antibacterial drug discovery. Nat Rev Drug Discov
2007;6:29–40.
[6] Sheridan C. Antibiotics au naturel. Nat Biotechnol 2006;24:1494–6.
[7] Zhu F, Ma XH, Qin C, Tao L, Liu X, Shi Z, et al. Drug discovery prospect from
untapped species: indications from approved natural product drugs. PLoS One
2012;7:e39782.
[8] Brötz-Oesterhelt H, Beyer D, Kroll H-P, Endermann R, Ladel C, Schroeder W,
et al. Dysregulation of bacterial proteolytic machinery by a new class of antibi-
otics. Nat Med 2005;11:1082–7.
[9] Wang J, Soisson SM, Young K, Shoop W, Kodali S, Galgoci A, et al. Platen-
simycin is a selective FabF inhibitor with potent antibiotic properties. Nature
2006;441:358–61.
[10] Fishbach MA, Walsh CT. Antibiotics for emerging pathogens. Science
2009;325:1089–93.
[11] Lewis K, Ausubel FM. Prospects for plant-derived antibacterials. Nat Bio-
technol 2006;24:1504–7.
[12] Dossey AT. Insects and their chemical weaponry: new potential for drug dis-
covery. Nat Prod Rep 2010;27:1737–57.
[13] Hancock REW, Sahl H-G. Antimicrobial and host-defense peptides as new
anti-infective therapeutic strategies. Nat Biotechnol 2006;24:1551–7.
[14] Leszczyńska K, Namiot D, Byfield FJ, Cruz K, Żendzian-Piotrowska M, Fein
DE, et al. Antibacterial activity of the human host defence peptide LL-37 and
selected synthetic cationic lipids against bacteria associated with oral and
upper respiratory tract infections. J Antimicrob Chemother 2013;68:610–8.
[15] Yim G, Wang HH, Davies J. Antibiotics as signalling molecules. Phil Trans R
Soc Lond B Biol Sci 2007;362:1195–200.
[16] Schloss PD, Handelsman J. Toward a census of bacteria in soil. PLoS Comput
Biol 2006;7:e92.
[17] Junge K, Eiken H, Deming JW. Bacterial activity at −2 to −20 ◦ C in Arctic
wintertime sea ice. Appl Environ Microbiol 2004;70:550–7.
[18] Price PB, Bay RC. Marine bacteria in deep Arctic and Antarctic ice cores: a
proxy for evolution in oceans over 300 million generations. Biogeosci Disc
2012;9:6535–77.
Please cite this article in press as: Taylor PW. Alternative natural sources for a new generation of antibacterial agents. Int J Antimicrob Agents
(2013), http://dx.doi.org/10.1016/j.ijantimicag.2013.05.004
[19] Beatty JT, Overmann J, Lince MT, Manske AK, Lang AS, Blankenship RE, et al. An
obligately photosynthetic bacterial anaerobe from a deep-sea hydrothermal
vent. Proc Natl Acad Sci U S A 2005;102:9306–10.
[20] Petersen JM, Zielinski FU, Pape T, Seifert R, Moraru C, Amann R, et al.
Hydrogen is an energy source for hydrothermal vent symbioses. Nature
2011;476:176–80.
[21] Forschner-Dancause S, LaPlante K, Smith DC, Rowley DC. Going deep for drug
discovery: an ocean to bedside approach to explore sub-seafloor microbes for
the next generation of antibiotics. Med Health R I 2012;95:292–3.
[22] Barton H. Introduction to cave microbiology: a review for the non-specialist.
J Caves Karst Stud 2006;68:43–54.
[23] Bastian F, Jurado V, Nováková A, Alabouvette C, Saiz-Jimenez C. The microbi-
ology of Lascaux cave. Microbiology 2010;156:644–52.
[24] Zinger L, Amaral-Zettler LA, Fuhrman JA, Horner-Devine MC, Huse SM, Welch
DBM, et al. Global patterns of bacterial ␤-diversity in seafloor and seawater
ecosystems. PLoS One 2011;6:e24570.
[25] Curtis TP, Sloan WT, Scannell JW. Estimating prokaryotic diversity and its
limits. Proc Natl Acad Sci U S A 2002;99:10494–9.
