Microbiological Research 165 (2010) 437—449

www.elsevier.de/micres

Recent developments and future prospects of

antimicrobial metabolites produced by endophytes
Hongsheng Yu, Lei Zhang, Lin Li, Chengjian Zheng, Lei Guo, Wenchao Li,
Peixin Sun, Luping Qin

Department of Pharmacognosy, School of Pharmacy, Second Military Medical University, 325 Guohe Road, Shanghai
200433, PR China

Received 1 July 2009; received in revised form 19 November 2009; accepted 21 November 2009

KEYWORDS Summary
Endophyte;
Endophytes, found ubiquitous in all plant species in the world, contribute to their
Natural product;
host plants by producing plenty of substances that provide protection and ultimately
Antimicrobial com-
survival value to the plant. Many researches have proven that endophyte is a new
pound
and potential source of novel natural products for exploitation in modern medicine,
agriculture and industry. So far, a great number of novel natural products possessing
antimicrobial activities have been isolated from endophytes. It is believed that
screening for antimicrobial compounds from endophytes is a promising way to
overcome the increasing threat of drug resistant strains of human and plant
pathogen. Antimicrobial metabolites isolated from endophytes belong to diverse
structural classes, including: alkaloids, peptides, steroids, terpenoids, phenols,
quinones, and flavonoids. In this review, many well-studied areas are presented and
examples will be given to discuss the structure of compounds isolated from
endophytes extracts with antimicrobial activities and show the wide variety of
approaches taken within the field. These achievements would provide the
opportunity to utilize endophytes as a new source for production of antibiotics.
& 2009 Elsevier GmbH. All rights reserved.

Contents

1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 438
2. Methods of searching for antimicrobial compounds from endophytes . . . . . . . . . . . . . . . . . . . . . . . . . 438
2.1. Rationale for plant selection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 438

Corresponding author. Tel./fax: þ86 21 81871300.

E-mail address: qinsmmu@126.com (L. Qin).

0944-5013/$ - see front matter & 2009 Elsevier GmbH. All rights reserved.
doi:10.1016/j.micres.2009.11.009
438 H. Yu et al.

2.2. Screening antimicrobial natural products. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 439

3. Biological survey of endophytic natural products . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 442
3.1. Alkaloids . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 442
3.2. Peptides . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 442
3.3. Steroids. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 442
3.4. Terpenoids . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 442
3.5. Phenols . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 444
3.6. Quinones . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 445
3.7. Aliphatic compounds . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 446
3.8. Flavonoids . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 446
3.9. Others. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 447
4. Conclusion and future targets . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 447
Acknowledgements. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 447
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 448

