Jensen. 1994, Strategies For The Discovery of Secondary Metabolites PDF

ANNUAL
REVIEWS Further
Annu. Rev. Microbiol. /994. 48:559-84 Quick links to online content
Copyright Ii) 1994 by Annuol Reviews Inc. All rights reserved
STRATEGIES FOR THE

DISCOVERY OF SECONDARY
METABOLITES FROM MARINE
BACTERIA: Ecological
Annu. Rev. Microbiol. 1994.48:559-584. Downloaded from www.annualreviews.org
Perspectiyes
by University of Wisconsin - Madison on 08/07/12. For personal use only.
Paul R. Jensen and William Fenical

Scripps Institution of Oceanography, La Jolla, California 92093-0236
KEY WORDS: bacterial ecology, biological activity, marine natural products chemistry
CONTENTS
INTRODUCTION . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 560
ECOLOGICAL CONSIDERATIONS ...... .... . .. . . ... . . .... . .. . . . .. . . .. . . . . 561

Prokaryotic Diversity. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 563
The Distribution of Bacteria in the Sea .. ... . . . ......... . ... . . . . .. . . .. . .. . . 564
The Associations of Bacteria with Plants and Invertebrates . . . . . .. . . . . . . . . . .. . . 566
The Role of Symbiotic Bacteria ill 567
Effects of Marille Metabolites UpOIl the Distributioll of Bacteria . . .. . .. .... . .. . . 569
The Adaptations of Bacteria to Diverse Marill 570
Bacterial Culturability. . . . . . ..... . .. . .. . . . .. . . ... . . . . .. . . .. . . .. . . .. . .. . . 571
The Distinctioll Betweell Marine alld Terrestrial Bacteria . ......... .... ..... . . 572
BIOLOGICAL ACTIVITIES AND NOVEL ORGANIC SUBSTANCES............ 574
CONCLUDING REMARKS. . . . . . . . .. . ..... . . .. . . . .. . . . .... . . . . . . . . . . . . .. . . 576
ABSTRACf
Marine microorganisms have become an important point of study in the search

for novel microbial products. Today, both academic and industrial interest in
marine microorganisms is on the rise, in part because of the growing number
of unique, biologically active secondary metabolites reported from marine
bacteria. OUf ability to assess the biosynthetic potential of marine bacteria,
however, is inevitably coupled to our basic understanding of their biology.
This review summarizes the chemical discoveries and biological activities
559
0066-4227/9411001-0559$05.00
560 JENSEN & FENICAL
reported from marine bacteria and focuses upon several microbiological topics
as they relate to natural product discovery, including the distributions, diver
sity, and culturability of marine bacteria, as well as the role of symbiotic
bacteria in the production of substances previously ascribed to other sources.
INTRODUCTION
Historically, the study of natural products has focused upon the elaborate
biosynthetic pathways of terrestrial plants and microorganisms. In the late

1960s, however, the search for novel metabolites took a new direction as the
realm of exploration expanded to include plants and animals in the sea. This
avenue of research was initiated primarily by academicians and facilitated by
the development of scuba, which provided an effective means to collect shal
low-water marine organisms. Today, as the result of extensive research, the
study of marine natural products is recognized as both an integral component
of natural products chemistry and a significant resource for drug discovery. In
addition, the successes of marine natural products chemistry have helped
nurture the growing discipline of marine chemical ecology, an area of research
that has contributed significantly to our understanding of the ecological roles
of marine secondary metabolites (see 142).
The discovery that marine organisms represent a rich source of previously
undescribed secondary metabolites is not entirely surprising considering that
many algal and invertebrate phyla reside exclusively in the sea. In hindsight,
this discovery seems only logical given that many soft-bodied marine organ
isms, lacking obvious structural defenses, produce toxic secondary metabolites
as a means of chemical defense. Although our understanding of chemical
defense and other ecological roles of marine secondary metabolites has just
begun, chemical studies of marine organisms have been extensive, yielding
thousands of unique metabolites featuring new structural classes, functional
groups, and a relatively high incidence of halogenation. Descriptions of the
chemical discoveries in this field can be found in periodically updated reviews
from 1977 to date (36, and previous reviews cited therein).
As the chemical prolificacy of marine organisms became apparent, so did
their industrial potential, and marine metabolites are now being screened
extensively for pharmacological activity. The results of these efforts have led
to several significant biomedical discoveries. These include the clinical evalu
ation of at least two marine natural products as anticancer agents-bryostatin
I (143,167), which is currently in clinical trials in both Great Britain and the
United States, and didemnin B (150), which recently completed phase I clinical
trials in the United States (see 77). For descriptions of the biological activities
of various marine plant and invertebrate taxa (151), as well as detailed reviews
SECONDARY METABOLITES FROM MARINE B ACTERIA 561
of the biomedical potential and pharmacological properties of marine metabo

lites, a number of reviews are available (3, 38).
Continuing efforts to obtain chemically unique marine organisms have led
to the use of new and innovative methods of sample collection. These efforts
include travel to far reaches of the globe to collect previously unstudied
species, the use of deep-diving submersibles to access habitats that lie beyond
the depths of conventional scuba, and the collection of organisms that were
previously overlooked owing to difficulties in obtaining adequate biomass for
chemical evaluation. These efforts have accompanied a growing interest in the
study of marine bacteria, a logical and potentially productive avenue of re

search given the historical significance of secondary metabolites from terres
trial bacteria and the renewable nature of culturable resources.

During the past decade, studies of secondary metabolites from marine bac
teria have begun in laboratories throughout the world. In the most simplified
sense, these studies use standard dilution and plating techniques to isolate
bacteria from marine samples. Isolated bacteria are cultured in shake-flasks,
on agar plates, or in fermentors, and the organic components of these cultures
are then extracted, usually with organic solvents. The extracts are tested for
biological activities, e.g. standard antibiotic disc diffusion assays, and analyzed
for the presence of unusual chemical constituents. Organic materials of interest
are purified and their structures analyzed, a complex process that generally
requires sophisticated NMR methodologies.
Because of growing interest in the study of secondary metabolites from
marine bacteria, we would like to address in this review several topics in
microbial ecology that are relevant to natural product discovery. As these topics
are broad in scope and fundamental to marine microbiology, our goal is to
focus upon the aspects that pertain to natural product discovery. Rather than
discuss in detail chemical discoveries from marine bacteria, we refer the reader
to recent reviews oriented toward this objective (5, 39, 40, 87, 131) and offer
a summary of the biological activities and novel secondary metabolites re
ported to date from marine bacteria. All of the following discussions of marine
bacteria exclude cyanobacteria (unless otherwisc indicated), which are none
theless an important resource for unique secondary metabolites (36, and pre
vious reviews cited therein). In addition, one should note that novel secondary
metabolites have been reported from other taxonomic groups of marine mi
croorganisms, including fungi and microalgae; however, these groups are not
discussed here.
ECOLOGICAL CONSIDERATIONS
By extending the search for novel microbial metabolites to the sea, a funda
mental question must be addressed, i.e. do marine bacteria produce previously
Table 1 Novel organic molecules reported from marine bacteria
Producing strain Source Biomedical activity References
Actinomycete
Chaina purpurogena Sediment Antibiotic, anticancer 86, 136

Streptomyces tenjimariensis Sediment Antibiotic 67, 132
Streptomyces griseus Sediment Antibiotic 120, 135, 157
Streptomyces sioyaensis Sediment Anticancer 1 68, 169
Maduromycete Sediment Antibiotic 141
Streptomyces sp. Sediment Antibiotic 140
Actinomycete Sediment Antibiotic 139
Actinomycete Sediment No activity reported 138

Actinomycete Gorgonian soft coral Anticancer, antibiotic 173

Streptomyces sp. Jellyfish Antibiotic, antiinflammatory 1 79
Streptomyces sp. Sponge Antibiotic 76
Nonactinomycete
Pseudomonas bromoutilis (Alteromo- Seagrass Antibiotic 18. 107

nas sp.)
Chromobacterium (Altermonas) sp. Seawater Antibiotic 2
Pseudomonad Tide pool seawater Antibiotic 198
Beneckea gazogenes (Vibrio) Salt marsh sediment No activity reported 49
Altermonas ruba Not reported Bronchodilator 65
Unidentified Seawater No activity reported 121, 122
Unidentified gram-positive species Deep-sea sediment Anticancer, antiviral 58
Alteromonas sp. Sponge Anticancer 159
Alteromonas haloplanktis Deep-sea sediment Anticancer 85, 170
Vibrio anguillarum Fish No activity reported 79
Alteromonas sp. Seawater Enzyme inhibition 59
Alteromonas sp. Sponge Antimicrobial 166
Pseudomonas sp. Anticancer 1 23
Pseudomonas flourescens Ascidian Antimicrobial 124
Unidentified gram-positive species Deep-sea sediment Anticancer 26
Thermococcus Hydrothermal Antifungel 149, 1 52
Bacillus Deep-sea sediment Anticancer 178
Alteromonas luteoviolacea Seawater No activity reported 149
undescribed s econdary metabolites, and if so, are environmental factors spe

cific to the ocean required for the production of these substances? This question
is reminiscent of one fundamental to the development of marine microbiology
as a scientific discipline: that is, do marine bacteria represent unique species
or merely terrestrial strains adapted to the conditions of the sea?
In response to the latter question, the existence of specific marine bacteria
was documented nearly 50 y ears ago (see 108). The ability of marine bacteria
to produce previously undescribed secondary metabolites has also been ade
quately established (Table 1). Concerning the conditions necessary for the
production of secondary metabolites by marine bacteria,the isolation of a novel
metabolite does not in itself preclude the biosynthesis of this same substance
SECONDARY METABOLITES FROM MARINE BACTERIA 563
by a nonmarine strain. However, if this substance is produced only when

conditions specific to the marine environment are provided, e.g. seawater,
increased hydrostatic pressure, or a marine nutrient, the likelihood of this
substance being produced as a nonmarine fermentation product is low. More
over, if this substance possesses features rarely observed with metabolites from
terrestrial bacteria, such as the presence of halogen atoms, the argument for
its unique marine nature is further strengthened.
From a natural products perspective, marine bacteria remain a relatively
unexplored resource for novel secondary metabolites. Further investment in
this area, from both an academic and industrial perspective, will ultimately
determine the utility of this resource. To maximize the effectiveness of future
research, a better understanding of marine bacterial ecology is essential. This

understanding can be obtained through continued studies in, for example, the
diversity, distribution, and culturability of marine bacteria, and the adaptations
of bacteria for survival in both normal and extreme marine environments.
Prokaryotic Diversity
Bacterial taxonomic and metabolic diversity are fundamentally interrelated

