Exploring novel bioactive compounds from marine microbes

Lixin Zhang1,2,3, Rong An1,4, Jinping Wang4, Nuo Sun1,4,

Si Zhang2, Jiangchun Hu5 and Jun Kuai6

The historical paradigm of the deep ocean as a biological past, this has proven to be a rich source of extremely potent
‘desert’ has shifted to one of a ‘rainforest’ owing to the isolation compounds [3,4], which represent a considerable number
of many novel microbes and their associated bioactive of drug candidates [5]. However, to date, the biodiversity
compounds. Recently, there has been an explosion of of marine microbes and the versatility of their bioactive
information about novel bioactive compounds that have been metabolites have not been fully explored.
isolated from marine microbes in an effort to further explore the
relatively untapped marine microbes and their secondary Microbes can sense, adapt and respond to their environ-
metabolites for drug discovery. The microbes are recovered ment quickly and can compete for defense and survival
and purified from the ocean by both conventional and by the generation of unique secondary metabolites.
innovative isolation methods to obtain those previously thought These compounds are produced in response to stress
to be ‘uncultivable’. To overcome the difficulties and limitations and many have shown value in biotechnological or phar-
associated with cultivation techniques, several DNA-based maceutical applications. The marine environment was
molecular methods have been developed to bypass the once thought to have high salt, poor nutrition and less
culture-dependent bottleneck. Bioactive compounds isolated microbial growth. On the contrary, soil microbes are
using the above strategies have not only shown importance in widely regarded to live in a much more crowded and
biotechnological and pharmaceutical applications but have competitive environment. The ecology of marine natural
also increased our understanding of the diversity of marine products actually reveals that many of these compounds
microbiota, ecosystem functions and the exploitable biology. are chemical weapons and have evolved into highly
Addresses potent inhibitors of physiological processes in the prey,
1
Guangzhou Institute of Biomedicine and Health, Chinese Academy of predators or competitors of the marine organisms that
Sciences, Guangzhou 510663, China
2
utilize them for survival. Venter et al. [6] harnessed the
South China Sea Institute of Oceanology, Chinese Academy of power of high-throughput DNA sequencing and compu-
Sciences, Guangzhou 510301, China
3
SynerZ Pharmaceuticals Inc., Lexington, MA 02421, USA
tational genomics to produce a massive dataset of large
4
Department of Microbiology, Shandong University, Jinan 250100, DNA fragments from the total microbial genomes
China extracted from the subtropical North Atlantic Ocean
5
Insititute of Applied Ecology, Chinese Academy of Sciences, Shenyang off the Bermuda coast. They studied the DNA extracted
110016, China
6
Wyeth Research, Cambridge, MA 02139, USA
from 1500 l of surface seawater and within this identi-
fied more than 1.2 million new genes. The discovery of
Corresponding author: Zhang, Lixin ([email protected]) such an enormous number of new genes from such a small
sample obtained in one of the world’s most nutrient-
impoverished bodies of water poses significant challenges
Current Opinion in Microbiology 2005, 8:276–281
to the emerging field of marine molecular microbial
This review comes from a themed issue on ecology and evolutionary biology. Marine microbes live
Ecology and industrial microbiology in a biologically competitive environment with unique
Edited by Sergio Sánchez and Betty Olson conditions of pH, temperature, pressure, oxygen, light,
Available online 6th May 2005
nutrients and salinity, which is especially rich in chlorine
and bromine elements. There is no wonder that marine
1369-5274/$ – see front matter microbial metabolites exhibit special biological activities
# 2005 Elsevier Ltd. All rights reserved. compared with ‘terrestrial’ bacteria [7,8].
DOI 10.1016/j.mib.2005.04.008
The emphasis of this review is on marine microbial
producers and on how to maximize their potential to
generate novel biologically active compounds. We high-
Introduction light three different strategies to directly isolate marine
Although more than 30 000 diseases have been clinically microbes and their biosynthetic gene clusters.
described, less than one-third of these can be treated
symptomatically and only a few can be cured. New ther- Potent therapeutic agents isolated from
apeutic agents are urgently needed to treat medical needs marine organisms using a conventional
that are currently unmet. Natural products once played a culturing method
major role in drug discovery [1,2].The marine environ- Perhaps the first notable discovery of a biologically active
ment covers more than 70% of the world’s surface. In the marine natural product was reported over 50 years ago by