1. Introduction either microbial metabolites or their semi-synthetic

Food production needs to be increased to meet the
rapidly expanding world population. Unfortunately,
severe cereal loss is still inevitable owing to plant
diseases (Aliye et al. 2008), particularly those caused
by phytopathogen. Human often face problems of
the tremendous increase in the incidence of fungal
or bacterial infections in the world’s population.
Chemical synthetic drugs with many side effects are
being used to cope with these medical problems
(Strobel and Daisy, 2003). Both human pathogens and
phytopathogens are prone to develop ‘‘drug’’ resis-
tances to decrease substantially the effectiveness of
old antibiotics. Accordingly, there is an urgent need
to work towards the invention of safer antifungal
agents which are expected to be renewable, non-
petrochemical, naturally eco-friendly and easily
obtainable (Liu et al., 2001).
Endophyte, by definition, is one which resides in
the tissues beneath the epidermal cell layers and
causes no apparent harm to the host (Stone et al.
2000). They form inconspicuous infections within
tissues of healthy plants for all or nearly all their life
cycle (Limsuwan et al. 2009). Studies have shown
that, nearly 300,000 plant species that exist on the
earth, each individual plant is the host to one or
more endophytes, the population of a given en-
dophytic species varies from several to a few
hundreds strains (Strobel and Daisy 2003; Huang
et al. 2007). There is a complex relationship
between endophytes and their host plants. It is
believed that endophytes indirectly benefit plant
growth by producing special substances mainly
secondary metabolites to prevent the growth or
activity of plant pathogens. In support of this idea,
metabolites of endophytes have been reported to
inhibit a number of microorganisms (Fisher et al.
1984; Gurney and Mantle 1993). Many important
antifungal and antibacterial chemotherapeutics are
derivatives. It is estimated that there might be as
many as one million different endophyte species,
however only a handful of them have been described
(Petrini 1991), which means investigating the
metabolites of endophytes can increase the chance
of finding novel antimicrobial natural products.
Heretofore, studies reported hundreds of bioactive
natural products, a larger proportion of which
showed antimicrobial activities, have been isolated
from endophytes. The aim of the review is to take a
comprehensive look at the most recent research on
antimicrobial metabolites produced by endophytes.
2. Methods of searching for
antimicrobial compounds from
endophytes
2.1. Rationale for plant selection
It is important to understand the methods and
rationale used to provide the best opportunities to
isolate endophytes prone to produce novel anti-
microbial metabolites from so great number of
plant species. Several reasonable plant selection
strategies should be followed:
(1) Plants growing in areas of great biodiversity
also have the prospect of housing endophytes
with great biodiversity (Strobel et al. 2004).
Thus, tropical rainforest possessing the greatest
biodiversity on the earth is an endophyte
resources storehouse.
(2) Plants growing in special habitats, especially
those in deteriorated ecological environment,
and possessing special capabilities for survival
should also be selected for study. People may
learn that the power of plants living in such
Recent developments and future prospects of antimicrobial metabolites produced by endophytes
environment may get from endophytes reside in
the tissues of the plants. One example is the
special ecosystem mangrove forest growing on
the tidal shallows of sea border. Kumaresan and
Suryanarayanan (2001) studied mangrove for-
ests, and 12 species endophyte were isolated.
(3) Plants surrounded by pathogens infected plants
showing no symptoms are more likely to lodge
endophytes possessing antimicrobial natural
products than other plants. Tuntiwachwuttikul
et al. (2008) reported an endophyte showed
antimicrobial activity against plant pathogen
Colletotrichum musae.
(4) Plants that have been exploited for human use
as traditional medicines in some place should
be considered for study. Ji et al. (2005) isolated
an endophytic fungus possessing antimicrobial
activity from a traditional Chinese medicine
Celastrus angulatus.
(5) Plants occupied a certain ancient land mass,
are also more likely to lodge endophytes with
active natural products than other plants
(Strobel and Daisy 2003). Chaetoglobosin B
was isolated from chaetomium globosum in leaf
of Maytenus hookeri, which only distribute in
areas of Yunnan, China, showed antituberculo-
sis activity (Ni et al. 2008).
A comparison of the endophytic hosts (Figure 1)
shows that medical plants, crops, and plants in
special environment were frequently studied for
screening antimicrobial enophytes up to date.
2.2. Screening antimicrobial natural
products
Crude extracts from the fermentation broth of
each endophyte need firstly to be tested through
other plants 18%
plants in special
environment 18%
crops 29%
Figure 1. Proportion of biologically active isolates from different sources tested for antimicrobial activities in recent
researches. More interest has been devoted to screen endophytic fungi possessing antimicrobial activities from medical
plants and crops, resenting 35% and 29%, respectively.
439
some commonly used methods such as paper disk
diffusion method, agar dilution assay, disk diffusion
assay, and mycelial radial growth test (Zeng et al.
2005; Zhang et al. 2005; Gong et al. 2007; Hoffman
et al. 2008; Aly et al. 2008; Pongcharoen et al.
2008).
A plenty of researches have reported that
endophytes with no certain compound isolated
showed antimicrobial activities, and some of which
were significant. Five of sixty-three endophytic
fungi, isolated from the rhizomes of a traditional
medicinal herb Paris polyphylla var. yunnanensis,
showed strong antibacterial activities against four
Gram-negative bacteria (Escherichia coli, Xantho-
monas vesicatoria, Agrobacterium tumefaciens and
Pseudomonas lachrymans) and two Gram-positive
bacteria (Bacillus subtilis, Staphylococcus haemo-
lyticus) (Zhao et al. 2008). 353 endophytic fungi,
with no certain compound isolated, were separated
from medical plants (Cyrtomium, Ophiopogon).
The proportion of inhibiting the growth of Alter-
naria solani and Botrytis cinerea was 15.9% and
11.3%, respectively (Jiang et al. 2006). Phongpai-
chit et al. (2006) obtained the fermentation broths
of endphytic fungi from plants of Garcinia and
tested the antimicrobial activity through the agar
diffusion method against Staphylococcus aureus,
Candida albicans and Cryptococcus neoformans.
The results showed that 70 strains of the total 377
(18.6%) displayed antimicrobial activity against at
least one pathogenic microorganism. A mass of
endophyte researches are still in the screening
stage, and the active ingredients need to be
isolated.
Some endophytes strains showed antimicrobial
activities also have been found in our laboratory
from different plants (Scapania verrucosa, Panax
ginseng and Rehmannia glutinosa) (Guo et al. 2008;
Xu et al. 2009; You et al. 2009).
medical plants 35%
440 H. Yu et al.