and therefore important to natural product discovery. In marine systems, how
ever, we are only beginning to understand the true extent of prokaryotic
diversity. This fact is adequately demonstrated by the discoveries of abundant
prochlorophytes (22), cyanobacteria (189), and most recently, archaea (28, 42)
inhabiting the world's open oceans. These discoveries, in part accounted for
by the application of techniques in molecular biology to the study of microbial
ecology, confirm previous beliefs that bacterial diversity in marine systems is
far greater than can be accounted for by the culture-dependent techniques
typically used to describe bacterial communities. As studies of prokaryotic
diversity continue, the knowledge gained can be used to assess the degree to
which diverse members of the marine prokaryotic community can be obtained
in culture and included in the search for novel microbial products.
The observation that taxonomically diverse prokaryotes have not been given
adequate consideration in chemical studies of marine bacteria is supported by
the relatively few genera from which novel molecules have been reported
(Table 1). Are these the only taxa that produce unique metabolites, or are they
simply the only taxa studied to date? Most research in this field seems to have
concentrated upon marine-derived actinomycetes and gram-negative, het
erotrophic bacteria capable of growth on nutrient-rich media. This approach
is logical as the isolation and culture techniques used for these bacteria are
relatively simple and many strains can be obtained with minimum effort.
Moreover, because studies of marine bacteria have not been extensive, the
strains isolated will likely be ncw to natural products research. This approach,
however, neglects the possibility that other diverse taxa produce novel secon
dary metabolites.
Expanding natural products studies to i nclude diverse bacterial groups (e.g.
anaerobes) requires specific microbiological expertise and presents the oppor
tu ni ty for collaborations between natural products chemists and microbiolo
gists familiar with these methodologies. The challenges of establishing a
microbial screening program, especially one that includes diverse bacterial
taxa, are well known to the pharmaceutical i ndustry,and the approaches used
to meet these challenges have been described (e.g. 12,19,29,148). Although
a correlation between taxonomic and biosynthetic diversity has yet to be

defined for marine microorganisms, the inclusion of diverse bacterial taxa in
natural products research still represents a rational approach to novel metabo

lite discovery.
The Distribution of Bacteria in the Sea

The factors controlling the distribution of bacteria in the sea are complex and
vastly different from those experienced by terrestrial bacteria. The most pro
found of these differences may well be the aqueous milieu itself, which pro
vides an effective medium for ba cteri al dispersa l. Considering that seawater
typically contains 104 to 106 bacteria ml-1 (4), and ocean currents can transport
lhese organisms considerable distances, marine bacteria may encounter a far
greater variety of ha bitats and environmental conditions than terrestrial bacte
ria. The ability of marine bacteria to surviv e in and utilize the resources found
in these various habitats will ultimately have a significant effect upc;m their
distribution in the sea.
The habitats that marine bacteria encounter include surfaces, both animate
and inanimate. In contrast to terrestrial systems, marine surfaces are c onti nually
bathed by an assemblage of bacteria. The suitability of these various surfaces
for colonization may significantly affect bacterial distributions in marine habi
tats. Although the numbers and types of bacteria colonizing marine surfaces
have not been adequately documented, the colonization process and the factors
affecting colonization have been discussed in detail (24, 41,101, 117, 187).
From the perspective of natural product discovery, a general understanding
of the distribution of marine bacteria and the factors affecting these distribu
tions is beneficial. If a specific taxonomic group of bacteria is chosen for
chemical study, i nformation describing the distribution of that group will
facilitate the collection of appropriate samples. In general,these samples may
inc lud e seawater, sediments, and both animate and inanimate objects [S ie burt h
(161) has reviewed the associations of bacteria with marine samples]. Consid
ering that the distri bution of bacteria is not uniform throughout the marine
environment, qualitative descriptions of these distributions can be used to
maximize the number of diverse bacterial taxa obtained in culture.
SECONDARY METABOLlTES FROM MARINE BACTERIA 565
In the course of isolating marine bacteria for chemical evaluation, we have

surveyed the distributions of one chemically significant group, the actinomy
cetes. The best marine source of actinomycetes identified thus far are sedi
ments, from which their isolation is well documented (8, 5 3, 63, 82, 144, 1 45,
1 72, 185, 1 92). The relative numbers of actinomycetes in marine sediments
are low, however, and selective isolation methods (e.g. 34, 54, 69, 99, 1 12,
1 26), which include heat and antibiotic treatment, are useful. Following iso
lation, methods suitable for the culture and chemical evaluation of marine
actinomycetes include those used historically by the pharmaceutical industry
for soil actinomycetes, with the most common difference being the incorpo
ration of seawater or specific marine nutrients into the growth medium (e.g.
1 36).
The value of studying actinomycete distributions in sediments is demon
strated by the observation that their numbers can vary depending upon the
depth from which samples are collected ( 1 91, 1 92). For example, in a study
of near-shore marine sediments, streptomycetes were encountered infrequently
past the sublittoral zone, while actinoplanetes were isolated in greater numbers
as the distance from shore increased (82). This information is not only useful
when targeting specific taxa for isolation but can be used to assess the indige
nous nature of actinomycete isolates-an important consideration, as the origin
and activity of actinomycetes isolated from marine sources has been questioned
(53, 56, 137). These questions are valid given that actinomycetes are more
abundant in terrestrial soils than in marine sediments (55), show varying
degrees of salt tolerance (98, 1 34, 1 37, 1 77), and produce spores that undoubt
edly wash in large numbers from shore into the sea where they may remain
viable for an undetermined period of time.
Although questions about the indigenous marine nature of actinomycetes
are difficult to address experimentally, some progress has been made in this
regard. For example, we have observed that the degree to which marine-derived
actinomycetes require saline conditions for growth varies depending upon the
taxonomic group studied (82). In this example, virtually all sediment-derived
actinoplanetes required seawater for growth and were found in greater numbers
as distances from shore increased. These observations suggest that the acti
noplanetes were highly adapted to the marine environment and represent
obligate marine bacteria. On the other hand, the number of streptomycetes
decreased as distances from shore increased. These bacteria could grow in a
nonmarine medium, which indicates that the streptomycetes are facultatively
marine and possibly of terrestrial origin. Regardless of origin or taxonomic
affiliation, all of the actinomycetes isolated grew luxuriantly in a seawater
based medium and therefore have the potential to not only be metabolically
active in the marine environment but to produce unique metabolites when
provided with specific marine growth factors.
The Associations of Bacteria with Plants and Invertebrates
The distribution of bacteria on living marine surfaces can reveal the extent to
which these organisms are consistently associated with specific plants and
animals. Documenting the specificity of these associations is an integral (but
not in itself conclusive) part of defining bacterial symbioses. In the quest for
new natural products, these associations are particularly significant, because
the potential role of bacterial symbionts in the production of metabolites
previously ascribed to other organisms is open to debate (see section on
bacterial symbioses) (see 37, 39). Without evidence for the consistent associa
tion of bacteria with a given plant or invertebrate, one cannot distinguish
symbionts from transient associates, and the role of symbiotic bacteria in

metabolite production becomes obscured.
Despite increasing documentation of the coexistence between bacteria and
marine invertebrates (see 160), conclusive evidence of symbioses is often
lacking (14). Among the invertebrates frequently collected for natural product
study, sponge-bacterial associations are probably the most thoroughly de
scribed [see review by Wilkinson (194)]. These studies showed that sponge
associated bacteria can be distinct from bacteria in the surrounding seawater
(156, 193, 195). Also, phenotypically similar bacteria were isolated from 9 of
1 0 sponge species collected from two distinct geographical regions (195).
These results suggest that the bacterial communities associated with sponges
may be highly specific. However, this specificity may not apply to all sponges
(e.g. 111).
In the case of coelenterates, another group of organisms typically collected
for chemical study, the mucus of individuals within the Orders Zoanthidae and
Gorgonacea can harbor dense bacterial communities (33, 155, 183). Despite
these observations, little is known about how the communities vary within or
between species. In addition, bacteria associated with algae can be distinct
from bacteria in the surrounding seawater (13, 84, 102, 106), but the specificity
of these associations is not clear. Given the limited number of bacterial asso
ciations described relative to the number of diverse marine organisms available
for study, this subject remains a relatively unexplored area for future research.
Although much remains unknown about bacterial associations with marine
plants and invertebrates, many highly specific associations have been docu
mented. These examples include the associations of bioluminescent Vibrio
fischeri with the light organ of cephalopods (190), dinitrogen-fixing bacteria
with both the gastrointestinal tract of sea urchins (57) and the gland of
Deshayes in shipworms (188), subcuticular bacteria with echinoderms (66,
105), and the spirochete Cristispira sp. with the crystalline style of molluscs
(96). The broad spectrum of these associations suggests that specific bacterial
invertebrate symbioses are common in the sea. Future studies may even show
that the majority of marine organisms have specific bacterial populations

associated with their external surfaces and internal tissues.
A recent observation in our laboratory indicates that unusual and previously
undocumented bacterial associations occur with marine algae. During a study
of bacteria in the coral reef environments of Belize, Central America, we found
that a large percentage of the culturable bacteria associated with algal surfaces
were gram positive. More specifically, 30% of the bacteria associated with the
green alga Halimeda sp. and the brown alga Lobophora variegata were gram
positive (83). This observation contradicts the early belief that 95% of marine
bacteria are gram negative (201). Although gram-positive bacteria have been
reported in greater abundance in sediments (118), the relative abundance of

these bacteria in most marine habitats has not been adequately assessed.
We have included unicellular, gram-positive marine isolates in our studies
of bacterial secondary metabolite production. Of approximately 500 strains
screened thus far, promising insecticidal activity, enzyme production, cyto
toxicity , and at least one novel molecule (178) have been detected. Although
marine gram-positive bacteria do not appear to rival the actinomycetes in their
ability to produce large quantities of structurally complex secondary metabo
lites, additional studies of these bacteria are clearly warranted. As investiga
tions into the distribution of gram-positive and other bacteria on the surfaces
of marine plants and invertebrates continue, previously undocumented asso
ciations may be revealed. These associations may reflect bacterial survival
strategies that include the production of unusual secondary metabolites, as well
as provide important information relating to the bacterial production of meta
bolites previously ascribed to other sources.
The Role of Symbiotic Bacteria in Secondary Metabolite

Production
Historically, the search for marine natural products has centered around chemi
cal studies of marine plants and soft-bodied invertebrates. Considering that
virtually all organisms collected for chemical study include associated micro
organisms, questions about the true biosynthetic origin of molecules isolated
from plants and invertebrates must be addressed. In some cases, circumstantial
evidence leads to the suspicion that metabolites are of bacterial origin. This
evidence can include extremely low (e.g. lO-6% by weight) and often variable
metabolite yields, the isolation of the same metabolite from diverse inverte
brate taxa, consistent microbial associates in the producing organism, and the
similarity of the molecule in question to a previously described microbial
product. Addressing questions of biosynthetic origin are particularly appropri
ate when one considers that in certain marine invertebrates. e.g. sponges,
associated bacteria can account for up to 40% of the cellular volume (194).
Researchers investigating the biosynthetic origins of marine metabolites
have taken several experimental approaches to address this question. The most
common approach has been the isolation and culture of bacteria from the
producing organism followed by chemical analysis of the cultures for the
metabolite of interest. In most cases, these efforts have been unsuccessful,
which is not surprising when one considers the likelihood of obtaining the
producing strain in culture and providing this strain with the culture conditions
necessary for metabolite production. If the bacterial production of a metabolite
cannot be demonstrated in culture, the documentation of a specific bacterial
population associated w ith the producing organism, in an abundance that can
account for the concentration of the metabolite in the host, can provide impor
tant data in support of bacterial b iosynthesis.