[26] Mayer AMS, Glaser KB, Cuevas C, Jacobs RS, Kem W, Little RD, et al. The odyssey
of marine pharmaceuticals: a current pipeline perspective. Trends Pharmacol
Sci 2010;31:255–65.
[27] Imhoff JF, Labes A, Wiese J. Bio-mining the microbial treasures of the ocean:
new natural products. Biotechnol Adv 2011;29:468–82.
[28] Kier G, Kreft H, Lee TM, Jetz W, Ibisch PL, Nowicki C, et al. A global assessment
of endemism and species richness across island and mainland regions. Proc
Natl Acad Sci U S A 2009;106:9322–7.
[29] Dixon RA. Natural products and plant disease resistance. Nature
2001;401:843–7.
[30] Shaw D. Risks or remedies? Safety aspects of herbal remedies in the UK. J R
Soc Med 1998;91:294–6.
[31] Paddon CJ, Westfall PJ, Pitera DJ, Benjamin K, Fisher K, McPhee D, et al.
High-level semi-synthetic production of the potent antimalarial artemisinin.
Nature 2013;496:528–32.
[32] Frankel OH, Brown AHD, Burdon JJ. The conservation of plant biodiversity.
Cambridge, UK: Cambridge University Press; 1995.
[33] Bode M, Payyappallimana U. Evidence based traditional medicine: for whom
and to what end? eJ Ind Med 2013;6:1–20.
[34] Mathez-Stiefel S, Vandenbroek I, Rist S. Can Andean medicine coexist with
biomedical healthcare? A comparison of two rural communities in Peru and
Bolivia. J Ethnobiol Ethnomed 2012;8:26.
[35] Tanaka MM, Kendal JR, Laland KN. From traditional medicine to witchcraft:
why medical treatments are not always efficacious. PLoS One 2009;4:e5192.
[36] World Health Organization. WHO traditional medicine strategy 2002–2005.
Geneva, Switzerland: WHO; 2002. WHO/EDM/TRM/2002.1.
[37] Hsieh SC, Lai JN, Chen PC, Chen HJ, Wang JD. Development of active safety
surveillance system for traditional Chinese medicine: an empirical study in
treating climacteric women. Pharmacoepidemiol Drug Saf 2006;15:889–99.
[38] Birks J, Grimley Evans J. Ginkgo biloba for cognitive impairment and dementia.
Cochrane Database Syst Rev 2009:CD003120.
[39] Coxeter PD, McLachlan AJ, Duke CC, Roufogalis BD. Herb–drug interactions:
an evidence-based approach. Curr Med Chem 2004;11:1513–25.
[40] Hsieh SC, Lai JN, Chen PC, Chen CC, Chen HJ, Wang JD. Is Duhuo Jisheng Tang
containing Xixin safe? A four-week safety study. Chin Med 2010;5:6.
[41] Timmer A, Günther J, Rücker G, Motschall E, Antes G, Kern WV. Pelargonium
sidoides extract for acute respiratory tract infections. Cochrane Database Syst
Rev 2008:CD006323.
[42] Naz R, Bano A. Antimicrobial potential of Ricinus communis leaf extracts in
different solvents against pathogenic bacterial and fungal strains. Asian Pac J
Trop Biomed 2012;2:944–7.
[43] Mohamed AA, Ali SI, El-Baz FK. Antioxidant and antibacterial activities
of crude extracts and essential oils of Syzygium cumini leaves. PLoS One
2013;8:e60269.
[44] Rubio-Moraga A, Argandoña J, Mota B, Pérez J, Verde A, Fajardo J,
et al. Screening for polyphenols, antioxidant and antimicrobial activ-
ities of extracts from eleven Helianthemum taxa (Cistaceae) used
in folk medicine in south-eastern Spain. J Ethnopharmacol 2013.
http://dx.doi.org/10.1016/j.jep.2013.04.028 [Epub ahead of print].
[45] Cowan MC. Plant products as antimicrobial agents. Clin Microbiol Rev
1999;12:564–82.
[46] Gibbons S. Anti-staphylococcal plant natural products. Nat Prod Rep
2004;21:263–77.
[47] Saleem M, Nazir M, Ali MS, Hussain H, Lee YS, Riaz N, et al. Antimicrobial
natural products: an update on future antibiotic drug candidates. Nat Prod
Rep 2010;27:238–54.