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 Exploring novel bioactive compounds from marine microbes Zhang et al. 277

Bergman [4]. The discovery of arabinose-based nucleo- Currently, no other approved marine drugs have been
sides was the first demonstration that naturally occurring obtained directly from the ocean; however, a significant
nucleosides could be found that contain sugars other than number of marine-derived compounds have been in clin-
ribose and deoxyribose [4]. Dedemnin B isolated from ical or in preclinical trials. More than 720 marine metabo-
the tunicate Trididemnum solidum was the first defined lites were reported in the literature during 2003, about half
marine product to enter clinical trials for any major human of which demonstrated biological activities. Yondelis, bet-
disease. Since then, there has been an explosion of ter known as ecteinascidin 743 (Figure 1), is in Phase II and
interest in alternative nucleoside compositions; the anti- III trials in Europe and in the USA, respectively, for
viral drugs Ara-C (Figure 1) and Ara-A were subsequently treatment of soft tissue sarcoma, and the Conus toxin, also
developed, with obvious linkages to later antiviral agents known as Ziconotide or Prialt, is in Phase III clinical trials
such as acyclovir and AZT (also known as zidovudine). for intractable pain [4].
Synthetic analog studies led to the development of Ara-
C (the drug name of which is cytarabine) as a clinically Table 1 summarizes the current status of representative
useful anticancer agent [4,9]. A probable biological compounds isolated from marine sources. The marine
reason for the presence of such highly bioactive com- natural products are classified in the following categories:
pounds in marine microbes could stem from the neces- antiparasitic drugs, antinematodal drugs, antituberculosis
sity of a toxin to be potent because of the diluting effect agents, antiviral leads and antifungal agents [10]. They
of seawater. However, such high bioactivity of a poten- include aureol, puupehenone, sarcophine, palinurin and
tial drug could also lead to toxicity, which is a problem manzamine alkaloids [11].
in the drug discovery process. Many drugs could be
more effective at a reduced dosage if low doses of In addition, there are many marine organisms, such as
synergistic compounds are simultaneously introduced. cone snails, whose toxic venom is used for defense and
This synergistic premise might offer a unique method contains up to 50 different peptides that selectively
for marine drug discovery and could rescue marine drug inhibit the function of ion channels involved in the
candidates that have been abandoned because of inade- transmission of nerve signals in animals [12]. As we
quate safety profiles [2]. Bryostatin (Figure 1) is likely to can see from the table, sponges, together with other
be produced by a bacterial symbiont and is now in marine organisms such as corals, are an important source
combination therapy [4]. of biologically active natural products. Several microbes
isolated from sponges and from other marine animals
produced the same compounds in the laboratory as those
Figure 1 originally isolated from their hosts. Because the majority
of marine invertebrates have no methods of physical
Salinosporamide A Ecteinascidin 743 (ET-743) defense against predators, it is reasonable to hypothesize
that symbiotic marine microorganisms are the original
HO producers of these bioactive compounds [8].
H
OH MeO NH
H OMe
N
O
O HO Me It is reported that sponges are often associated with
O
O
symbiotic microbial populations that include archaea,
Cl OAc S H
H3 C O
H bacteria, actinomycetes, fungi, cyanobacteria and micro-
CH3
N algae. In some cases, it is these microorganisms, and not
N the sponge cells, that are the probable source of the
O H
O OH secondary metabolites of interest [13]. For example,
the polybrominated biphenyl ether antibiotics isolated
from the sponge Dysidea herbacea are really produced by
Ara-C (Cytarabine) Bryostatin 1 the endosymbiotic cyanobacterium Oscillatoria spongeliae
OAc
[13]. Work on isolation of and cultivation of sponge
O H OH
H MeO2C symbionts and the nature of their symbiotic relationships
N OHO O
H2N N OH
have been reviewed elsewhere [13]. Not surprisingly,
O O many marine microbes have been cultivated and isolated
H
OH OH H
O OHO from seawater, sediments and symbiotic hosts by con-
ventional approaches inherited from soil microbiology
HO
[14]. In addition to the open ocean, there are diverse
CH3(CH2)2(CH)4CO2
and dynamic areas, such as mangroves, coral reefs, hydro-
CO2Me thermal vents and deep-sea sediments, in which microbes
Current Opinion in Microbiology can be searched for. Natural products have been isolated
from marine invertebrates such as sponges, tunicates,
Structures of some representative marine bioactive metabolites. mollusks and bryozoans. This not only demonstrates

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278 Ecology and industrial microbiology

Table 1

Representative marine metabolites with different therapeutic activities.