Separation of active substances should be the
next step after the crude extracts of certain
endophytes have been proved to possess bioactiv-
ities against tested microbes.
Thin layer chromatography (TLC) and bio-auto-
graphy can be used to track antimicrobial ingre-
dients. The crude extract should be placed on the
TLC plate and then separated on the plate by
suitable liquid solvent. Tested microorganisms
(bacteria or fungal spores) are inoculated onto
the TLC plate that has separated components, and
then the microbial cultivation is carried out in the
moist environment for a few days. Antimicrobial
substances could be tracked through observation of
the inhibitory zone on the TLC plate. Ultimately,
the active components can be isolated through
commonly used methods (precipitation thin-layer
chromatography, liquid preparation chromatogra-
phy, and column chromatography, etc) (Han et al.
2008; Xu et al. 2008).
The efficacy of antimicrobial ingredient is in-
dicated by minimal inhibitory concentration (MIC)
(Tuntiwachwuttikul et al. 2008) or 50% inhibitory
concentrations (IC50) value (Macı́as-Rubalcava
et al., 2008). Tracked antimicrobial compounds
are examined with triphenyltetrazolium chloride
(TTC) or methyl thiazolyl tetrazolium (MTT) meth-
ods (Abe and Matsuki 2000). The general procedure
of antimicrobial compounds separation is showed in
Figure 2.
Examples of the antimicrobial activities of
various endophytes are showed in Table 1. As
shown in Table 1, we can conclude that
Pestalotiopsis sp., Streptomyces sp. and
Phomopsis sp. were the most frequently isolated
endophytes from host plants to possess
antimicrobial activities and the tested microbes
commonly used in previous researches including:
C. albicans, S. aureus, E. coli, A. sp., Fusarium sp.,
and Pyricularia oryzae, etc.

Plants
Rationale for plant
selection

Plants with high research value Plants with low research value

Rationale for isolation of

endophyte

Endophyte strains

Testing of antimicrobial
activity of crude extracts

Strains with unconspicuous Strains with significant

antimicrobial activity antimicrobial activity
Isolation of chemical
constituents

Chemical constituents
Testing of antimicrobial
activity

Antimicrobial Constituents Antimicrobial Constituents

Further research

Figure 2. Flow chart of antimicrobial compounds separation from endophytes.

Recent developments and future prospects of antimicrobial metabolites produced by endophytes 441

Table 1. Examples of the endophytic activities against microbes.