Probably the most notable example of the bacterial production of a meta
bolite originally ascribed to another organism involves the potent marine
neurotoxin, tetrototoxin (TTX), best known for the illness and death it can
inflict following the consumption of tainted, raw pufferfish (Fugu spp.). In
vestigations into the origin of TTX (114) revealed that TTX is produced as a
fermentation product of both a Vibrio sp. isolated from the intestines of a
xanthid crab, Atergatis floridus (125), and a Pseudomonas sp. isolated from a
red calcareous alga,Jania sp. (199). More recent studies have shown that TTX
is produced by a variety of taxonomically diverse marine bacteria (31, 1 64,
200), including actinomycetes ( 30). This information has led to the proposal
that bacteria are the sole origin of TTX, which may subsequently accumulate
in invertebrates through the food web ( 31).
In addition to TTX, bacteria have been found to produce at least two other
marine toxins. One example comes from the edib le mollusc Babylonia japon
ica (94), the consumption of which caused at least one outbreak of food
poisoning in Japan. B. japonica toxicity was traced to the presence of neosu
rugatoxin and prosurugatoxin, both of which were isolated from the digestive
gland of the toxic mollusc. Microbiological studies revealed that a coryneform
bacterium,also isolated from the digestive gland of B. japonica, could produce
these toxins. A third example of bacterial toxin production is saxitoxin (STX),
one of the major causative agents in paralytic shellfish poisoning. STX has
been isolated from dinoflagellates, e.g. Protogonyaulax tamarensis (15 8),
which were presumed to be the source of shellfish toxicity. However,shellfish
toxicity is not correlated with the presence of P. tamarensis (129). Recent
studies revealed that a Moraxella sp. isolated from P. tamarensis produces
STX (89-91 ) and may be the u ltimate origin of this metabolite.
In at least two additional studies, the bacterial production of metabolites
previously ascribed to nonbacterial sources has been reported. The first (165)
demonstrated that a Micrococcus sp., isolated from the sponge Tedania ignis,
produced a series of diketopiperazines previously reported from the sponge.
In the second study (35), brominated diphenyl ethers originally isolated from
the sponge Dysidea sp. were obtained as fermentation products from two
strains of Vibrio sp. reported to b e symbionts of Dysidea. Despite such evi
dence of bacterial origin, critical reading of these papers leaves room for
speculation about the true biosynthetic origin of the sponge metabolites (37,
39).
Other attempts to determine the biosynthetic origin of marine metabolites
include the physical separation of bacteria from host tissues,and the subsequent
examination of these cells for the presence of the metabolite in question. The
most successful demonstration of this technique involved the use of flow
cytometry to separate cyanobacterial symbionts (Oscillatoria spongeliae) from

the sponge Dysidea herbacea (181). Through analysis of the purified cyano
bacteria, these authors provided the first evidence for the localization, in
prokaryotic symbiont cells, of a secondary metabolite previously ascribed to
an invertebrate host. Other efforts to determine the cellular location of bacterial
metabolites include the use of molecular probes; however, to the best of our
knowledge these methods have not yet met with success.
Bacterial associations with p lants and animals pose challenging questions
in the field of marine natural products chemistry. Currently, the questions of
greatest interest involve the role of bacteria in the biosy nthesis of metabolites
previously ascribed to other organisms. Part of this interest is purely ecological,
but this subject becomes especially important when the metabolite in question
shows promising biomedical applications. One such case is bryostatin 1, the
anticancer agent isolated from the bryozoan Bugula neritina. In the case of
bryostatin 1, the evidence for bacterial origin includes the low yields (-10-6%
by weight) in which the metabolite is produced and the presence of distinct
populations of bacterial symbionts in the bryozoan tissues. When one considers
that B. neritina is not available from nature in quantities that can effectively
facilitate biomedical development, the possibility of producing bryostatin I by
fermentation becomes a high priority.
Effects of Marine MetaboLites upon the Distribution of Bacteria

Because of the lack of experimental evidence, we can only speculate about
the effects of plant and invertebrate metabolites upon the distribution of bac
teria in the sea. These effects may be expressed in several ways including the
deterrence of u ndesirable bacteria or the attraction of bacteria that represent
favorable epibionts. A lthough laboratory-based assays have indicated that bac
teria can respond to marine metabolites (e.g. 10,20), little ecologically relevant
data are available to suggest these substances have an effect upon surface
colonizing bacteria (see 21 for a review of ecological aspects affecting micro
bial chemotactic behavior).
Early evidence suggesting the involvement of marine metabolites in the
control of epibacterial distributions came from studies of marine algae. In two
separate reports, investigators proposed that antibiotic algal phenols were

responsible for reduced numbers of surface bacteria and fouling organisms on
the marine algae Sargassum natans (162) and Ascophyllum nodosum (25).
Subsequently , a correlation between reduced levels of epibiosis and the pro
duction of antimicrobial substances by marine organisms was presented as
evidence for the ecological role of these metabolites in antifoulIng (e.g. 1,27,
115, 174, 182, 186). Unfortunately,the degree to which antimicrobial activities
translate to in situ effects upon potential surface-colonizing bacteria has not
been adequately demonstrated.
A lthough the in situ microbiological effects of marine natural products

remain largely undocumented, two findings suggest they may influence the
distribution of bacteria on living marine surfaces. One example comes from

the study of two colonial ascidians that possessed remarkably different num
bers of epibiotic bacteria (184). Laboratory settlement assays comparing these
ascidians demonstrated that extracts of the sparsely colonized Cystodytes 10-
batus were highly deterrent to bacterial attachment while extracts of the
densely colonized Polyclinum planum significantly enhanced the attachment
process. These results suggest that the mediation of bacterial attachment by
host metabolites may be an important factor in the control of bacteria upon
living marine surfaces.
A second example demonstrating the effects of secondary metabolites upon
the distribution of microorganisms involves the inhibition of pathogenic fungi
by bacteria associated with the embryos of two marine crustaceans, Palaemon
macrodactylus and Homarus americanus. In separate studies,researchers ob
served that embryos of both P. macrodactylus and H. americanus were covered
by dense assemblages of what appeared to be largely unibacterial populations
(50,51). Bacteria consistently isolated from the embryos of P. macrodactylus
and H. americanus produced the previously described metabolites isatin and
tyrosol, respectively, both of which show antifungal effects toward the crus
tacean pathogen,Lagenidium callinectes. Further experiments with P. macro
dactylus revealed that bacteria-free embryos rapidly succumb to fungal
infection whereas similar embryos re-inoculated with bacteria survived. These
results led to the conclusion that symbiotic bacteria chemically defend P.
macrodactylus embryos against fungal infection. Bacterial associations of this
nature add a new level of complexity to the mechanisms by which secondary
metabolites influence the distributions and specific associations of marine
microorganisms.
The Adaptations of Bacteria to Diverse Marine Habitats

Their metabolic capacity allows prokaryotes to survive virtually across the
spectrum of environmental conditions found on this planet. Assuming that
metabolic and biosynthetic di versity are correlated, the adaptations of bacteria
to diverse marine habitats provide seemingly limitless evolutionary opportu

nities for the production of unique secondary metabolites. Although bacteria
adapted to diverse marine environments have apparently received little atten
tion in the search for novel secondary metabolites, an important industrial
application has corne from the study of one such group, i.e. the thermophilic
archaea (Archaebacteria) associated with deep-sea hydrothermal vents (SO).
These bacteria produce thermostable DNA polymerase enzymes, which have
been isolated, cloned, and marketed for biotechnological applications (7S).
Other evidence of bacterial adaptations to extreme marine environments in
cludes the recognition that a gene (ompH) from a gram-negative bacterium

isolated from the depths of the Sulu Sea is expressed only at increased hydro
static pressure (9). This discovery suggests the existence of a genetic program
that couples pressure sensing to protein synthesis.
Bacterial Culturability
Maintaining bacteria in culture is requisite to the effective study of secondary
metabolite production, because adequate quantities of b acterial biomass can
seldom be obtained from samples collected in the field. Unfortunately, the
majority of bacteria observed in certain marine samples do not form colonies
on the nutrient-rich agar traditionally used for the isolation of marine bacteria
(81,92). In fact, microscopic counts performed directly on seawater samples,
usually accomplished with florochrome stains (e.g. 64, 146), typically yield
bacterial counts three orders of magnitude higher than those obtained with
plating techniques (e.g. 163). These results have led to the generally accepted
conclusion that the majority of marine bacteria are unculturable,a conclusion
that raises serious concerns for researchers interested in maximizing the num
ber of diverse isolates screened for unique secondary metabolites.
Several topics deserve closer scrutiny in the discussion of bacterial cul
turability. For one, most studies have focused upon the b acteria in seawater
samples collected far from shore. This may be important considering that the
organic content of open-ocean seawater is generally low (116),and that seawa
ter bacteria have evolved survival strategies (154), including responses to
starvation (97, 127), that may dramatically reduce their ability to form colonies
on nutrient-rich agar (e. g. Difco 2216). Furthermore,the discrepancy between
viable and direct counts, for some seawater samples, may result from low
plating efficiencies, i.e. only a few viable individuals in a population are
capable of forming colonies on agar media (147).
It should also be noted that the percentage of culturable b acteria calculated
for marine samples can be higher than the 0.1% typically reported for seawater.
For example, 3.4 to 11% of the bacteria observed in association with the
sclerosponge Ceraloporella nicholsoni were culturable (156). If living marine
surface s provide nutrient-rich environments, a relatively large percentage of
the bacteria associated with these surfaces may form colonies when inoculated
onto nutrient-rich growth media. To the best of our knowledge, the culturablil
ity of bacteria associated with nutrient-rich marine habitats has not been ad
dressed in detail. One habitat that would be ideal for such studies is the marine
invertebrate digestive system, which can harbor extensive bacterial communi
ties (61).
Another concept that must be kept in mind in discussions of bacterial
culturability is the inherent selectivity of all isolation methods. Because only
a limited group of bacteria will grow when any one isolation method is used,
it is not surprising that large numbers of bacteria appear to be unculturable.

To determine the percentage of culturable bacteria in any community, one
would have to attempt multiple isolation methods and determine the sum of
all unique strains resulting from these methods. This is not feasible; hence the
percentage of culturable bacteria reported for any one sample must be consid
ered in the context of the isolation methods used.
We can also ask questions about the composition of the nonculturable
component of the bacterial community. Do these bacteria represent noncultur
able strains of previously described species, or do they represent new species
or genera? Recent analyses of small subunit ( 1 6S ) bacterial rRNA sequences
have shed light on this subject. For example, it has been shown that bacte
rioplankton communities consist of broadly diverse bacterial assemblages
within which undescribed phylogenetic groups are common (43). As we be
come aware of novel bacterial populations, focused efforts can be made to
assess the culturability of these bacteria.
Apparently, for taxonomically diverse marine bacteria to be effectively
incorporated into the search for new microbial products, new culture methods
need to be developed, particularly methods that take into account the environ
mental parameters associated with the habitats sampled. These methods may
include the use of marine-derived nutrients, which have been shown to induce
the production of novel metabolites in at least one report ( 1 36). In this study,
the addition of a pulverized brown alga, kobu cha (Laminaria sp.), to the
growth medium induced the production of a new antibiotic, SS-228 Y. The
development of marine-derived nutrients may improve our ability to culture
marine bacteria and present additional opportunities for the discovery of novel
metabolites.
The Distinction Between Marine and Terrestrial Bacteria

