Hybrid isoprenoid secondary metabolite production in terrestrial and marine actinomycetes

Terpenoids are among the most ubiquitous and diverse secondary metabolites observed in nature. Although actinomycete bacteria are one of the primary sources of microbially derived secondary metabolites, they rarely produce compounds in this biosynthetic class. The terpenoid secondary metabolites that have been discovered from actinomycetes are often in the form of biosynthetic hybrids called hybrid isoprenoids (HIs). HIs include significant structural diversity and biological activity and thus are important targets for natural product discovery. Recent screening of marine actinomycetes has led to the discovery of a new lineage that is enriched in the production of biologically active HI secondary metabolites. These strains represent a promising resource for natural product discovery and provide unique opportunities to study the evolutionary history and ecological functions of an unusual group of secondary metabolites.     

Biologically active oxylipins from seaweeds

Our previous research has shown that many red algae metabolize polyunsaturated fatty acids to oxidized products resembling the eicosanoid hormones from mammals. We have extended these studies to members of the Phaeophyceae and Chlorophyta and find they also possess similar biosynthetic pathways. From several we have identified novel prostaglandin-like substances. Studies of the molecular mechanisms by which some of these marine oxylipins are formed have revealed that novel oxidative reactions are utilized. Understanding of these biosynthetic pathways in detail has allowed their utilization to produce research biochemicals of high value, such as 12S-hydroperoxyeicosatetraenoic acid (12S-HPETE). Because of their biological properties, seaweed-derived oxylipins have potential utility as pharmaceuticals and research biochemicals.     

Symbiotic and dietary marine microalgae as a source of bioactive molecules–experience from natural products research

Cyanobacterial metabolites have proven to be invaluable as tools in thedissection of signal transduction pathways in mammalian cells and some arecurrently under clinical evaluation as drug candidates. It is now also realizedthat cyanobacteria are the true biosynthetic origin of many bioactive moleculesisolated from marine invertebrates; marine invertebrates may sequestercyanobacteria through diet or by symbiosis. This review discusses thedietary-derived cyanobacterial origin of the dolastatins, potent cytotoxiccompounds, originally isolated from the Indian Ocean sea hare,Dolabella auricularia. A discussion on the dietarydissemination of cyanobacterial metabolites through the marine food chain isalso presented. Reference to the metabolites isolated fromDysidea sponges is given to illustrate their origin fromsymbiotic cyanobacteria associated with this organism.     

Photoactivated perylenequinone toxins in fungal pathogenesis of plants

Several genera of plant pathogenic fungi produce photoactivated perylenequinone toxins involved in pathogenesis of their hosts. These toxins are photosensitizers, absorbing light energy and generating reactive oxygen species that damage the membranes of the host cells. Studies with toxin-deficient mutants and on the involvement of light in symptom development have documented the importance of these toxins in successful pathogenesis of plants. This review focuses on the well studied perylenequinone toxin, cercosporin, produced by species in the genus Cercospora. Significant progress has been made recently on the biosynthetic pathway of cercosporin, with the characterization of genes encoding a polyketide synthase and a major facilitator superfamily transporter, representing the first and last steps of the biosynthetic pathway, as well as important regulatory genes. In addition, the resistance of Cercospora fungi to cercosporin and to the singlet oxygen that it generates has led to the use of these fungi as models for understanding cellular resistance to photosensitizers and singlet oxygen. These studies have shown that resistance is complex, and have documented a role for transporters, transient reductive detoxification, and quenchers in cercosporin resistance.     