Types of Host plant Endophyte Test microbes Ref.

antimicrobial
compounds

Aliphatic Excoecaria agallocha Phomopsis sp. Candida albicans and Fusarium oxysporum Huang et al.
compounds (2008)
Ginkgo biloba Chaetomium Mucor miehei Qin et al.
globosum (2009)
Quercus variabilis Cladosporium sp Trichophyton rubrum, Candida albicans, Wang et al.
Aspergillus niger, Epidermophyton (2006)
floccosum, Microsporum canis

Alkaloids Garcinia dulcis Phomopsis sp. Mycobacterium tuberculosis Rukachaisirikul

et al. (2008)
Ginkgo biloba Chaetomium Mucor miehei Qin et al.
globosum (2009)
maize Acremonium Aspergillus flavus, Fusarium verticillioides Wicklow et al.
zeae (2005)

Flavonoids Juniperus cedre Nodulisporium Bacillus megaterium, Microbotryum Dai et al.

sp. violaceum, Septoria tritici, Chlorella (2006)
fusca
Peptides Acrostichum aureurm Penicillium sp. Staphylococcus aureus, Candida albicans Cui et al.
(2008)
Pinus sylvestris and Cryptosporiopsis yeasts Noble et al.
Fagus sylvatica sp., Pezicula sp. (1991)
Tripterigium wiflordii Cryptosporiopsis Candida albicans Strobel et al.
quercina (1999)
Tropical tree and vine Muscodor albus Candida albicans Strobel et al.
species in several of the (1999)
world’s rainforests
Phenols Cerbera manghas Penicillium sp. Staphylococcus aureus Han et al.
(2008)
Saurauia scaberrinae Phoma species Staphylococcus aureus Hoffman et al.
(2008)
Unidentified Pestalotiopsis Fusarium culmorum, Gibberella zeae, and Li et al. (2008)
adusta Verticillium aiboatrum
Quinones Callicarpa acuminata Edenia Phythophtora capsici, Phythophtora Macı́as-
gomezpompae parasitica, Fusarium oxysporum and Rubalcava
Alternaria solani et al. (2008)
Unidentified Pestalotiopsis Fusarium culmorum, Gibberella zeae, and Li et al. (2008)
adusta Verticillium aiboatrum
Urospermum picroides Ampelomyces sp. Staphylococcus aureus, Staphylococcus Aly et al.
epidermidis and Enterococcus faecalis (2008)

Steroids Artemisia annua Colletotrichum Phytophthora capisici, Rhizoctonia Lu et al. (2000)

sp. cerealis, Gaeumannomyces graminis var.
tritici, and Helminthosporium sati6um.
Juniperus cedre Nodulisporium Bacillus megaterium, Microbotryum Dai et al.
sp. violaceum, Septoria tritici, Chlorella (2006)
fusca