Establishing the existence of specific marine bacteria was fundamental to the
development of marine microbiology as a scientific discipline. Early studies
led to the proposal that marinc bactcria can bc distinguishcd from terrestrial
strains based upon the requirement of seawater for growth (202). Subsequent
studies documented a readily detectable sodium requirement for growth, which
is rarely observed for terrestrial strains (110). Although marine bacteria lack
an official definition, this demonstration of a specific sodium requirement for
growth has become widely accepted as a method to distinguish marine from
most terrestrial bacteria [see reviews by MacLeod (l08, 109)].
More recently, the physiological role of sodium was investigated in greater
detaiL For many marine bacteria,the transport of various solutes into the cell
is sodium dependent (11, 175, 196). The energy required for this transport i s
gen erated by a respiration-dependent sodium pump (175). This pump extrudes
sodium ions,generating a transmembrane sodium gradient with potential en
ergy that can be utilized to perform various cellular functions including the
generation of ATP. Oh et al (130) have shown that the sodium requirement in

marine bacteria is correlated with the possession of respiration-dependent
sodium efflux,and these authors have suggested that electron transport-driven
sodium efflux can b e regarded as a criterion for the definition of marine
bacteria.
Although the physiological distinctions between marine and terrestrial bac
teria are clearly important, the rationale for restricting marine bacteria to only
those strains that meet a predefined set of requirements is questionable. For
example, in studi es conducted off the coast of Belize, we observed that more
than 80% of the gram-positive bacteria isolated from a variety of sources
required seawater for growth (83). In addition, all seawater-requiring strains
tested had a demonstrable requirement of sodium for growth. As a result, these
bacteria can be defined as marine based upon traditional proposals (110, 202).
Also based on these proposals,bacteria that did not require seawater or sodium
for growth,but w ere capabl e of growth in seawater-based medium,could be
labeled as halotolerant terrestrial strains. But,should w e reach this conclusion
a priori, and as a result eliminate non-seawater- and/or non-sodium-requiring
bacteria from discussions of marine bacteria? To take this question one step
further, we can envision a number of marine niches, e.g. those inhabited by
intracellular symbionts, in which bacteria may never encounter seawater. If
these bacteria do not have a demonstrable sodium requirement, aren't they
nonethel ess marine? We believe that by restricting marine bacteria to include
only those strains that meet a predefined set of requirements we run the risk
of overlooking certain groups that may be equally well adapted to survive on
land or in the sea, as w ell as indigenous marine bacteria that simply do not
meet the set of requirements chosen.
Furthermore,the requirement of seawater for the growth of marine bacteria
appears to vary between taxa. In the case of gram-positive bacteria, none of
the cocci tested in at least four separate studies required seawater for growth
(83, 93, 100, 197). Also,as currently described the genus Planococcus, which
is recognized as a marine genus, does not require seawater for growth (23, 88,
128). MacLeod (109) makes an interesting observation regarding the gram-
positive coccus Gaffkya homari (now Aerococcus viridans), the pathogenic

bacterium that causes septicemia in lobsters. Citing unpublished data, he states
that G. homari does not require seawater for growth and that at least among
the gram-positive species in the sea, some representatives of the indigenous
flora require neither seawater nor sodium for growth. With the exception of
the moderately halophilic cocci, e.g.Marinococcus spp., Salinococcus spp.,
Sporosarcina halophila, and Micrococcus halobius (23, 60, 1 1 3), which grow
optimally in 20% salt solutions and are typically isolated from solar salterns,
salted food products, and other hypersaline environments, there appears to be
a trend in which gram-positive cocci isolated from the sea do not require
seawater or sodium for growth. This subject certainly warrants further inves
tigation.
In our opinion, to make a distinction between marine and terrestrial bacteria
based upon sodium requirements is not as important for natural product dis
covery as the demonstration that bacteria isolated from the sea perform unique
biosyntheses. The best example of unique marine biosyntheses may well be
the bacterial production of halogenated antibiotics (e.g. 2, 1 8, 1 40). If the
production of these and other metabolites that possess features seldom encoun
tered outside the realm of the ocean is restricted to a specific set of marine
conditions, then chemical studies of bacteria grown in these conditions are
warranted.
BIOLOGICAL ACTIVITIES AND NOVEL ORGANIC

SUBSTANCES
The ability of marine bacteria to produce antibacterial substances was docu

mented more than 45 years ago ( 1 53). Nearly 20 years later, Burkholder and
coworkers ( 1 8) purified the first antibiotic from a marine bacterium, and later
that same year, Lovell ( 107) reported the structure of this substance, a highly
brominated pyrrole. In the years that followed these early discoveries, the
number of novel organic molecules reported from marine bacteria has grown
to include nearly 30 substances (Table 1 ). These substances represent diverse
classes of chemical structures and provide evidence for the untapped biosyn
thetic potential of marine bacteria. Although the total number of unique mole
cules reported to date remains low, approximately one half of these were
described after 1 990, reflecting a growing interest in this area of research.
The biological activities reported for the molecules described in Table 1
include not only antibiotic, but also anticancer and antiviral activities. To some
extent, these activities are a reflection of bioassay accessibility and the scien
tific objectives of the research group responsible for the discovery. Although
the rapid screening of bacterial extracts is essential to biomedically relevant
discovery (because of low hit rates) and to effective bioassay-guided chemical

isolation, these bioassays can become burdensome for small research groups.
Maximizing the potential for drug discovcry through in-house testing is also
difficult for small research groups considering the many new and sophisticated
target-directed screens developed in recent years. For these reasons, industrial
and academic collaborations offer an effectiv e avenue by which to obtain
broad-spectrum screening of bacterial metabolites. As efforts to screen marine
microbial products increase,additional biomedically and industrially important
activities will c ertainly be discovered.
We mentioned that novel secondary metabolites have been isolated from a

limited taxonomic group of marine bacteria. This may be misleading as othcr
genera, including Moraxella (52) and Alcaligenes ( 1 6) produce biologically

active substances that have not been fully characterized. In addition,an anti
tumor polysaccharide has b een isolated from a Flavobacterium sp. ( 1 80), and
antibiotic activity was associated with a purple photosynthetic bacterium,
Chromatium purpuratum ( 1 7). However, to the best of our knowledge many
taxa have not been studied in detail for the production of biologically active
secondary metabolites.
One genus that is conspicuously absent from the marine natural products
literature is Bacillus, even though isolates are readily obtained from marine
sediments ( 1 6 1 ) and unusual secondary metabolites have been reported from
terrestrial strains ( 1 2). Other than the recent isolation in our laboratory of a
previously undescribed cyclic depsi-peptide ( 1 78), to the best of our knowl
edge,only one other substance-an enzyme-has been reported from a marine
Bacillus sp. ( 1 33). The archaea were also absent from the marine natural
products literature prior to the recent isolation of new cyclic polysulfides from
strains of anaerobically cultured Thermococcus spp. ( 1 52). These thermophilic,
sulfur-metabolizing archaea were collected near marine hydrothermal systems
and the substances produced demonstrated both antifungal and anthelmintic
acti vities.
In addition to the cases already mentioned,the history of biological activities
reported for marine bacteria is long. For example, after the survey of antibiotic
activity by Rosenfeld & Zobell ( 153) came demonstrations that marine bacteria
possess antiviral (52, 1 76), anti-algal (62), anticancer ( 1 80), and antimicrobial
activities (6, 7, 1 5-17, 32, 44-48, 56, 95, 103, 104, 1 1 9). Investigators have
also demonstrated that marine bacteria produce bacteriocins (68) and enzyme
inhibitors (70-75, 1 7 1). In some cases, these activities are due to proteins or
other large molecules for which structural elucidation cannot be performed
using standard spectral techniques. In other cases, attempts to isolate and
characterize the active principles were never made. The extent of these activi
ties, however, supports further investigation of secondary metabolites from
marine bacteria.
CONCLUDING REMARKS
Today, marine bacteria are being recognized as an important resource for

microbial products. The growing interest in this resource is adequately dem
onstrated by the number of novel metabolites recently reported from marine
bacteria and the number of research laboratories throughout the world now
working in this field. For the chemical potential of marine bacteria to be
effectively assessed, however, continued basic research in marine microbiol
ogy is essential. This research· is needed because we do not yet have a clear
understanding of the types of marine bacteria available for chemical study, the
niches they occupy, and the mechanisms by which they are adapted to the
marine environment.
The focus of our research in marine microbiology is to isolate novel secon
dary metabolites from marine bacteria and to ask ecological questions about
both the producing strains and the roles of the metabolites produced. In the
course of this work, it seems appropriate to consider our results in terms of
traditional concepts in marine microbiology. For example, should we distin
guish marine from terrestrial bacteria based upon sodium ion requirements, or
for that matter, any specific physiological criteria? Although the elucidation
of specific ion requirements and the function of these ions in bacterial growth
is clearly important from a physiological perspective, these adaptations may
prove to be less important, in an ecological context, than the demonstration
that bacteria are metabolically active in the marine environment. Certainly in
terms of natural product discovery, a marine isolate's ability to produce sec
ondary metabolites only when provided with conditions specific to the marine
environment is a primary consideration.
Another general concept in marine microbiology that can be addressed in
greater detail, especially in light of available modem techniques, is the number
and activity of gram-positive bacteria in the sea. The relative numbers of
gram-positive bacteria in some marine habitats may be greater than previously
believed. For example, a recent study demonstrated that 9 of 6 1 clones, ob
tained from seawater samples and examined by small subunit ( 16S) rRNA
sequence analysis, clustered within a group of gram-positive bacteria (43).
Although this study was not intended to be quantitative, it suggests that gram
positive bacteria may represent a numerically significant component of the
total viable as well as the culturable (83) bacterial community. Clearly, addi
tional descriptions of gram-positive bacteria are needed to better clarify their
numbers and activities in the sea.
To date, only the most preliminary investigations have been made in the
search for novel metabolites from marine bacteria. From an academic perspec
tive, many ecological questions fundamental to this field remain unanswered.
These q uestions include the effects of plant and invertebrate metabolites up on
SECONDARY METABOLITES FROM MARINE B ACTERIA 577
the distribution of marine bacteria and the roles of these metabolites in chemi
cal defense against microbial pathogens. Other important questions address
the role of symbiotic bacteria in the production of metabolites previously
ascribed to nonbacterial sources. Without continued ecological studies of ma
rine bacteria, their potential for the production of novel secondary metabolites
may never be fully realized.
ACKNOWLEDGMENTS
We thank Professors Ralph Mitchell and Farooq Azam for their critical evalu
ation of this manuscript. In addition, we thank Carole Bewley for helpful

editorial comments.
Any Annual Review chapter, as well as any article cited in an Annual Review chapter,
may be purchased from the Annual Reviews Preprints and Reprints service.
1 .800.347.8007; 415.259.5017; email: arpr@eiass.org
Literatur e Cited
1. AI-Ogily SM, Knight Jones EW. 1977.