Compound Type Source

Antifungal activity
Aurantoside B Polyketide Siliquariaspongia japonica sponge
Phorboxazole A Macrolide Phorbas sp. sponge
Halishigamide A Macrolide Halichondria sp. sponge
Halichondramide Macrolide Halichondria sp. sponge
Fascaplysin Bis (indole) alkaloid Fascaplysinopsis sp. sponge
Meridine Polycyclic alkaloid Corticium sp. sponge
Bengazole A Oxazole-containing fatty-acid ester Jaspis sp. sponge
Ptilomycalin A Polycyclic Ptilocaulis spiculifer guanidine alkaloid sponge
Antituberculosis activity
Alkaloid (+)-8-hydroxymanzamine Heterocyclic ring system attached Pachypellina sp. sponge and Petrosiidae genus
to a-carboline moiety
Axisonitrile-3 Cyanosesquiterpene Acanthella klethra sponge
Litosterol C-19 hydroxysteroid Okinawan soft coral Litophyton viridis
Puupehenones Shikimate-sesquiterpene Sponges
Antiviral activity
Papuamides A Cyclic depsipeptides Theonella mirabilis and T. swinhoei sponges
Avarone Sesquiterpene hydroquinone Dysidea avara sponge
Gymnochrome D Brominated phenanthroperylenequinone Fossil crinoid Gymnocrinus richeri pigments
Microspinosamide Cyclic depsipeptide Sidonops microspinosa sponge
Solenolide A Diterpene lactone Gorgonian of the genus Solenopodium
Hennoxazole A Oxazole-containing alkaloid Polyfibrospongia sp. sponge
Thyrsiferol Triterpene Red alga Laurencia venusta
Spongiadiol Tetracyclic furanoditerpene Deep-water Spongia sp
Antibacterial activity
Squalamine Aminosterol Dogfish shark Squalus acanthias
Cribrostatin 3 Isoquinolines Cribrochalina sp. sponge
Bromosphaerone Bromoditerpenes Sphaerococcus coronopifolius red alga
Pestalone Chlorinated benzophenone Rosenvingea sp. brown alga
Jorumycin Dimeric isoquinoline alkaloid Jorunna funebris
Antiprotozoal activity
Cyclic peroxides Peroxide Plakortis aff angulospiculatus sponge
Sigmosceptrellin-B Alkaloid Diacarnus erythraeanus red sea sponge
Plakortide Cyclic peroxylactone Plakinastrella onkodes sponge
Ascochyta salicorniae Fungus Ulva sp. green alga
Anthelmintic activity
Dihydroxytetrahydrofuran Tetrahydrofuran Notheia anomala south Australian
marine brown alga
Amphilactams Amphimedon sp. sponge
Geodin A magnesium salt Macrocyclic polyketide Geodia sp. sponge
lactam tetramic acid

the numerous opportunities that oceans provide for the Salinospora sp. that originates from a heat-treated marine
discovery of new compounds but also validates the phar- sediment sample in the Bahamas. The structure of sal-
macological value of the exploration of oceans for novel inosporamide A, including its absolute stereochemistry,
compounds. was deduced by way of spectral and X-ray analyses.
Salinosporamide A displayed potent and selective in vitro
There are some concerns about the isolation of marine cytotoxicity against cell lines in the National Cancer
microbes. Some researchers claim that these organisms Institue (NCI) panel. It also exhibited inhibition of the
are hard to maintain in the laboratory environment. chymotrypsin-like proteolytic activity of the purified 20S
However, one successful case was the recent discovery proteasome. The unique function of the core-fused-
of a new genus of actinomycetes, Salinospora, which can lactam–lactone bicyclic ring structure of salinosporamide
only be found in the marine environment [15,16]. In A appears to contribute to its anti-cancer potency. It is
excess of 2,500 strains of this taxon have now been therefore not surprising that this compound showed potent
isolated. The potent proteasome inhibitor salinospora- inhibition of cancer growth, including human colon carci-
mide A (Figure 1) was isolated from a culture of a noma, non-small cell lung cancer and breast cancer.