Terpenoids Cassia spectabilis Phomopis cassiae Cladosporium sphaerospermum and Silva et al.
Cladosporium cladosporioides (2006)
Daphnopsis americana Not identifide Staphylococcus aureus and Enterococcus Brady et al.
faecalis (2001)
Daphnopsis Americana Not mention Staphylococcus aureus and Enterococcus Brady et al.
faecalis (2000)
442
Table 1. (continued )
Types of Host plant Endophyte
antimicrobial
compounds
Taxus cuspidate Periconia sp.
3. Biological survey of endophytic
natural products
A mass of bioactive natural products isolated
from endophytes have been reported in recent few
years, among them antimicrobial active substances
including: alkaloids, peptides, steroids, terpenoids,
quinones, flavonoids, aliphatic compounds, and
phenols, etc (Figure 3).
3.1. Alkaloids
Alkaloids are quite common secondary metabolites
in endophytes, and some of them show antimicrobial
activities. Chaetoglobosins A (1) and C (2), were
characterized from the culture of an endophytic
C. globosum originating from the leaves of Ginkgo
biloba. In agar diffusion method, these two metabo-
lites were shown to be antibacterial against Mucor
miehei, further research show that both of them
may have harmful impact on mammals for showing
significant toxicity towards brine shrimp larvae (Qin
et al. 2009). Two newly reported antibiotics, pyrro-
cidines A (3) and B (4) were isolated from endophyte
Acremonium zeae in maize, displayed significant
antifungal activity against Aspergillus flavus and
Fusarium Verticillioides. Interactions between
A. zeae and other endophytes were considered and
the significance of these interactions relative to the
aflatoxin and fumonisin contamination in preharvest
maize was presented (Wicklow et al., 2005).
Phomoenamide (5) synthesized by an endophytic
fungi Phomopsis sp. in Garcinia dulcis exhibited the
MIC value of 6.25 mg/mL against Mycobacterium
tuberculosis. The antimicrobial activity of the meta-
bolite was significant (Rukachaisirikul et al. 2008).
A mass of alkaloids metabolite synthesized by
endophytes show significant antibacterial activities,
further researches are required.
3.2. Peptides
Many peptides produced by endophytes displayed
significant antimicrobial activities. Leuesnostatin A
(6) produced by Acremonium sp. in Taxus baccata,
H. Yu et al.
Test microbes Ref.
Bacillus subtilis, Staphylococcus aureus, Kim et al.
Klebsiella pneumoniae, and Salmonella (2004)
typhimurium
displayed antimicrobial activity against Pythium
ultimum (Strobel et al. 1997). Cryptocandin (7), a
novel peptide produced by endophytic Cryptospor-
iopsis quercina from Tripterigium wiflordii showed
antibacterial activity against C. albicans (Strobel
et al. 1999). A group of peptides, echinocandins (A,
B, C, D, H) (8, 9, 10, 11) isolated from endophytic
Cryptosporiopsis sp. and Pezicula sp. in Pinus
sylvestris and Fagus sylvatica showed to be anti-
microbial (Noble et al. 1991). Cyclo(Pro-Thr) (12)
and cyclo (Pro-Tyr) (13) were two new antibacterial
constituents produced by the fermentation broth of
endophytic fungus Penicillium sp. isolated from
mangrove plant Acrostichum aureurm (Cui et al.
2008). There are many reports on peptides in
earlier years, while rare about it in recent years.
Like the large number of antibacterial cationic
peptides secreted by insets, there is of great
application value to develop antibacterial drugs
from endophyte.
3.3. Steroids
Antimicrobial compounds ergosterol (14) and 5a,
8a-epidioxyergosterol (15) were isolated from the
culture extract of the endophytic fungus Nodulis-
porium sp. existed in Juniperus cedre on Gomera
Island (Dai et al. 2006). Four antimicrobial steroids
3b-hydroxy-ergosta-5-ene (16), 3-oxo-ergosta-4, 6,
8 (14), 22-tetraene (17) 3b, 5a-dihydroxy-6b-
acetoxy-ergosta-7, 22-diene (18), and 3b, 5a-
dihydroxy-6b-phenylacetyloxy-ergosta-7, 22-diene
(19), separated from Colletotrichum sp. in Artemi-
sia annua, showed fungistatic activities to the crop
pathogenic fungi Phytophthora capisici, Rhizocto-
nia cerealis, Gaeumannomyces graminis var. tritici,
and Helminthosporium sativum (Lu et al. 2000).
Most steroid compounds isolated from endo-
phytes only showed moderate antimicrobial activ-
ities, so it is difficult to find effective drugs or
pesticides from endophyte.
3.4. Terpenoids
Sesquiterpenes, diterpenoids and triterpenoids
are the major terpenoids isolated from endophytes.
Recent developments and future prospects of antimicrobial metabolites produced by endophytes 443

Some sesquiterpenes were proved to be antimicro- perieoniein B (23) were four novel diterpenoid
bial compounds up to present. Guanancastepene A antibiotics isolated from endophytes (Brady et al.
(20), guanacastepene (21) periconicin A (22), and 2000, 2001; Kim et al. 2004). Five cadinane