- 10. Bell W, Mitchell R. 1972. Chemotactic
Antifouling role of antibiotics produced and growth responses of marine bacteria
by marine algae and bryozoans. Nature to algal extracellular products. BioI.
265:728-29 Bull. 143:265-77
2. Andersen RJ, Wolfe MS, Faulkner DJ. 11. Berthelet M, MacLeod RA. 1 99 1 . The
1974. Autotoxic antibiotic production role of Na' in membrane transport and
by a marine Chromobacterium. Mar. respiration in the marine bacterium
Bioi. 27:281-85 Deleya aesta 1 34 . Can. J. Microbiol.
3. Attaway DH, Zaborsky OR, eds. 1993. 37:433-39
Marine Biotecnology, Vol. 1, Pharma 1 2. Betina V. 1 9 8 3 . The Chemistry and
ceutical and Bioactive Natural Prod· Biology of Antibiotics, Vol. 5. New
ucts. New York: Plenum. 500 pp. York: Elsevier. 591 pp.
4. Austin B. 1988. Marine Microbiology. 13. Bolinches J, Lemos ML, Barja JL. 1988.
New York: Cambridge Univ. Press Population dynamics of heterotrophic
5. Austin B . 1989. Novel pharmaceutical bacterial commuI1ities associated with
compounds from marine bacteria. J. Fucus vesiculosus aI1d Ulva rigida in
Appl. Bacteriol. 67:461-70 an estuary. Microb. Ecol. 15:345-57
6. 8 aam RB, Gandhi NM, Freitas YM. 14. Bonar, DB, Weiner RM, Colwell RR.
1966. Antibiotic activity of marine mi 1986. Microbial-invertebrate i nterac
croorganisms: the antibacteri al spec tions and potential for biotechnology .

trum. Helgol. Wiss. Meeresunters. 1 3 : Microb. Ecol. 1 2 : 101-10
1 88-91 15. Buck, JD, Ahern DG, Roth FJ Jr, Mey
7. Ballester M , Ballester JM, Belaich JP. ers SP. 1963. Inhibition of yeasts by a
1977. Isolation and characterization of marine bacterium. J. Bacteriol. 85:
a high molecular weight antibiotic pro· 1 1 32-35
dnced by a marine bacterinm. Microb. 16. Buck 10, Meyers SP, Kamp KM. 1 962.
Ecol. 3 :289-303 Marine bacteria with antiyeast activity.
8. Barcina I, Iriberri J, Egea L. 1987. Enu Science 2 1 : 1 339-40
meration, isolatioI1 and some physi 17. Burgess JG, Mi yashi ta H, Sudo H, Mat
ological properties of actinomycetes sunaga T. 1 99 1 . Antibiotic production
from seawater and sediments. Syst. Appl. by the marine photosynthetic bacterium
Microbiol. 10: 85-91 Chromatium purpuratum NKPB
9. Bartlett D, Wright M, Yayanos AA, 03 1 704: localization of activity to the
Silverman M. 1989. Isolation of a gene chromatophores . FEMS Microbiol. Lett.
regulated by hydrostatic pressure in a 84:30 1-6
deep sea bacterium. Nature 342:572-74
- 18. Burkholder PR, Pfister RM, Leitz FP.
1966. Production of a pyrrole antibiotic Identification of deep-sea-sediment bac

by a marine bacterium. Appl. Microbiol. teria which produce tetrodotoxin. Appl.
14:649-53 Environ. Microbial. 56: 1 1 62-63
19. Bushell ME. Grafe U. eds. 1989. Bioac 32. Doggett RG. 1968. New anti Pseudo-
tive Metabolites from Microorganisms, monas agent isolated from a marine

Progress in Industrial Microbiology Se Vibrio. J. Bacteriol. 95 : 1972-73
ries, Vol. 27. AmsterdamlNew York: 33. Ducklow HW. Mitchell R. 1979. Bac
Elsevier. 4 19 pp. terial populations and adaptations in the
20. Chet I. Asketh p. Mitchell R. 1975. mucus layers on living corals. Limnol.
Repulsion of bacteria from marine sur Oceanogr. 24:71 5-25
faces. Appl. Microbiol. 30: 1 043-45 34. EI-Nakeeb MA, Lechevalier HA. 1962.
21. Chet I, Mitchell R. 1976. Ecological Selective isolation of aerobic actinomy
aspects of microbial chemotactic be cetes. Appl. Microbiol. 1 1 :75-77
havior. Annu. Rev. Microbiol. 30:221- 35. Elyakov GB, Kuznetsova T. Mikhailov
39 VV. Mal'tsev II, Voinov VG, Fedoreyev

22. Chisholm SW, Olson RJ, Zettler ER, SA. 199 1 . Brominated diphenyl ethers
Goericke R, Waterbury ]B, Welsch from a marine bacterium associated with

meyer NA. 1988. A novel free-living the sponge Dysidea sp. ExperieTltia 47:
prochlorophyte abundant in the ocean 632-33
euphotic zone. Nature 334:340-43 36. Faulkner DJ. 1993. Marine natural prod
23. Claus D, Fritze D, Kocur M. 1 992. ucts. Nat. Prod. Res. 10:497-539
Genera related to the genus Bacillus 37. Faulkner JD, He H, Unson MD, Bewley
Sporolactobacillus, SporoscarciTlil, Plmw CA. 1993. New metabolites from marine
coccus, Filibacter, and Caryophanon. sponges: are symbionts important?
In The Procaryotes. a Handbook on the Cazz. Chirn. Ital. 123:301-7
Biology of Bacteria: Ecophysiology, 38. Fautin DG, ed. 1988. Biomedical Im
Isolulion, Identification, Applications, portance of Marine Organisms. Mem.
ed. A Balows, HG Truper, M Dworkin, Calif. Acad. Sci. 1 3 : 1 1 59
-
W Harder, K Schleifer, 2: 1 769-84. New 39. Fenical W. 1993. Chemical studies of

York: Springer-Verlag. 2nd ed. marine bacteria. Chern. Rev. 93: 1 673-83
24. Corpe W A. 1 970. Attachment of marine 40. Fenical W, Jensen PR. 1993. Marine
bacteria to solid surfaces. In Adhesions microorganisms: a new biomedical re
in Biological Systems, ed. RS Manly, source. See Ref. 3. pp. 4 19-75
pp. 73-87. New York: Academic 41. Fletcher M, Marshall KC. 1982. Are
25. Cundell AM, Sleeter TD, Mitchell R. solid surfaces of ecological significance
1977. Microbial populations associated to aquatic bacteria? Adv. Microbial.
with the surface of the brown alga As Eco!' 6 : 1 99-236
cophyllum nodosum. Microbial Eco/. 42. Fuhrman JA. McCallum K. Davis AA.
4:81-9 1 1992. Novel major archaebacterial
26. Davidson B S , Schumacher RW. 1993. group from marine plankton. Nature
Isolation and synthesis of caprolactins 356: 148-49
A and B, new caprolactams from a 43. Fuhrman JA, McCallum K, Davis AA.
marine bacterium. Tetrahedron 49: 1993. Phylogenetic diversity of subsur
6569-74 face marine microbial communities
27. Davis AR, Targett NM, McConnell OJ, from the Atlantic and Pacific Oceans.
Young CM. 1989. Epibiosis of marine Appl. Environ. Microbio!' 59:1 294-
algae and benthic invertebrates: natural 1302
products chemistry and other mecha 44. Fusetani N, Ejima D, Matsunaga S,
nisms inhibiting settlement and over Hashimoto K, Itagaki K, et al. 1987.
growth. See Ref. 158a, pp. 85-1 1 4 3-Amino-3-deoxY-D-glucose: an antibi
28. DeLong EF. 1992. Archaea in coastal otic produced by a deep-sea bacterium.
marine environments. Proc. Natl. Acad. Experientia 43:464-65
Sci. USA 89:5685-89 45. Gauthier MJ. 1976. Alteromonas rubra
29. Demain AL, Solomon NA, eds. 1986. sp. nov., a new marine antibiotic-pro
Manual of Industrial Microbiology and ducing bacterium. Int. J. Syst. Bacteriol.
Biotechnology. Washington, DC: Am. 26:459-66
Soc. Microbiol. 466 pp. 46. Gauthier MJ. 1 976. Modification of bac
30. Do HK, Kogure K, Imada C, Noguchi terial respiration by a polyanionic anti
T. Ohwada K. Simidu U. 1 99 1 . Tetro biotic produced by a marine Allero
dotoxin , production of actinomycetes monas. Antimicrob. Agents Chemother.
isolated 'rrbm marine sediment. 1. Appl. 9:361-66
Bacteriol. 70:464-68 47. Gauthier MJ. Breittmayer VA. 1979. A
31. Do HK, Kogure K, Simidu U . [ 990. new antibiotic-producing bacterium
from seawater: Alteromonas aurantia ner) comb. nov. J. Gen. Appl. Microbiol.
sp. nov. Int. J. Syst. Bacteriol. 29:366- 30:449-59
72 61. Harris 1M. 1993. The presence, nature,
48. Gauthier MJ, Flatau GN. 1976. Antibac and role of gut microflora in aquatic
terial activity of marine violet-pig invertebrates: a synthesis. Microb. Ecol.
mented Alteromonas with special 25: 195-231
reference to the production of bromi 62. Hayashida S, Tanaka S, Teramoto Y,
nated compounds. Can. J. Microbial. Nanri N, Yoshino S, Furukawa K. 1 99 1 .
22: 1 6 1 2-19 Isolation o f anti-algal Pseudomonas
49. Gerber NN. 1983. Cyc1oprodigiosin stutzeri strains and their lethal activity
from Beneckea gazogenes. Tetrahedron for Chattonella antiqua. Agric. BioI.
Letl. 24:2797-98 Chern. 3 :787 90
-
50. Gil-Turnes MS, Fenical W. 1 992. Em 63. Helmke E, Weyland H. 1984. Rhodo
bryos of Homarus americanus are pro coccus marinonascens sp. nov., an act
tected by epibiotic bacteria. Bioi. Bull. inomycete from the sea. Int. J. Syst.
1 82: 1 05-8 Bacterial. 34:127-38
51. Gil-Turnes MS, Hay ME, Fenical W. 64. Hobbie JE, Daley RJ, Jasper S. 1977.
1989. Symbiotic marine bacteria chemi Use of nucleopore filters for counting
cally defend crustacean embryos from bacteria by flourescence microscopy.
a pathogenic fungus. Science 246: 1 17- Appl. Environ. Microbiol. 33 : 1225-28
18 65. Holland GS, Jamieson DD, Reichelt JL,
52. Girones R , Jofre JT, Bosch A. 1989. Viset G, Wells RJ. 1984. Three aromatic
Isolation of marine bacteria with antivi acids from a marine bacterium. Chem.
ral properties. Can. J. Microbial. Ind. 23 : 850-5 1
35: 1 0 1 5-21 66. Holland ND, Nealson KH. 1978. The
53. Goodfellow M . Haynes lA. 1 984. Act fine structure of the echinoderm cuticle
inomycetes in marine sediments. In Bio and the subcuticular bacteria of echino
logical. Biochemical. and Biomedical derms. Acta Zool. 59: 169-85
Aspects of Actinomycetes. ed. L Ortiz 67. Hotta K, Yoshida M. Hamada M,
Ortiz, LF Bojalil, V Yakoleff, pp. 453- Okami Y. 1980. Studies on new ami
72. Orlando: Academic noglycoside antibiotics, istamycins,
54. Goodfellow M, Williams E. 1 986. New from an actinomycete isolated from a
strategies for selective isolation of in marine environment. J. Antibiot. 33:
dustrially important bacteria. Biotech 1 5 1 5-20
nol. Genet. Eng. Rev. 4:2 1 3-62 68. Hoyt PR, Sizemore RK. 1982. Competi
55. Goodfellow M, Williams ST . 1983. tive dominance by a bacteriocin-produc
Ecology of actinomycetes. Annu. Rev. ing Vibro harveyi strain. Appl. Environ.
Microbial. 37: 1 89-21 6 Microbiol. 44:653-58
56. Grein A. Meyers S P . 1958. Growth 69. Hsu SC, Lockwood JL. 1975. Powdered
characteristics and antibiotic produc chitin as a selecti ve medium for enu
tion of actinomycetes isolated from meration of actinomycetes in water and
littoral sediments and materials sus soil. Appl. Microbial. 29:422-26
pended in seawater. J. Bacteriol. 76: 70. Imada C, Maeda M, Hara S, Taga N,
457-63 Simidu U. 1986. Purification and char
57. Guerinot ML, Patriquin DG. 1 98 1 . The acterization of subtili si n inhibitors 'Ma
association of N,-fixing bacteria with rinostatin' produced by marine Altero
sea urchins. Mar. Bioi. 62: 197-207 monas sp. J. Appl. Bacteriol. 60: 469-76
58. Gustafson K, Roman M, Fenical W. 71. Imada C, Maeda M, Taga N. 1985.
1989. The macrolactins, a novel class Purification and characterization of the
of antiviral and cytotoxic macrolides protease inhibitor 'monastatin' from a
from a deep-sea marine bacterium. J. marine Alteromonas sp. with reference
Am. Chern. Soc. 1 1 1 :75 1 9-24 to inhibition of the protease produced
59. Hamato N, Takano R, Kamei-Hayashi by a bacterium pathogenic to fish. Can.
K, Imada C, Hara, S. 1992. Leupeptins J. Microbiol. 3 1 : 1 089-94
produced by the marine Altermonas sp. 72. Imada C, Simidu U. 1988. Isolation and
B-IO-3 1 . Riosci. Biotech. Biochem. characterization of an a-amylase inhibi
56: 1 3 1 6-18 tor producing actinomycete from marine
60. Hao MV, Kocur M, Komagata K . 1984. environment. Nippon Suisan Gakkaishi
Marinococcus gen . nov.� a new genus 54: 1 839--45
for motile cocci with meso-diaminopi 73. lmada C, Simidu U. 1992. Culture con
metic acid in the cell wall; and Mari ditions for an a-amylase inhibitor-pro
IlOCOCCuc albus sp. nov. and Murillo ducing marine actinomycete and pro
coccus haluphilus (Novitsky and Kush- duction of the inhibitor 'amylostrep-
tin.' Nippon Suisan Gakkaishi 58: 2 1 69- logical properties. l. Antibiot. 40: 1 664-
74 70
74. Imada C, Simidu U, Taga N. 1985. 86. Kitahara T, Naganawa H, Okazaki T,
Isolation and characterization of marine Okami Y, Umezawa H. 1975. The struc
bacteria producing alkaline protease in ture of SS-228Y, an antibiotic from
hibitor. Bull. lpn. Soc. Sci. Fish. Chainia sp. l. Antibiot. 28:280-85
5 1 :799-803 87. Kobayashi J, Ishibashi M. 1 993. Bioac
75. Imada C, Taga N, Maeda M. 1985. tive metabolites of symbiotic marine
Cultivation conditions for subtilisin in microorganisms. Chem. Rev. 93: 1 753-
hibitor-producing bacterium and general 69
properties of the inhibitor ' Mari 88. Kocur M , Pacova Z, Hodgkiss W,
nostatin.' Bull. lpn. Soc. Sci. Fish. 5 1 : Martinec T. 1970. The taxonomic status
805- 1 0 of the genus Planococcus Migula 1 894.
76. Imamura N, Nishijima M, Adachi K, Int. l. Syst. Bacteriol. 20 :24 1 -4 8
Sano H. 1993. Novel antimycin antibi 89. Kodama M, Ogata T, Sakamoto S, Sato
otics, urauchimycins A and B, produced S, Honda T, Miwatani T. 1 990. Produc
by manne actinomycete. l. Antibiot. tion of paralytic shellfish toxins by a