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 Exploring novel bioactive compounds from marine microbes Zhang et al. 279

It seems probable that there are many natural products signaling molecules that are produced by other micro-
awaiting to be discovered and tested for different medical organisms. It enables cells to grow with nutrients that are
applications. Indeed, more and more novel marine present in their natural environment. After long periods of
microbes and novel bioactive secondary metabolites will incubation with a constant flow of a very low concentra-
continue to be isolated [7]. tion of nutrients, flow cytometry was used to separate gel
beads containing cells that had grown into small micro-
Innovative ways to isolate less-culturable or colonies from those that had not. Many novel marine
uncultivable marine microbes bacteria have been isolated in this way, including mem-
Microbial community analyses by epi-fluorescence micro- bers of previously uncultured groups that are abundant in
scopy and rRNA sequencing strategies have revealed that seawater [21,22,23].
the microorganisms that can be cultured only represent a
small fraction of the microbial population and might not be One argument against the application of less-culturable
the most prevalent in the natural environment. It is expec- strains to a drug discovery program is that they can not be
ted that one of the largest efforts in this field of research in cultured at a high cell density. The counter argument is
the next decade will be the exploration of a means to grow that, although they might initially require the addition of
less-culturable marine microbes. It is thought that the growth factors or oligotrophic growth conditions, there is
reason for the enormous discrepancy between the total evidence that once cultured the organisms can be grown
viable cell count and the cell count of culturable cells might using nutrient-rich media. Using 960 cells cultured in
be because of the following: cell damage by oxidative microdroplets, 67% of the cultures were subsequently
stress; formation of viable but non-culturable cells; able to grow to densities of >107 cells per ml [21,22,23].
inhibition by high substrate concentrations; induction of This allows the cells to be cultured in a manner that could
lysogenic phages upon starvation; and lack of cell–cell easily be applied to drug discovery platforms.
communication in laboratory media. Among the appro-
aches used to enhance the resuscitation of less culturable The use of metagenomic methods to bypass
strains, are the addition of cell signaling molecules and the the culture-dependent bottleneck
use of oligotrophic isolation media [17,18]. It is widely accepted that more than 99.8% of the microbes
present in many environments are not readily culturable.
One method utilized the concept of ‘extinction culturing’ Metagenomic technology tries to overcome this bottleneck
to isolate cultures in small volumes of low-nutrient media by the development and the utilization of culture-inde-
(typically three orders of magnitude less than common pendent approaches. The major sequencing approach in
laboratory media). Many novel strains among the Proteo- the Sargasso Sea identified a number of DNA sequences
bacteria, Planctomycetes, Bacteroidetes, Acidobacteria and Ver- that was by far greater than had previously been detected
rucomicrobia were obtained; these included some [6]. This study is likely to lead to new investigations of
previously believed to be uncultivable [18]. Several novel the microbial world and its evolution. A recent example
bacterial strains from the western Sargasso Sea were gave significant insights into the community structure and
successfully isolated using this method [19]. the metabolism of a natural acidophilic biofilm that is
growing on the surface of drainage flowing from an acid-
Another development is based on the assumption that mine. This was mainly possible through the reconstruction
uncultivable microorganisms might grow in pure culture of microbial genomes that are present in this niche. For this
if provided with the chemical components of their natural purpose, the near-complete genomes of Leptospirillum
environments. To test this hypothesis, specialized growth group II and Ferroplasma type II were reconstructed. A
chambers were developed that mimic the natural marine detailed analysis of these genomes allowed pathway-recon-
environment of the microorganisms to optimize the iso- struction of carbon fixation and energy generation and
lation of pure cultures in vitro. Microorganisms in sedi- provided insights into the survival strategies of microbes
ment samples were serially diluted with seawater, mixed within this extreme acidic environment [24]. Cultivation-
with agar, placed in diffusion chambers, and incubated in independent biochemical studies of microbial mats of a
an aquarium that simulated the environmental conditions northwestern Black Sea shelf, which oxidize methane
of the sampling site. The number of microcolonies under anaerobic conditions, resulted in the identification
obtained was in the range of 2–40% of the cells inocu- of a prominent nickel-containing protein. This approach is
lated, which is much higher than the recovery rate for not only remarkable because of the biochemical and eco-
conventional cultivation methods [20]. logical findings but also because it uncovered, for the first
time, the function of a novel protein. Metagenomic tech-
A high throughput cultivation method based on the nologies were subsequently applied to determine the
combination of single cell encapsulation and a gel micro- corresponding gene [25].
droplet was also developed. Gel microdroplets are small
porous gel matrixes that encapsulate individual cells. The metagenomic approach maximizes the diversity of
This method allows the exchange of metabolites and/or libraries of marine natural product extracts by accessing