O O

H H

N N
O OH O
N O N O O
O O
1 Chaetoglobosins A 2 Chaetoglobosins C
H
H
O
O
H
H O O H
NH H O O
NH
OH
OH
3 Pyrrocidines A 4 Pyrrocidines B

O OH
H
N
N
H
OH O
5 Phomoenamide
OH

H3 CH 2 COCH 2 C(HO)HCH 2 C
O O OH OH
H O HO O
H
CO N
N N ON H2 N N
N H H H NH OH
H N H
NO O N
H HO O
O O
CO O N
O OH O
NH O NH OH
HO
NH N HN
N
H OH NH
O O
(CH 2 )14 CH 3
6 LeuesnostatinA 7 Cryptocandin
O OH
R2 R3 H
OH N
NH
NHCO H
O O O
HN 12 Cyclo(Pro-Thr)
HO
O
O
H
HH N OH N
HO O N
NH
H OH
O H
O
OH
R1

8 Enhinoandin A R 1 =R 2 =R 3 =OH 13 Cyclo (Pro-Tyr)

9 Enhinoandin B R 1 =H,R 2 =R 3 =OH
10 Enhinoandin C R 1 =R 2 =R 3 =H
11 Enhinoandin D R 1 =R 2 =OH, R3 =OMe

Figure 3. Antimicrobial alkaloids (1–5); peptides (6–13); steroids (14–19); terpenoids (20–28); phenols (29–35); quinones
(36–40); aliphatic compouds (41–45); flavonoids (46–48); and some other antimicrobial compouds (49–52) from
endophytes.
444 H. Yu et al.

O
O
HO HO
14 Ergosterol 15 5α,8α-epidioxyergosterol

HO O
16 3β-hydroxy-ergosta-5-ene 17 3-oxo-ergosta-4,6,8(14),22-tetraene

HO
OH HO
OCOCH 3 OH
OCOCH 2 C 6 H5

18 3β, 5α-dihydroxy-6β- 19 3β, 5α-dihydroxy -6β-

acetoxy-ergosta-7, 22-diene phenylacetyloxy -ergosta-7, 22-
diene
O OH
O
OH O OH OHC
O H O
O O O H
H

HO

20 Guanacastepene A 21 Guanacastepene 22 Periconicin A

OH CH 3 CH 3
OHC
HO OH HO OH
O H
H
H3 C H3 C
HO HO HO
CH 3 CH 3
23 Perieoniein B 24 25
CH 3 CH 3
CH 3
HO HO
HO

H3 C H3 C H3 C

HO HO HO
CH 3 CH 3 CH 3
OH
26 27 28

Figure 3. (Continued)

sesquiterpenes (24–28) derivatives were isolated
by bioassay-guided fractionation from Phomopis
cassiae, an endophytic fungus isolated from Cassia
spectabilis. They were evaluated against Clados-
porium sphaerospermum and C. cladosporioides,
revealing compound (28) as the most active
compound (Silva et al. 2006).
3.5. Phenols
Phenol and phenolic acids have often been
isolated from some endophytes cultures originating
from a variety of host plants. Two new antibiotics,
pestalachloride A (29) and B (30), have been isolated
from endophytic Pestalotiopsis adusta, which dis-
played significant antifungal activity against three
plant pathogens (Fusarium culmorum, Gibberella
zeae and Verticillium aiboatrum) (Li et al. 2008).
Hoffman et al. (2008) isolated a group of phenolic
acids (31–33) from culture broth of a Phoma sp.,
discovered as an endophyte on a Guinea plant
(Saurauia scaberrinae). The compounds exhibited
antibacterial activities against gram-positive bacter-
ia (S. aureus, P.ultimum, Sclerotinia sclerotiorum
and Rhizoctonium solani). Han et al. (2008) reported
that two new phenol antibiotics (34, 35) with
Recent developments and future prospects of antimicrobial metabolites produced by endophytes 445