46:241-46 bacterium Moraxella sp. isolated from
77. Ireland CM, Copp BR, Foster MP, Protogonyaulax tamarensis. Toxicon
McDonald LA, Radisky DC, Swersey 28:707-1 4
Je. 1993. Biomedical potential of ma 90. Kodama M , Ogata T , Sato S . 1988.
rine natural products. See Ref. 3, pp. Bacterial production of saxitoxin. Agric.
1-43 BioI. Chern. 52: 1 075-77
78. Jack WE, Kucera RB, Kong H . 1992. 91. Kodama M, Ogata T, Sato S, Sakamota
Biochemical characterization of VentR S. 1 990. Possible association of marine
and Deep VentR DNA polymerases. New bacteria with paralytic shellfish toxicity
Engl. Biolabs Transcr. 4:4-5 in biv alves . Mar. Ecol. Prog. Ser.
79. lalal MAF. Hossain MB. van def Helm 6 1 :203-6
D, Sanders-Loehr J , Actis LA, Crosa 92. Kogure K, Simidu U, Taga N. 1979. A
J H . 1989. Structure of anguibactin, a tentative direct microscopic method for
unique plasmid-related bacterial sidero counting living marine bacteria. Can. l.
phore from the fish pathogen Vibrio Microbiol. 25:415-20
anguillarum. l. Am. Chem. Soc. I l l : 93. Kong MK, Chan K. 1979. A study on
292-96 the bacterial flora isolated from marine
80. Jannasch HW. 1 992. Deep sea hy algae. Bot. Mar. 22:83-97
drothermal vents: underwater oases. 94. Kosuge T, Tsuji K, Harai K, Fukuyama
New Engl. Biolabs Transcr. 4 : 1-3 T. 1985. First evidence of toxin produc
81. Jannasch HW, Jones GE. 1959. Bacte tion by bacteria in a marine organism.
rial populations in seawater as deter Chem. Pharm. Bull. 33:3059-61
mined by different methods of 95. Krasil' nikova EN. 1 96 1 . Antibiotic
enumeration. Limnol. Oceanogr. 4: 1 28- properties of microorganisms isolated
39 from various depths of world oceans.
82. Jensen PR, Dwight R, Fenical W. 1 99 1 . Mikrobiologiya 30:65 1-57
Distribution o f actinomycetes i n near 96. Kuhn DA. 1 9 8 1 . The genus Cristispira.
shore tropical marine sediments. Appl. In The Prokaryotes, ed. MP Starr, H
Environ. Microbiol. 57: 1 102-8 Stolp, HG Trtiper, A Balows, HG
83. Jensen PR, Fenical W. 1994. The rela Schlegel, I :555-63. Berlin: Springer
tive abundance and seawater require Verlag. 1 102 pp.
ments of Gram-positive bacteria in 97. Kurath G, Morita Y. 1983. Starvation
near-shore tropical marine samples. Mi survival physiological studies of a ma
crob. Ecol. In press rine Pseudomonas sp. Appl. Environ.
84. Johnson CR, Muir DG, Reysenbach AL. Microbiol. 45: 1 206- 1 1
1 99 1 . Characteristic bacteria associated 98. KUster E, Neumeier W. 1 98 1 . Halotol
with surfaces of coralline algae: a hy erance in some streptomycetes produc
pothesis for bacterial induction of ma ing tetracyclines. In Actinomycetes, ed.
rine invertebrate larvae. Mar. Ecol. KP Schaal, G Pulverer. Zentralbl. Bak
Prog. Ser. 74:281-94 teriol. Mikrobiol. Hyg. Abt. 1. 1 1 : 3 1 5-
85 . Kameyama T, Takahashi A, Kurasawa 19 (SuppL)
S, Ishizuka M, Okami Y, et al. 1987. 99. KUster E, Williams ST. 1964. Selection
Bisucaberin, a new siderophore sensi of media for isolation of streptomycetes.
tizing tumor cells to macrophage-medi Nature 202:928-29
ated cytolysis. I. Taxonomy of the 100. Kusuda R, Kawai K, Salati F, Banner
producing organism, isolation and bio- CR, Fryer JL. 1 99 1 . Enterococcus seri-
SECONDARY METABOLITES FROM MARINE BACTERIA 58 1
olicida sp. nov., a fish pathogen. Int. J. bripes. Bull. Jpn. Sac. Sci. Fish. 47:535-
Syst. Bacterial. 4 1 :406-9 37
101 . Lappin-Scott HM, Costerton JW. 1989. 1 15 . McCaffrey EJ, Endean R. 1985. Antimi
Bacterial biofilms and surface fouling. crobial activity of tropical and subtropi
Biofouling 1:3 2�2 cal sponges. Mar. BioI. 89: 1 -8
102. Laycock RA. 1 974. The detrital food 1 16. Menzel DW, Ryther J. 1 %8. Distribu
chain based on seaweeds. I. Bacteria tion and cycling of organic matter in
associated with the surface of Laminaria the oceans. In Symposium on Organic
fronds. Mar. Bioi. 25:223-31 Matter in Natural Waters, ed. DH Hood,
103. Lemos ML, Dopazo CP, Toranzo AE, 1 :3 1-54. College, AK: Univ. Alaska
Barja JL. 1 99 1 . Competative dominance 1 17 . Mitchell R, Kirchman D. 1984. The
of antibiotic-producing marine bacteria microbial ecology of marine surfaces.
in mixed cultures. J. Appl. Bacterial. I n Marine Biodeteriaration: An Inter
7 1 :228-32 disciplinary Study, ed. JD Costlow, RC
104. Lemos ML, Toranw AE, Barja JL. Tipper, pp. 49-58. London: E & FN
1985. Antibiotic activity of epiphytic Spon. 384 pp.
bacteria isolated from intertidal sea 1 1 8. Moriarty DJW, Hayward AC. 1982. Ul
weeds. Microb. Ecol. 1 1 : 149-63 trastructure of bacteria and the propor
105. Lesser MP, Blakemore RP. 1 990. De tion of Gram-negative bacteria in marine
scription of a novel symbiotic bacterium sediments. Microb. Ecal. 8: 1-14
from the brittle star, Amphipholis 1 19. Nair S, Simidu U. 1987. Distribution
squamata. Appl. Environ. Microbial. and significance of heterotrophic marine
56:2436-40 bacteria with antibacterial activity. Appl.
106. Lewis TE, Garland CD, McMeekin TA. Environ. Microbial. 53:2957-62
1985. The bacterial biota on crustose 120. Nakamura H, Iitaka Y, Kitahara T,
(nonarticulated) coralline algae from Okazaki T, Okami Y. 1977. Structure
Tasmanian waters. Microb. Ecol. of aplasmomycin. J. Antibiot. 30:7 14-
1 1 :221-30 19
1 07 . Lovell FM . 1966. The structure of a 121. Needham J , Andersen R , Kelly MT.
bromine-rich antihiotic. 1. Am. Chern. 1 99 1 . Oncorhyncolide, a novel metabo
Soc. 88:45 1 0-1 1 lite of a bacterium isolated from seawa
108. MacLeod RA. 1965. The question of ter. Tetrahedron Lett. 32 :3 1 5-1 8
the existence of specific marine bacteria. 122. Needham J , Andersen R , Kelly MT.
Bacterial. Rev. 29:9-23 1 992. Biosynthesis of oncorhyncolide,
109. MacLeod RA. 1968. On the role of a metabolite of the seawater bacterial
inorganic ions in the physiology of ma isolate MK157. J. Chem. Soc. Chem.
rine bacteria. In Advances in Microbi Commull. 1367-69
ology of the Sea, ed. MR Droop, EJ 123. Needham J, Kelly MT, Allen TM, Cra
Ferguson Wood, 1 :95-126. London: mer R, Andersen R. 1 994. Cytotoxic
Academic. 239 pp. O-lactone seco hydroxy acids of rhiwxin
1 10. MacLeod RA, Onofrey E. 1956. Nutri analogs isolated from cultures of a ma
tion and metabolism of marine bacteria. rine isolate of Pseudomonas sp.
II. Observations on the relation of (91 V47 ) . J. Antibiot. In press
seawater to the growth of marine bac 124. Needham J, Kelly MT, Ishige M, An
teria. J. Bacterial. 7 1 :661-67 dersen R. 1 994. Andrimid and moi
I ll. Madri PP, Hermel M, Claus G. 1 97 1 . ramides A to C, metabolites produced
The microbial flora o f the sponge in culture by a marine isolate of the
Micraciania pralifera Verril and its bacterium Pseudomonas fluorescens:
ecological implications. Bat. Mar. structure elucidation and biosynthesis.
14 : 1 -5 J. Org. Chem. 59:2058-63
1 12. Makkar NS, Cross T. 1982. Acti 125. Noguchi T, Jeon J-K, Arakawa 0,
noplanetes in soil and on plant litter Sugita H, Deguchi Y, et al. 1986. Oc
from freshwater habitats. 1. Appl. Bac currence of tetrodotoxin and anhydro
terial. 52:209-1 8 tetrodotoxin in Vibrio sp. isolated from
1 13. Marquez MC, Ventosa A , Ruiz-Ber the intestines of a xant hid crab Atergatis
raquero F. 1 990. Marinacaccus hispani floridus. J. Biochem. 99:3 1 1-14
cus, a new species of moderately 1 26. Nolan RD, Cross T. 1988. Isolation and
halophilic Gram-positive cocci. Int. J. screening of actinomycetes. In Acti
Syst. Bacterial. 40: 1 65-69 nomycetes in Biotechnology, ed. M
1 14. Matsui T, Hamada S, Konosu S. 1 98 1 . Goodfellow, ST Cross, M Mordarski,
Difference in accumulation of puffer pp. 1-32. LondonlNew York: Aca
fish toxin and crystalline tetrodotoxin demic. 501 pp.
in the puffer fish, Fugu rubripes ru- 127. Novitsky JA, Morita RY. 1978. Possible
strategy for the survival of marine bac marine bacterium. Tetrahedron Lett. 33:
teria under starvation conditions. Mar. 7663-66
Bioi. 48:289-95 141. Pathirana C, Tapiolas DM, Jensen PR,
128. Novitsky TJ, Kushner J. 1 976. Plano Dwight R, Fenical W. 1 99 1 . Structure
coccus halophilus sp. nov., a faculta determination of maduralide: a new 24-
tively halophilic coccus. Int. l. Syst. membered ring macrolide glycoside
Bacteriol. 26:5 3-57 produced by a marine bacterium (Actin
1 29. Ogata T, Kodama M, Fukuyo Y, Inoue omycetales). Tetrahedron Lett. 32:
T, Karniya H, et al. 1982. The occur 2323-26
rence of Protogonyaulax spp. in Ofunato 142. Paul VJ, ed. 1 992. Ecological Roles of
Bay, in association with the toxification Marine Natural Products. Ithaca: Com
of the scallop Patinop,ecten yessoensis. stock. 245 pp.
Bull. lpn. Soc. Sci. Fish. 48:563-66 143. Pettit GR, Herald CL, Doubek DL, Her
1 30. Oh S, Kogure K, Ohwada K, Simidu ald DL. 1 982. Isolation and structure of
U. 1 99 1 . Correlation between posses bryostatin 1 . l. Am. Chem. Soc.