[48] O’Donnell G, Gibbons S. Antibacterial activity of two canthin-6-one alkaloids
from Allium neapolitanum. Phytother Res 2007;21:653–7.
[49] Maxwell A. The interaction between coumarin drugs and DNA gyrase. Mol
Microbiol 1993;9:681–6.
[50] Cushnie TPM, Lamb AJ. Antimicrobial activity of flavonoids. Int J Antimicrob
Agents 2005;26:343–56.
[51] Taylor PW, Hamilton-Miller JMT, Stapleton PD. Antimicrobial properties of
green tea catechins. Food Sci Technol Bull 2005;2:71–81.
[52] Bernal P, Lemaire S, Pinho MG, Mobashery S, Hinds J, Taylor PW. Insertion of
epicatechin gallate into the cytoplasmic membrane of methicillin-resistant
Staphylococcus aureus disrupts penicillin-binding protein (PBP) 2a-mediated
␤-lactam resistance by delocalizing PBP2. J Biol Chem 2010;285:24055–65.
 ARTICLE IN PRESS
G Model
ANTAGE-4105; No. of Pages 7
P.W. Taylor / International Journal of Antimicrobial Agents xxx (2013) xxx–xxx
[53] Moujir L, Gutiérrez-Navarro AM, González AG, Ravelo AG, Luis JG. The rela-
tionship between structure and antimicrobial activity in quinones from the
Celastraceae. Biochem Syst Ecol 1990;18:25–8.
[54] Ignacimuthu S, Pavunraj M, Duraipandiyan V, Raja N, Muthu C. Antibacterial
activity of a novel quinone from the leaves of Pergularia daemia (Forsk.), a
traditional medicinal plant. Asian J Trad Med 2009;4:36–40.
[55] Pandurangan K, Murnaghan KD, Walsh A, Müller-Bunz H, Paradisi F, Mor-
gan CG. Design, synthesis and structure of novel para-quinones and their
antibacterial activity. Chem Biol Drug Des 2011;78:787–99.
[56] Bolton JL, Trush MA, Penning TM, Dryhurst G, Monks TJ. Role of quinones in
toxicology. Chem Res Toxicol 2000;13:135–60.
[57] Gershenzon J, Dudareva N. The function of terpene natural products in the
natural world. Nat Chem Biol 2007;3:408–14.
[58] Alves DS, Pérez-Fons L, Estepa A, Micol V. Membrane-related effects under-
lying the biological activity of the anthraquinones emodin and barbaloin.
Biochem Pharmacol 2004;68:549–61.
[59] Caturla N, Pérez-Fons L, Estepa A, Micol V. Differential effects of oleuropein,
a biophenol from Olea europaea, on anionic and zwitterionic phospholipid
model membranes. Chem Phys Lipids 2005;137:2–17.
[60] Laporta O, Funes L, Garzón MT, Villalaín J, Micol V. Role of membranes on the
antibacterial and anti-inflammatory activities of the bioactive compounds
from Hypoxis rooperi corm extract. Arch Biochem Biophys 2007;467:119–31.
[61] Stapleton PD, Shah S, Anderson JC, Hara Y, Hamilton-Miller JMT, Taylor PW.
Modulation of ␤-lactam resistance in Staphylococcus aureus by catechins and
gallates. Int J Antimicrob Agents 2004;23:462–7.
[62] Caturla N, Vera-Samper E, Villalain J, Reyes Mateo C, Micol V. The relationship
between the antioxidant and antibacterial properties of galloylated catechins
and the structure of phospholipid model membranes. Free Radic Biol Med
2003;34:648–62.
[63] Guest DI, Brown JF. Plant defences against pathogens. In: Brown JF, Ogle JH,
editors. Plant pathogens and plant diseases. Armidale, NSW: Rockvale Publi-
cations; 1997. p. 263–86.
[64] Freeman BC, Beattie GA. An overview of plant defenses
against pathogens and herbivores. Plant Health Instruct 2008.
http://dx.doi.org/10.1094/PHI-I-2008-0226-01.
[65] Regnault-Roger C. The potential of botanical essential oils for insect pest
control. Integr Pest Manag Rev 1997;2:25–34.
[66] Compant S, Duffy B, Nowak J, Clément C, Barka EA. Use of plant growth-
promoting bacteria for biocontrol of plant diseases: principles, mechanisms
of action, and future prospects. Appl Environ Microbiol 2005;71:4951–9.