OH OH
O
Cl
O O O
OH
NH
OH OH OH
O
Cl O
OH OH O
HO
Cl
Cl O

29 Pestalachloride A 30 Pestalachloride B 31

O O O O
OH O
H3 C
OH OH

O OCH O
HO 3 HO O

O NH2 O
32 Cercosporamide 33 Phomodione
OH

HO
OH
HO
OH

O OH
OH O OH O OH
OH
HO H3 CO

OCH 3 OH O O O
O OH O OH

36 3-O-methylalaternin 37 Altersolanol A
O
38
O OH O OH

O O O O O O
OH OCH 3

O O
39 40

Figure 3. (Continued)

activities against methicillin-resistant S. aureus,
produced by Penicillium sp. isolated as an endo-
phyte from mangrove plant Cerbera martghas.
Many novel phenol and phenolic acids, isolated
from endophytes cultures in recent studies, often
have showed pronounced antimicrobial activities.
With further studies on phenol and phenolic acids,
there will be a great potential to find newly
effective antibiotics.
3.6. Quinones
3-O-methylalaternin (36) and altersolanol A
(37) were synthesized by Ampelomyces sp., an
endophyte isolated from the medicinal plant
Urospermum picroides, displayed antimicrobial
activity against the gram-positive pathogens,
S. aureus, S.epidermidis and Enterococcus faecalis
at MIC value of 12.5 and 12.5–25 mg/mL, respec-
tively. The antibacterial activity of altersolanol A is
probably not due to its cytotoxic activity, but for
the compound acts as an electron acceptor in the
bacterial membrane and thus inhibits bacterial
growth (Haraguchi et al. 1992; Aly et al. 2008).
Macı́as-Rubalcava et al. (2008) firstly reported that
three novel spiroketals (38, 39, 40), isolated from a
newly discovered endophytic fungus Edenia gomez-
pompae, displayed significant growth inhibition
446 H. Yu et al.

Cl Cl
O
O O
O O
OH O H O
H HO H HO
O O
OH OH HO OH
O O

41 Chaetomugilin A 42 Chaetomugilin D 43 Cytosporone B

O
OH
O O
O
HO

HO OH

44 Cytosporone C 45 Brefeldin A
Fig. Antimicrobial aliphatic compouds (41-45) from endophytes
O O O

OH O OCH 3O OH O
46 47 48

O O
O HO
H3 C H
HO OH
OH OH
O CH 3
O O OCH 3 H O
O
OHC OH H3

OH
O OH O COOCH 3

49 50 sordaricin(51)
O
O

O
trichodermin(52)

Figure 3. (Continued)