sion of a respiration-dependent Na' 1 04:6848-49
pump and Na' requirement for growth 144. Pisano MA, Sommer MJ, Brancaccio L.
of marine bacteria. Appl. Environ. Mi 1989. Isolation of bioactive actinomy
crobiol. 57: 1 844-46 cetes from marine sediments using ri
131. Okami Y. 1993. The search for bioactive fampicin. Appl. Microbiol. Biotechnol.
metabolites from marine bacteria. l. 3 1 :609-1 2
Mar. Biotech. 1 :59-65 145. Pisano MA, Sommer MJ, Lopez MM.
1 32. Okami Y, Hotta K, Yoshida M, Ikeda 1 986. Application of pretreatments for
D, Kondo S, Umezawa H. 1 979. New the isolation of bioactive actinomycetes
aminoglycoside antibiotics, istamycins from marine sediments. Appl. Micro
A and B. l. Antibiot. 32:964-66 bioi. Biotechnol. 25:285-88
1 33 . Okami Y, Kurasawa S, Hirose Y. 1980. 146. Porter KG, Feig YS. 1 980. The use of
A new glucanase produced by a marine DAPI for identifying and counting
Bacillus. Agric. Bioi. Chem. 44: 1 19 1-92 aquatic microflora. Limnol. Oceanogr.
1 34. Okami Y, Okazaki T. 1978. Actinomy 25:943-48
cetes in marine environments. In No 147. Rehnstam A-S, Backman S, Smith DC,
cardia and Streptomycetes. ed. M Azam F, Hagstrom A. 1993. Blooms of
Mordarski, W Kurylowicz. W Leljas sequence-specific culturable bacteria in
zewicz, pp. 145-5 1 . Stuttgart: Gustav the sea. FEMS Microbiol. Ecol.
Fisher. 440 pp. 1 02 : 1 61-66
1 35 . Okami Y, Okazaki T, Kitahara T, 148. Reichenbach H, Hofle G. 1993. Biologi
Umezawa H. 1 976. Studies on marine cally active secondary metabolites from
microorganisms. V. A new antibiotic, myxobacteria. Biotech. Adv. 1 1 :219-77
aplasmomycin, produced by a strep 149. Reid RT, Live DH, Faulkner DJ, Butler
tomycete isolated from shallow sea A. 1993. A siderophore from a marine
mud. l. Antibiot. 29: 1 0 1 9-25 bacterium with an exceptional ferric ion
136. Okazaki T, Kitahara T, Okami Y. 1975. affinity constant. Nature 366:455-58
Studies on marine microorganisms. IV. 150. Rinehart KL, Gloer lB, Hughes RG,
A new antibiotic SS-228Y produced by Renis HE, McGovren JP, et aL 1 98 1 .
Chainia isolated from shallow sea mud. Didemnins: antiviral and antitumor dep
l. Antibiot. 28: 176-84 sipeptides from a Caribbean tunicate.
1 37 . Okazaki T. Okami Y. 1975. Actinomy Science 2 12:933-35
cetes tolerant to increased NaCI con 151. Rinehart KL, Shaw PD, Shield LS,
centration and their metabolites. l. Gloer JB, Harbour GC, et al. 1 98 1 . M a
Ferment. Technol. 53:833-40 rine natural products as sources of an
1 38. Pathirana C. Dwight R. Jensen PR, Feni tiviral, antimicrobial, and antineoplastic
cal W, Delgado A, et al. 1 99 1 . Structure agents. Pure Appl. Chem. 53:795-8 1 7
and synthesis of a new butanolide from 1 52. Ritzau M , Keller M, Wessels P, Stetter
a marine actinomycete. Tetrahedron KO, Zeeck A. 1993. New cyclic poly
Lell. 32:7001-4 sulfides from hyperthermophilic Ar
1 39. Pathirana C, Jensen PR, Dwight R, Feni chaea of the Genus Thermococcus.
cal W. 1 992. Rare phenazine L-quino Liebigs Ann. Chem. 8:87 1-76
vose esters from a marine actinomycete. 153. Rosenfeld WD, Zobell C. 1 947. Anti
J. Org. Chem. 57:740-42 biotic production by marine microor
140. Pathirana C. Jensen PR, Fenical W. ganisms. l. Bacteriol. 54:393-98
1 992. Marinone and debromomarinone: 1 54. Roszak DB, Colwell RR. 1987. Survival
antibiotic sesquiterpenoid naphthoqui strategies of bacteria in the natural en
nones of a new structure class from a vironment. Microbiol. Rev. 5 1 :365-79
SECONDARY METABOLITES FROM MARINE B ACTERIA 5 83
1 55 . Rublee RA, Lasker HR, Gottfried M, logical properties. J. Antibiot. 42: 1 556-
Roman MR. 1 980. Production and bac 61
terial colonization of mucus from the 170. Takahashi A, Nakamura H, Kameyama
soft coral Briarium asbestinum. Bull. T, Kurasawa S, Naganawa H, et al.
Mar. Sci. 30:888-93 1987. Bisucaberin, a new siderophore
156. Santavy DL, Willenz P, Colwell RR. sensitizing tumor cells to macrophage
1 990. Phenotypic study of bacteria as mediated cytolysis. II. Physico-chemical
sociated with the Caribbean sclero properties and structure determination.
sponge, Ceratoporella nicholsoni. Appl. J. Alltibiot. 40: 1 67 1 -76
Environ. Microbial. 56: 1750-62 171. Takano R, Imada C, Kamei K, Hara S.
157. Sato K, Okazaki T, Maeda K, Okami 1 9 9 1 . The reactive site of marinostatin,
Y. 1978. New antibiotics, aplasmomy a proteinase inhibitor from marine Al
cins B and C. J. Antibiot. 3 1 :632-35 teronomas sp. B IO-3 1 . J. Biochem.
-
158. Schantz EJ. 1 986. Chemistry and biol

1 10:856-58
ogy of saxitoxin and related toxins. Ann. 172. Takizawa M, Colwell RR, Hill RT.
N. Y. Acad. Sci. 479: 1 5-23 1993. Isolation and diversity of acti
1 58a. Scheuer PJ, ed. 1989. Bioorganic Chem nomycetes in the Chesapeake Bay. Appl.
istry, Vol. 3. LondonlNew York: Sprin Environ. Microbial. 59:997-1002
ger-Verlag. 175 pp. 173. Tapiolas DM, Roman M, Fenical W,
159. Shigemori H, Bae M-A, Yazawa K, Stout TJ, Clardy J. 1991. Octalactins A
Sasaki T, Kobayashi J. 1992. Altermide and B, cytotoxic eight-membered ring
A, a new tetracyclic alkaloid from a lactones from a marine bacterium,
bacterium Altermonas sp. associated Streptomyces sp. J. Am. Chem. Soc. 1 1 3 :
with the marine sponge Halichondria 4682-83
okadai. J. Org. Chem. 57:431 7-20 174. Thompson JE, Walker RP, Faulkner
1 60. Sieburth JM. 1 975. Microbial Sea OJ. 1985. Screening and bioassays for
scapes. Baltimore: Univ. Park Press. 248 biologically-active substances from
pp. forty marine sponge species from San
1 6 1 . Sieburth JM. 1979. Sea Microbes. New Diego, California, USA. Mar. Bioi. 88:
York: Oxford Univ. Press. 491 pp. 1 1-21
1 62. Sieburth JM, Conover JT 1 965. Sar
. 1 75 . Tokuda H, Unemoto T. 1 982. Charac
gassum tannin, an antibiotic which re terization of the respiration-dependent
tards fouling. Nature 208:52-53 Na' pump in the marine bacterium Vi
163. Simidu U, Lee WJ. Kogure K. 1983. brio alginolylicus. J. Bioi. Chern. 257:
Comparison of different techniques for 1 0007-14
determining plate counts of marine bac 176. Toranzo AE, Barja JL, Hetrick FM.
teria. Bull. Jpn. Soc. Sci. Fish. 49: 1 199- 1982. Antiviral activity of antibiotic
1 203 producing marine bacteria. Can. J. Mi
164. Simidu U, Noguchi T, Hwang 0, Shida crobial. 28:231-38
Y, Hashimoto K. 1987. Marine bacteria 177. Tresner HD, Hayes JA, Backus EJ.
which produce tetrodotoxin. Appl. En 1968. Differential tolerance of strep
viron. Microbiol. 53: 1 7 1 4- 1 5 tomycetes to sodium chloride as a taxo
1 65. Stierle AC, Cardellina J H II, Singleton nomic aid. Appl. Microbial. 1 6 : 1 1 34-36
FL. 1988, A marine Micrococcus pro 178. Trischman J, Jensen PR, Fenical W.
duces metabolites ascribed to the sponge 1 994. Halobacilin: a cytotoxic cyclic
Tedania ignis. Experientia 44: 1021 acylpeptide of the iturin class produced
166. Stierle DB, Stierle AA. 1992. Pseudo by a marine Bacillus sp. Tetrahedron
monic acid derivatives from a marine Lett. In press
bacterium. Experientia 48: 1 1 65-69 179. Trischman J, Tapiolas OM, Fenical W.
1 67 . Suffness M, Newmann OJ, Snader K. Dwight R, Jensen PR, et al. 1 994. Sali
1989 . Discovery and development of namide A and B anti-inflammatory dep
antineoplastic agents from natural sipeptides from a marine streptomycete.
sources. See Ref. 158a, 3 : 1 3 1-68 J. Am. Chern. Soc. 1 16:757-58
168. Takahashi A, Ikeda D, Nakamura H, 180. Umezawa H , Okami Y, Kurasawa S,
Naganawa H, Kurasawa S, et al. 1989. Ohnuki T, Ishizuka M, et al. 1983.
Altemicidin, a new acaricidal and anti Marinactan, antitumor polysaccharide
tumor substance. II. Structure determi produced by marine bacteria. J. Anribiot.
nation. J. Antibiol. 42: 1 562-66 36:47 1 -77
1 69. Takahashi A, Kurosawa S, Ikeda D, 181. Unson MD. Faulkner OJ. 1993. Cyano
Okami Y, Takeuchi T. 1989. Altemi bacterial symbiont biosynthesis of chlo
cidin, a new acaricidal and antitumor rinated metabolites from Dysidea
substance. I. Taxonomy, fermentation, herbacea (Porifera). Experienria 49:
isolation and physico-chemical and bio- 349-53
1 82. Uriz MJ, Martin D, Turon X, Ballesteros 192. Weyland H, Helmke E. 1988. Acti
E, Hughes R, Acebal C. 199 1 . An ap nomycetes in the marine environment.
proach to the ecological significance of In The Biology of Actinomycetes '88.
chemically mediated bioactivity in Proceedings of the 7'" International
Mediterranean benthic communities. Symposium on the Biology of Actinomy
Mar. Ecoi. Prog. Ser. 70:175-88 cetes, ed.Y Okami, T Beppu, H
1 83. Vrolijk NH, Targett NM, Baier RE, Ogawara, pp. 294-99. Tokyo: Jpn. Sci.
Meyer AE. 1990. Surface characteristics Soc. 508 pp.
of two gorgonian soft coral species: 193. Wilkinson CR. 1978. Microbial associa
implications for a natural antifouling tions in sponges. II. Numerical analysis
defense. Biofouling 2:39-54 of sponge and water bacterial popula
184. Wahl M, Jensen PR, Fenical W. 1 994. tions. Mar. Bioi. 49:169-76
Chemical control of bacterial epibiosis 194. Wilkinson CR. 1987. Significance of
on ascidians. Mar. Eco/. Prog. Ser. In
microbial symbionts in sponge evolution