Biotechnological production of eicosapentaenoic acid: From a metabolic engineering point of view

Eicosapentaenoic acid (EPA) is an ω3 polyunsaturated fatty acid which has been demonstrated to play important roles in a number of aspects of human health. EPA is traditionally obtained from marine fish oils. However, the shrinking fish populations are making the sustainability of these sources questionable. Consequently, alternative sources of EPA are being sought, especially from marine microalgae, bacteria, and fungi. These microorganisms contain relatively large amounts of high-quality EPA and they are the primary producers of this important fatty acid. There are two distinct pathways for EPA de novo biosynthesis in microbial systems: the desaturation and elongation pathway and the polyketide pathway. Genes involved in the biosynthetic pathways have been identified from different microorganisms and characterized in depth. In addition, numerous strategies have been developed for commercial production of EPA by microbial fermentation, among which strain improvements by genetic engineering could provide high-yield producers of EPA. In this review, we summarize recent efforts and experiences devoted to metabolic engineering of various microorganisms that lead to efficient biocatalysts for the production of EPA, as well as the key limitations and challenges. The combination of traditional biochemistry and molecular biology with new systems biology and synthetic biology tools will provide a better view of EPA biosynthesis and a greater potential of microbial production. Continued advances in metabolic engineering will help to improve the final titer, productivity, and yield of EPA.     

源于微生物的海鞘天然产物

海洋动物是具有生物活性海洋天然产物的重要来源。海鞘中含有丰富的微生物类群,如细菌、放线菌、真菌和蓝细菌。越来越多的直接或间接证据表明,一些从海鞘中分离的天然产物并不是海鞘本身产生的,而是由其共生微生物产生的。本文对近些年来的海鞘天然产物的微生物来源的研究方法进行综述,包括可培养细菌的分离、不可培养细菌的粗提物检测、宏基因组学、全基因组测序等直接方法,以及化合物结构比对的间接方法。通过对海鞘-微生物共生体中天然产物生物合成来源的研究,不仅可以从根本上解决动物药源的问题,而且可为研究海鞘与微生物共生关系提供有力证据。     

海洋共附生微生物天然产物生物合成基因研究进展

对海洋无脊椎动物天然产物的研究表明,很多种活性物质的真正生产者是与其共生或附生的未培养微生物.克隆这些未培养微生物中特定活性物质的生物合成基因,不仅为活性物质的来源提供遗传学证据,也使通过异源表达相关生物合成基因来大量获取目标化合物成为可能.本文综述了来源于海绵、海鞘、苔藓虫、深海管状蠕虫和深海沉积物中共附生微生物天然产物生物合成基因簇的研究进展.     

Manzamines

Manzamines are a unique class of β-carboline marine alkaloids with an unusual tetra- or pentacyclic system. These alkaloids have shown a variety of bioactivities against infectious diseases, cancer and inflammatory diseases. The greatest potential for the manzamine alkaloids appears to be against malaria, with improved potency relative to chloroquine and artemisinin. Over 80 manzamine-related alkaloids have been isolated from more than 16 species of marine sponges belonging to five families distributed from the Red Sea to Indonesia, which suggests a possible microbial origin for manzamine alkaloids. The current review summarizes marine literature, focusing on the biological activities of manzamines, the possible microbial origin of this class of compounds and the Red Sea as a possible source of manzamines from biosynthetic gene clusters of Red Sea microbes.     

The Historical Factor: The Biosynthetic Relationships Between Amino Acids and Their Physicochemical Properties in the Origin of the Genetic Code

Two forces are in general, hypothesized to have influenced the origin of the organization of the genetic code: the physicochemical properties of amino acids and their biosynthetic relationships. In view of this, we have considered a model incorporating these two forces. In particular, we have studied the optimization level of the physicochemical properties of amino acids in the set of amino acid permutation codes that respects the biosynthetic relationships between amino acids. Where the properties of amino acids are represented by polarity and molecular volume we obtain indetermination percentages in the organization of the genetic code of approximately 40%. This indicates that the contingent factor played a significant role in structuring the genetic code. Furthermore, this result is in agreement with the genetic code coevolution hypothesis, which attributes a merely ancillary role to the properties of amino acids while it suggests that it was their biosynthetic relationships that organized the code. Furthermore, this result does not favor the stereochemical models proposed to explain the origin of the genetic code. On the other hand, where the properties of amino acids are represented by polarity alone, we obtain an indetermination percentage of at least 21.5%. This might suggest that the polarity distances played an important role and would therefore provide evidence in favor of the physicochemical hypothesis of genetic code origin. Although, overall, the analysis might have given stronger support to the latter hypothesis, this did not actually occur. The results are therefore discussed in the context of the different theories proposed to explain the origin of the genetic code. Received: 10 September 1996 / Accepted: 3 March 1997     