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280 Ecology and industrial microbiology

the DNA directly from marine samples [23,26]. Such been isolated from marine microbes. It is therefore timely
DNA can be directly isolated, digested into large frag- to review the past successes of marine microbial natural
ments with restriction enzymes, and cloned into an arti- products as medicines and to examine future possibilities
ficial vector. The vector is then transformed into a that arise from both conventional and new technologies to
surrogate host. Environmental DNA libraries can be further explore the biodiversity of marine microbes and
prepared that contain large fragments of DNA from a their associated secondary metabolites.
wide range of uncultivated bacteria within an environ-
mental sample. This recombinant approach obviates the However, future success is not a matter of old versus new;
need for culturing diverse microorganisms and provides a it is dependent on learning how to apply the existing
relatively unbiased sampling of the vastly untapped methodologies of genomics, proteomics, combinatorial
genetic diversity that is present in various microenviron- chemistry, DNA shuffling, combinatorial biosynthesis,
ments. As an additional advantage, the genes that encode biodiversity, bioinformatics and high-throughput screen-
biosynthesis of a product of interest can be isolated and ing to rapidly evaluate the activities in extracts as well as
analyzed using the bioinformatics tools, thus providing a in purified components derived from marine microbes.
potential boost to the efforts of analytical chemists to The remarkable chemical diversity encompassed by nat-
identify the product. Furthermore, the possibility of ural products continues to be of relevance to drug dis-
regulating the expression of isolated environmental gene covery. Although today’s drug discovery engine operates
clusters, or combining them with genes for other path- at an accelerated pace compared with the era in which
ways to obtain new compounds, furnishes a further advan- natural products were pre-eminent sources of drug leads,
tage over traditional natural product discovery numerous approaches have been developed to capture
methodologies. However, it must be noted that these the intrinsic value of their drug properties. Biodiversity,
biosynthetic and regulatory genes might be dormant in creativity and versatility of marine microbial metabolites
the host and that optimal induction conditions might be can now be studied by way of culture-dependant and -
required for the production of novel natural products [14]. independent methods. We are in a position to start
The screening of metagenomic libraries has resulted in probing many major questions that, only a few years
the identification of a significant number of novel genes ago, were beyond our grasp, among them being the
that encode for biocatalysts or for molecules that have a functional role of these microbes and their metabolites
high potential use in pharmaceutical products or in pro- in marine habitats as well as the significance of ecological
duction processes. The isolation of genes that encode for and evolutionary processes of such high microbial diver-
novel therapeutic molecules is a valuable area of research. sity in the sea.
The genes of interest within these searches are often type I
and type II polyketide synthases (e.g. b-ketoacyl synthe- Acknowledgements
tases), which are key genes involved in the synthesis of We thank Marcia S Osburne, Guochun Zhou and Arnold L Demain for
critical reading and for helpful discussions.
polyketide antibiotics and antitumor agents by Gram-
positive Streptomyces and are often part of large biosynthetic
References and recommended reading
gene clusters [27]. Papers of particular interest, published within the annual period of
review, have been highlighted as:
Advances in DNA sequencing and in bioinformatic tech-  of special interest
nologies now make it possible to rapidly identify the  of outstanding interest
clusters of genes that encode bioactive compounds and
to make computer predictions of their chemical structure 1. Demain AL, Zhang L: Natural products and drug discovery. In
 Natural Products: Drug Discovery and Therapeutics Medicines.
based on gene-sequence information [28,29]. These Edited by Zhang L, Demain A. Humana Press; 2005: 3-32.
structure predictions can be used to identify new chemi- This is a historical review of the great contribution that microbial natural
products have made to drug discovery.
cal entities before they are expressed in diversified fer-
mentation broth and provide important physicochemical 2. Zhang L: Integrated approaches for discovering novel drugs
from microbial natural products. In Natural Products: Drug
‘‘handles’’ that guide compound purification and struc- Discovery and Therapeutics Medicines. Edited by Zhang L,
ture confirmation. Genome sequence tags (GSTs) are Demain A. Humana Press; 2005:33-56.
genes involved in natural product biosynthesis. These 3. Gochfeld DJ, El Sayed KA, Yousaf M, Hu JF, Bartyzel P,
GSTs are used as probes to screen for the presence of Dunbar DC, Wilkins SP, Zjawiony JK, Schinazi RF, Schlueter Wirtz
S et al.: Marine natural products as lead anti-HIV agents.
such genes within a clonal library. Any clone that contains Mini Rev Med Chem 2003, 3:401-424.
a GST can then be screened for novel natural-product
4. Newman DJ, Cragg GM: The discovery of anticancer drugs
gene clusters. More than 450 natural-product clusters  from natural sources. In Natural Products: Drug Discovery and
have been identified in this manner [29]. Therapeutics Medicines. Edited by Zhang L, Demain A. Humana
Press; 2005:275-294.
This includes numerous, very interesting, molecules that have come from
Conclusions marine sources, or that have been synthesized as a result of knowledge
gained from prototypical compounds. Many are either in or approaching
The past three years have seen an explosion of informa- Phase II or III clinical trials for cancer, analgesia, allergy and cognitive
tion in the field of novel bioactive compounds that have disease. A substantial number of other potential agents is also provided.