press and ecology. Symbiosis 4: 135-46
185. Walker JP, Colwell RR. 1975. Factors 195. Wilkinson CR, Nowak M, Austi n B,
affecting enumeration and isolation of Colwell RR. 198 1 . Specificity of bac

actinomycetes from Chesapeake Bay terial symbionts in Mediterranean and
and southeastern Atlantic Ocean sedi Great Barrier Reef sponges. Microb.
ments. Mar. Bioi. 30:193-201 Ecol. 7 : 1 3-21
1 86. Walls JT, Ritz DA, Blackman MJ. 1993. 1 96. Wisse GA, MacLeod RA. 1989. Role
Fouling, surface bacteria and antibacte of Na+ in growth, respiration and mem
rial agents of four bryozoan species brane transport in the marine bacterium
found in Tasmania, Australia. Exp. Mar. Pseudomonas doudoroffii 70. Arch. Mi
Bio/. Ecol. 169:1-13 crobiol. 153:64-7 1
1 87. Wardell IN, Brown CM, Flannagan B . 197. Wood EJF. 1952. The micrococci in a
1 983. Microbes and surfaces. I n Mi marine environment. J. Gen. Microbiol.
crobes in Their Natural Environment, 6:205-10
ed. CJH Slater, R Whittenbury, JWT 198. Wratten SJ, Wolfe MS, Andersen RJ,
Wimpenny, pp. 350--7 8. London: Cam Faulkner DJ. 1 977. Antibiotic meta
bridge Univ. Press. 497 pp. bolites from a marine pseudomonad.
188. Waterbury JB, Calloway CB, Turner A ntimicrob. Agents Chemother. 1 1 :
RD. 1983. A cellulolytic nitrogen-fixing 4 1 1-14
bacterium cultured from the gland of 199. Yasumoto T, Yasumura D, Yotsu M,
Deshayes in shipworms (Bivalvia: Michishita T, Endo A, Kotaki Y. 1 986.
Teredinidae). Science 221: 1401-3 Bacterial production of tetrodotoxin and
1 89. Waterbury JB, Watson SW, Guillard anhydrotetrodotoxin. Agric. BioI. Chem.
RLL, Brand LE. 1979. Widcsprcad oc 50:793-95
currence of a unicellular, marine, plank 200. Yotsu M, Yamazaki T, Meguro Y, Endo
tonic cyanobacterium. Nature 227: A, Murata M, et a!. 1987. Production
293-94 of tetrodotoxin and its derivatives by
190. Wei SL, Young RE. 1989. Development Pseudomonas sp. isolated from the skin
of symbiotic bacterial bioluminescence of a pufferfish. Toxicon 25:225-28
i n a nearshore ceph alopod, Euprymna 201 . ZoBell CEo 1946. Marine Microbiology.
scolopes. Mar. Bioi. 103:541-46 Waltham, MA: Chronica Botanica. 240
191. Weyland H. 1 98 1 . Characteristics of pp.
actinomycetes isolated from marine 202. ZoBell CE, Upham HC. 1944. A list of
sediments. Zentralbl. Bakteriol. Para marine bacteria including descriptions
sitenkd. Infektionskr. 1 l : 309-14 of sixty new species. Bull. Scripps Inst.
(Supp!.) Oceanogr. 5:239-92

Jensen. 1994, Strategies For The Discovery of Secondary Metabolites PDF

Caricato da

Informazioni sul documento

Titolo originale

Copyright

Formati disponibili

Condividi questo documento

Condividi o incorpora il documento

Opzioni di condivisione

Hai trovato utile questo documento?

Questo contenuto è inappropriato?

Copyright:

Formati disponibili

Jensen. 1994, Strategies For The Discovery of Secondary Metabolites PDF

Caricato da

Copyright:

Formati disponibili

ANNUAL

STRATEGIES FOR THE

Paul R. Jensen and William Fenical

ECOLOGICAL CONSIDERATIONS ...... .... . .. . . ... . . .... . .. . . . .. . . .. . . . . 561

BIOLOGICAL ACTIVITIES AND NOVEL ORGANIC SUBSTANCES............ 574

CONCLUDING REMARKS. . . . . . . . .. . ..... . . .. . . . .. . . . .... . . . . . . . . . . . . .. . . 576

Marine microorganisms have become an important point of study in the search

biosynthetic pathways of terrestrial plants and microorganisms. In the late

of the biomedical potential and pharmacological properties of marine metabo­

study of marine bacteria, a logical and potentially productive avenue of re­

trial bacteria and the renewable nature of culturable resources.

Table 1 Novel organic molecules reported from marine bacteria

Producing strain Source Biomedical activity References

Chaina purpurogena Sediment Antibiotic, anticancer 86, 136

Actinomycete Sediment No activity reported 138

Actinomycete Gorgonian soft coral Anticancer, antibiotic 173

Pseudomonas bromoutilis (Alteromo- Seagrass Antibiotic 18. 107

undescribed s econdary metabolites, and if so, are environmental factors spe­

by a nonmarine strain. However, if this substance is produced only when

research, a better understanding of marine bacterial ecology is essential. This

Bacterial taxonomic and metabolic diversity are fundamentally interrelated

a correlation between taxonomic and biosynthetic diversity has yet to be

natural products research still represents a rational approach to novel metabo­

The Distribution of Bacteria in the Sea

In the course of isolating marine bacteria for chemical evaluation, we have

The Associations of Bacteria with Plants and Invertebrates

symbionts from transient associates, and the role of symbiotic bacteria in

that the majority of marine organisms have specific bacterial populations

reported in greater abundance in sediments (118), the relative abundance of

The Role of Symbiotic Bacteria in Secondary Metabolite

tant data in support of bacterial b iosynthesis.

cytometry to separate cyanobacterial symbionts (Oscillatoria spongeliae) from

Effects of Marine MetaboLites upon the Distribution of Bacteria

separate reports, investigators proposed that antibiotic algal phenols were

A lthough the in situ microbiological effects of marine natural products

distribution of bacteria on living marine surfaces. One example comes from

The Adaptations of Bacteria to Diverse Marine Habitats

to diverse marine habitats provide seemingly limitless evolutionary opportu­

cludes the recognition that a gene (ompH) from a gram-negative bacterium

it is not surprising that large numbers of bacteria appear to be unculturable.

The Distinction Between Marine and Terrestrial Bacteria

generation of ATP. Oh et al (130) have shown that the sodium requirement in

positive coccus Gaffkya homari (now Aerococcus viridans), the pathogenic

BIOLOGICAL ACTIVITIES AND NOVEL ORGANIC

The ability of marine bacteria to produce antibacterial substances was docu­

discovery (because of low hit rates) and to effective bioassay-guided chemical

We mentioned that novel secondary metabolites have been isolated from a

genera, including Moraxella (52) and Alcaligenes ( 1 6) produce biologically

Today, marine bacteria are being recognized as an important resource for

ation of this manuscript. In addition, we thank Carole Bewley for helpful

1. AI-Ogily SM, Knight Jones EW. 1977.

croorganisms: the antibacteri al spec­ tions and potential for biotechnology .

1966. Production of a pyrrole antibiotic Identification of deep-sea-sediment bac­

tive Metabolites from Microorganisms, monas agent isolated from a marine

39 VV. Mal'tsev II, Voinov VG, Fedoreyev

Goericke R, Waterbury ]B, Welsch­ from a marine bacterium associated with

W Harder, K Schleifer, 2: 1 769-84. New 39. Fenical W. 1993. Chemical studies of

by manne actinomycete. l. Antibiot. tion of paralytic shellfish toxins by a

U. 1 99 1 . Correlation between posses­ bryostatin 1 . l. Am. Chem. Soc.

158. Schantz EJ. 1 986. Chemistry and biol­

microbial symbionts in sponge evolution

affecting enumeration and isolation of Colwell RR. 198 1 . Specificity of bac­

Potrebbero piacerti anche

of the biomedical potential and pharmacological properties of marine metabo

study of marine bacteria, a logical and potentially productive avenue of re

undescribed s econdary metabolites, and if so, are environmental factors spe

natural products research still represents a rational approach to novel metabo

to diverse marine habitats provide seemingly limitless evolutionary opportu

The ability of marine bacteria to produce antibacterial substances was docu

croorganisms: the antibacteri al spec tions and potential for biotechnology .

1966. Production of a pyrrole antibiotic Identification of deep-sea-sediment bac

Goericke R, Waterbury ]B, Welsch from a marine bacterium associated with

U. 1 99 1 . Correlation between posses bryostatin 1 . l. Am. Chem. Soc.

158. Schantz EJ. 1 986. Chemistry and biol

affecting enumeration and isolation of Colwell RR. 198 1 . Specificity of bac