against phytopathogens (Phythophtora capsici,
Phythophtora parasitica, Fusarium oxysporum and
A. solani) with IC50 values from 20 to 170 mg/mL.
The author speculated that the structure–activity
relationship reveals that the hydroxyl group at C-9
and the presence of the C-20, C-30 double bond are
possibly responsible for the higher bioactivity of
compound (38).
3.7. Aliphatic compounds
Aliphatic compounds, frequently detected in
cultures of endophytes, often show biological
activities. Two aliphatic metabolites, named chae-
tomugilin A (41) and D (42), with antifungal
activities were isolated from an endophytic
fungus C. globosum collected from Ginkgo biloba
(Qin et al. 2009). Cytosporone B (43) and C (44)
were isolated from a mangrove endophytic fungus,
Phomopsis sp. They inhibited two fungi C. albicans
and F. oxysporum with the MIC value ranging from
32 to 64 mg/mL (Huang et al. 2008). In our
laboratory we have found brefeldin A (45) synthe-
sized by an endophytic Penicillium sp. isolated
from the root of Panax ginseng. This compound is a
unique fungal metabolite of 13-membered lactone
ring, showing significant antimicrobial activity
(Wang et al. 2006).
3.8. Flavonoids
Antimicrobial flavonoids derivatives (46–48)
belonging to flavonoids were isolated from the
Recent developments and future prospects of antimicrobial metabolites produced by endophytes
culture extract of the endophytic fungus Nodulis-
porium sp. from Juniperus cedre on Gomera Island
(Dai et al. 2006). Three derivatives can serve as
lead molecules whose activities can be enhanced
by manipulation through synthetic chemistry. Fla-
vonoids have already been used to synthesize
antibiotics in some laboratories in our university.
3.9. Others
Two new cyclohexanone derivatives (49, 50),
have been isolated from cultures of the plant
endophytic fungus Pestalotiopsis fici, and both of
them showed significant antifungal activity against
Aspergillus fumigatus (Liu et al. 2009). Few
compounds, possessing antifungal activities have
been found from endophyte at present. Further
research of cyclohexanone that possesses the
character of carcinogenicity is needed to carry on
before the two metabolites applied in our lives.
Sordaricin (51) cultured by the endophytic fungus
Xylaria sp. isolated from the leaves of Garcinia
dulcis, exhibited moderate activity against a wide
range of fungal pathogens (Pongcharoen et al.
2008). Antifungal trichodermin (52) was recently
characterized from Trichoderma harzianum, an
endophytic fungus living in Llex cornuta Lindl.
Plant experimental results showed that trichoder-
min had significantly protective effect to early
blight on tomato and damping-off on cucumber
(Chen et al. 2007).
4. Conclusion and future targets
Since Stierle et al. (1993) discovered a taxol-
producing endophyte: Pestalotiopsis microspore,
there have been a increasing interests in bioactive
metabolites isolated from endophytes. Many stu-
dies proved that endophytes produce novel sec-
ondary metabolites as a resistance mechanism to
overcome pathogenic invasion (Tan and Zou 2001).
Researches of endophytes have been carried out
and a great amount of antimicrobial natural
products were found. So far, a great number of
antimicrobial compounds have been found in a
handful of the one million different endophyte
species (Petrini 1991), it is believed that searching
for natural products synthesized by endophyte
could be a promising way to solve the problem
that bacteria are becoming resistant to some
commonly used drugs (WHO 2002) and meet the
emergency demand of discovering highly effective,
low toxicity, and no environmental impacted
447
antibiotics to fight against resistant bacterial
species.
Although researches on endophytes have been
carried out for a long time, some problems should
not be ignored.
(1) The toxicities of some endophytic antimicrobial
compounds appear to be so unspecific that they
are often as toxic to other organisms including
humans as they are to fungi and bacteria;
(2) Some of the antimicrobial compounds, showing
moderate antifungal activities, cannot be used
as a potential medicine or pesticides;
(3) The yield of some antimicrobial compounds in
culture medium is too low to applied in large
scale;
(4) The biosynthesis, regulation and intermediates
of antimicrobial natural products in endophytes
are totally or partly unknown.
The ultimate purpose of endophytes research is
to find new antibiotics or pesticides, thus the
following aspects should be intensively studied:
(1) To find better bioactive antimicrobial sub-
stances without any side effect to human, plant
and environment;
(2) To enhance the antimicrobial activity or de-
crease the side effects of known metabolites
from endophytes by modifying their structures
in order to improve the efficacy and specificity
to microbes;
(3) To optimize conditions of endophyte fermenta-
tion that has been found to show bioactivity in
order to enhance the yield of active substances
synthesized by endophyte;
(4) To search for the regulatory gene in synthesis
path of antimicrobial compound, and use
genetic engineering technology to increase
the production of antibacterial substances.
As so many antimicrobial compounds were
isolated from endophytes which only occupied a
small portion of total endophyte species, it is
obviously that there is a great opportunity to find
reliable and novel antimicrobial natural products in
endophytes, which may be used as clinically
effective antibiotics in future.
Acknowledgements
The authors are grateful to the precious
comments and careful correction made by anon-
ymous reviewers. The authors also would like to
448
acknowledge the assistance of Fei You, Fang Huang
and Xiaohan Wu (SMMU) for data collection.
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