[67] Hammerschmidt R. Phytoalexins: what have we learned after 60 years? Annu
Rev Phytopathol 1999;37:285–306.
[68] Kagan IA, Hammerschmidt R. Arabidopsis ecotype variability in camalexin
production and reaction to infection by Alternaria brassicicola. J Chem Ecol
2002;28:2121–40.
[69] Noutoshi Y, Okazaki M, Kida T, Nishina Y, Morishita Y, Ogawa T, et al. Novel
plant immune-priming compounds identified via high-throughput chemical
screening target salicylic acid glucosyltransferases in Arabidopsis. Plant Cell
2012;24:3795–804.
[70] Noutoshi Y, Okazaki M, Shirasu K. Isolation and characterization of the plant
immune-priming compounds Imprimatin B3 and -B4, potentiators of disease
resistance in Arabidopsis thaliana. Plant Signal Behav 2012;7:1526–8.
[71] Aranda FJ, Villalaín J. The interaction of abietic acid with phospholipid mem-
branes. Biochim Biophys Acta 1997;1327:171–80.
[72] Micol V, Mateo CR, Shapiro S, Aranda FJ, Villalaín J. Effects of (+)-totarol, a
diterpenoid antibacterial agent, on phospholipid model membranes. Biochim
Biophys Acta 2001;1511:281–90.
[73] Stermitz FR, Lorenz P, Tawara JN, Zenewicz LA, Lewis K. Synergy in
a medicinal plant: antimicrobial action of berberine potentiated by 5 -
methoxyhydnocarpin, a multidrug pump inhibitor. Proc Natl Acad Sci U S
A 2000;97:1433–7.
[74] Lomovskaya O, Zgurskaya HI, Totrov M, Watkins WJ. Waltzing transporters
and ‘the dance macabre’ between humans and bacteria. Nat Rev Drug Discov
2007;6:56–65.
[75] Lomovskaya O, Bostian KA. Practical applications and feasibility of efflux
pump inhibitors in the clinic—A vision for applied use. Biochem Pharmacol
2006;71:910–8.
[76] Garvey MI, Piddock LJV. The efflux pump inhibitor reserpine selects
multidrug-resistant Streptococcus pneumoniae strains that overexpress
the ABC transporters PatA and PatB. Antimicrob Agents Chemother
2008;52:1677–85.
[77] Kajiya K, Kumazawa S, Nakayama T. Steric effects on the interaction of tea
catechins with lipid bilayers. Biosci Biotechnol Biochem 2001;65:2638–43.
Please cite this article in press as: Taylor PW. Alternative natural sources for a new generation of antibacterial agents. Int J Antimicrob Agents
(2013), http://dx.doi.org/10.1016/j.ijantimicag.2013.05.004
7
[78] Kajiya K, Kumazawa S, Nakayama T. Effects of external factors on the
interaction of tea catechins with lipid bilayers. Biosci Biotechnol Biochem
2002;66:2330–5.
[79] Stapleton PD, Shah S, Hara Y, Taylor PW. Potentiation of catechin gallate-
mediated sensitization of Staphylococcus aureus to oxacillin by nongalloylated
catechins. Antimicrob Agents Chemother 2006;50:752–5.
[80] Hashimoto T, Kumazawa S, Nanjo F, Hara Y, Nakayama T. Interaction of tea
catechins with lipid bilayers investigated with liposome systems. Biosci Bio-
technol Biochem 1999;63:2252–5.
[81] Anderson JC, McCarthy R, Paulin S, Taylor PW. Anti-staphylococcal activity
and antibiotic resistance attenuating capacity of structural analogues of (−)-
epicatechin gallate. Bioorg Med Chem Lett 2011;21:6996–7000.
[82] Shah S, Stapleton PD, Taylor PW. The polyphenol (−)-epicatechin gallate dis-
rupts the secretion of virulence-related proteins by Staphylococcus aureus.
Lett Appl Microbiol 2008;46:181–5.
[83] Taylor PW, Bernal P, Zelmer A. Modification of the bacterial phenotype as
an approach to counter the emergence of multidrug-resistant pathogens.