Current Opinion in Microbiology 2005, 8:276–281 www.sciencedirect.com

 Exploring novel bioactive compounds from marine microbes Zhang et al. 281

5. Haefner B: Drugs from the deep: marine natural products as heterotrophic bacteria from the central Baltic Sea.
 drug candidates. Drug Discov Today 2003, 8:536-544. Appl Environ Microbiol 2002, 68:3978-3987.
This work is very interesting because it reviews a variety of marine natural
products as drug candidates that have been discovered over a long 18. Connon SA, Giovannoni SJ: High-throughput methods for
period of time in the angle of drug efficacy and specificity for the treatment culturing microorganisms in very-low-nutrient media yield
of many human diseases. diverse new marine isolates. Appl Environ Microbiol 2002,
68:3878-3885.
6. Venter JC, Remington K, Heidelberg JF, Halpern AL, Rusch D,
 Eisen JA, Wu D, Paulsen I, Nelson KE, Nelson W et al.: 19. Cho JC, Giovannoni SJ: Parvularcula bermudensis gen.
Environmental genome shotgun sequencing of the Sargasso nov., sp. nov., a marine bacterium that forms a deep branch
Sea. Science 2004, 304:66-74. in the a-Proteobacteria. Int J Syst Evol Microbiol 2003,
This publication is remarkable because of the large number of novel genes 53:1031-1036.
detected within the enormous number of DNA sequences generated.
20. Kaeberlein T, Lewis K, Epstein SS: Isolating ‘uncultivable’
7. Blunt JW, Copp BR, Munro MH, Northcote PT, Prinsep MR: microorganisms in pure culture in a simulated natural
 Marine natural products. Nat Prod Rep 2004, 21:1-49. environment. Science 2002, 296:1127-1129.
This review covers the literature published in 2003 regarding marine
21. Keller M, Zengler K: Tapping into microbial diversity. Nat Rev
natural products. It contains 619 citations.
 Microbiol 2004, 2:141-150.
8. Berdy J: Bioactive microbial metabolites. J Antibiot (Tokyo) This excellent review tapped into the recombinant DNA technology and a
2005, 58:1-26. large number of new enzymes by cloning genes directly from the envir-
onment using environmental clone libraries, thereby accessing the tre-
9. Baker DD, Alvi KA: Small-molecule natural products: new mendous microbial diversity of uncultivated microorganisms.
 structures, new activities. Curr Opin Biotechnol 2004, 15:576-583.
This is a good review article that summarizes the trends in natural product 22. Zengler K, Toldeo G, Rappe M, Elkins J, Mathur EJ, Short JM,
discovery and that highlights some of the recent discoveries of novel Keller M: Cultivating the uncultured. Proc Natl Acad Sci USA
structures and interesting activities published from 2002 to 2004. 2002, 99:15681-15686.

10. Donia M, Hamann MT: Marine natural products and their 23. Zengler K, Paradkar A, Keller M: New methods to access
 potential applications as anti-infective agents. Lancet Infect Dis microbial diversity for small molecule discovery. In Natural
2003, 3:338-348. Products: Drug Discovery and Therapeutics Medicines. Edited by
This review focuses on the pharmacologically tested marine leads that Zhang L, Demain A. Humana Press; 2005:275-294.
have shown in vivo efficacy or potent in vitro activity against infectious
24. Tyson GW, Chapman J, Hugenholtz P, Allen EE, Ram RJ,
and parasitic diseases.
 Richardson PM, Solovyev VV, Rubin EM, Rokhsar DS, Banfield JF:
11. Cooper EL: Commentary on ‘Traditional and modern Community structure and metabolism through reconstruction
 biomedical prospecting: part II — the benefits’ by Müller WEG, of microbial genomes from the environment. Nature 2004,
Schröder HC, Wiens M, Ottstadt SP, Batel R and Müller IM. 428:37-43.
Anti-protozoa and antiviral activities of non-cytotoxic This is one of the first publications to present a near-complete set
truncated and variant analogues of mussel defensin by Roch P of metagenomic sequences from a mixed but low-diversity microbial
Beschin A and Bernard E. eCAM 2004, 1:207-209. community.
This focuses on the cited papers and comments on the application, the
25. Kruger M, Meyerdierks A, Glockner FO, Amann R, Widdel F,
resource and the efficacy of marine natural products as drug candidates.
 Kube M, Reinhardt R, Kahnt J, Bocher R, Thauer RK et al.: A
12. Livett BG, Gayler KR, Khalil Z: Drugs from the sea: conopeptides conspicuous nickel protein in microbial mats that oxidize
 as potential therapeutics. Curr Med Chem 2004, 11:1715-1723. methane anaerobically. Nature 2003, 426:878-881.
This review describes the progress made by several research groups to This publication is remarkable as it identifies the functional role of a novel
characterize the properties of conopeptides and to use them as drug protein involved in the global carbon (methane) cycle and it subsequently
leads for the development of novel therapeutics for the treatment of a identifies its DNA sequence.
range of neurological conditions.
26. Martinez A, Hopke J, MacNeil IA, Osburne MS: Accessing
13. Belarbi El H, Gomez AC, Chisti Y: Producing drugs from marine the genomes of uncultivated microbes for novel
sponges. Biotechnol Adv 2003, 21:585-598. natural products. In Natural Products: Drug Discovery and
Therapeutics Medicines. Edited by Zhang L, Demain A. Humana
14. Knight V, Sanglier JJ, DiTullio D, Braccili S, Bonner P, Waters J, Press; 2005:295-314.
Hughes D, Zhang L: Diversifying microbial natural products for
drug discovery. Appl Microbiol Biotechnol 2003, 62:446-458. 27. Streit WR, Schmitz RA: Metagenomics — the key to the
 uncultured microbes. Curr Opin Microbiol 2004, 7:492-498.
15. Mincer TJ, Jensen PR, Kauffman CA, Fenical W: Widespread and This paper is a remarkable overview of the recent progress in metagen-
persistent populations of a major new marine actinomycete ome technology.
taxon in ocean sediments. Appl Environ Microbiol 2002,
68:5005-5011. 28. Zazopoulos E, Huang K, Staffa A, Liu W, Bachmann BO, Nonaka K,
Ahlert J, Thorson JS, Shen B, Farnet CM: A genomics-guided
16. Jensen PR, Fenical W: New natural-product diversity from approach for discovering and expressing cryptic metabolic
marine actinomycetes. In Natural Products: Drug Discovery and pathways. Nat Biotechnol 2003, 21:187-190.
Therapeutics Medicines. Edited by Zhang L, Demain A. Humana
Press; 2005:315-328. 29. Farnet CM, Zazopoulos E: Improving drug discovery from
microorganisms. In Natural Products: Drug Discovery and
17. Bruns A, Cypionka H, Overmann J: Cyclic AMP and acyl Therapeutics Medicines. Edited by Zhang L, Demain A. Humana
homoserine lactones increase the cultivation efficiency of Press; 2005:95-106.

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