In: Bonilla AR, Muniz KP, editors. Antibiotic resistance: causes and risk fac-
tors, mechanisms and alternatives. Hauppauge, NY: Nova Science Publishers;
2009. p. 43–78.
[84] Blanco AR, Sudano-Roccaro A, Spoto GC, Nostro A, Rusciano D. Epigallocat-
echin gallate inhibits biofilm formation by ocular staphylococcal isolates.
Antimicrob Agents Chemother 2005;49:4339–43.
[85] Stapleton PD, Shah S, Ehlert K, Hara Y, Taylor PW. The ␤-lactam-resistance
modifier (−)-epicatechin gallate alters the architecture of the cell wall of
Staphylococcus aureus. Microbiology 2007;153:2093–103.
[86] Stapleton PD, Gettert J, Taylor PW. Epicatechin gallate, a component of green
tea, reduces halotolerance in Staphylococcus aureus. Int J Food Microbiol
2006;111:276–9.
[87] Fuda C, Hesek D, Lee M, Morio K, Nowak T, Mobashery S. Activation for cataly-
sis of penicillin-binding protein 2a from methicillin-resistant Staphylococcus
aureus by bacterial cell wall. J Am Chem Soc 2005;127:2056–7.
[88] Pinho MG, Filipe SR, de Lencastre H, Tomasz A. Complementation of the
essential peptidoglycan transpeptidase function of penicillin-binding protein
2 (PBP2) by the drug resistance protein PBP2A in Staphylococcus aureus. J
Bacteriol 2001;183:6525–31.
[89] Neuhaus FC, Baddiley J. A continuum of anionic charge: structures and func-
tions of d-alanyl-teichoic acids in Gram-positive bacteria. Microbiol Mol Biol
Rev 2003;67:686–723.
[90] Bernal P, Zloh M, Taylor PW. Disruption of d-alanyl esterification
of Staphylococcus aureus cell wall teichoic acid by the ␤-lactam-
resistance modifier (−)-epicatechin gallate. J Antimicrob Chemother 2009;63:
1156–62.
[91] Haest CW, de Gier J, op den Kamp JA, Bartels P, van Deenen LL. Changes in
permeability of Staphylococcus aureus and derived liposomes with varying
lipid composition. Biochim Biophys Acta 1972;255:720–33.
[92] Mukhopadhyay K, Whitmire W, Xiong YQ, Molden J, Jones T, Peschel A, et al.
In vitro susceptibility of Staphylococcus aureus to thrombin-induced platelet
microbicidal protein-1 (tPMP-1) is influenced by cell membrane phospholipid
composition and asymmetry. Microbiology 2007;153:1187–97.
[93] McAleese F, Wu SW, Sieradski K, Dunman P, Murphy E, Projan S, et al. Overex-
pression of genes of the cell wall stimulon in clinical isolates of Staphylococcus
aureus exhibiting vancomycin-intermediate S. aureus-type resistance to van-
comycin. J Bacteriol 2006;188:1120–33.
[94] Muthaiyan A, Silverman JA, Jayaswal RK, Wilkinson BJ. Transcriptional pro-
filing reveals that daptomycin induces the Staphylococcus aureus cell wall
stress stimulon and genes responsive to membrane depolarization. Antimi-
crob Agents Chemother 2008;52:980–90.
[95] Ernst CM, Staubitz P, Mishra NN, Yang SJ, Hornig G, Kalbacher H, et al. The
bacterial defensin resistance protein MprF consists of separable domains
for lipid lysinylation and antimicrobial peptide repulsion. PLoS Pathog
2009;5:e1000660.
[96] Pinho MG, Errington J. Dispersed mode of Staphylococcus aureus cell
wall synthesis in the absence of the division machinery. Mol Microbiol
2003;50:871–81.
[97] Pinho MG, Errington J. Recruitment of penicillin-binding protein PBP2 to the
division site of Staphylococcus aureus is dependent on its transpeptidation
substrates. Mol Microbiol 2005;55:799–807.
[98] Grushkin D. Natural products emergent. Nat Med 2013;19:390–2.
[99] Projan SJ, Youngman PJ. Antimicrobials: new solutions badly needed. Curr
Opin Microbiol 2002;5:463–5.
[100] Rasko DA, Sperandio V. Anti-virulence strategies to combat bacteria-
mediated disease. Nat Rev Drug Discov 2010;9:117–28.