赤潮藻毒素危害及其检测方法研究进展

近年来,世界各地包括我国部分沿海地区赤潮发生过度频繁,许多赤潮藻毒素不但使其他海洋生物中毒,对人类健康也有潜在的危害,因此对赤潮藻毒素的研究已经成为海洋环境科学研究的重要内容。我们介绍了赤潮藻毒素的危害,并详细综述了赤潮藻毒素检测技术研究进展。     

Human insulin

The two human insulins of clinical importance are (a) semisynthetic human insulin prepared from pork pancreas by enzymatically substituting threonine for alanine-the last amino acid in the beta chain-thereby transforming pork insulin in vitro to human insulin; and (b) biosynthetic human insulin synthesized biotechnologically in Escherichia coli-K12. Using this latter technique, it is possible to produce mass quantities of highly purified insulin for the treatment of insulin-dependent diabetics, avoiding the problems inherent in supplies of insulin produced from animal pancreas. It has been suggested that to avoid confusion the two human insulins should be called semisynthetic human insulin of pork origin and biosynthetic human insulin of E. coli origin, respectively. These insulins have four advantages over highly purified animal insulins: (a) they induce lower titers of circulating insulin antibodies; (b) their subcutaneous injection is associated with fewer skin reactions; (c) they are absorbed more rapidly from the injection site; and (d) less degradation occurs at the site of injection. These data indicate that newly diagnosed insulin-dependent diabetes, particularly in children, should be treated with either of the two human insulins. The warranty against inadequate supplies of insulin offered by biosynthetic human insulin makes the use of pork insulins unnecessary and beef insulins totally useless.     

Cloning of Complementary and Genomic DNAs Encoding Echotoxins,Proteinaceous Toxins from the Salivary Gland of Marine Gastropod <Emphasis Type="Italic">Monoplex echo</Emphasis>

Echotoxins are hemolytic and lethal proteinaceous toxins of about 25 kDa that are contained in the salivary gland of marine gastropod Monoplex echo. In this study, complementary DNAs (cDNAs) encoding four echotoxins (2, A, B1 and B2) were isolated from the cDNA library constructed from the M. echo salivary gland and completely sequenced. Although the amino acid sequence identity between the four echotoxins and actinoporins (20 kDa hemolysins from sea anemones) was very low (12–16%), some amino acid residues important for the biological activity of actinoporins were well conserved in the echotoxins. In the case of echotoxin 2, the genomic DNA corresponding to the coding region was amplified. Sequencing data revealed that the echotoxin 2 gene is devoid of introns within the coding region as reported for the actinoporin genes. These results suggest that echotoxins have evolved from actinoporins or both toxins have evolved from the same ancestor.     

Photosynthetic marine organisms as a source of anticancer compounds

Since early human history, plants have served as the most important source of medicinal natural products, and even in the “synthetic age” the majority of lead compounds for pharmaceutical development remain of plant origin. In the marine realm, algae and seagrasses were amongst the first organisms investigated by marine natural products scientists on their quest for novel pharmaceutical compounds. Forty years after the pioneering work in the field of marine drug discovery began, the biodiversity of marine organisms investigated as potential sources of anticancer, anti-inflammatory, and antibiotic compounds has increased tremendously. Nonetheless, marine plants are still an important source of novel secondary metabolites with interesting biomedical properties. The present review focuses on the antitumour properties of compounds isolated from marine algae, phytoplankton, mangroves, seagrasses, or cordgrasses. Compounds produced by marine epi- or endophytic fungi are also discussed.     

The Level and Landscape of Optimization in the Origin of the Genetic Code

We consider a model of the origin of genetic code organization incorporating the biosynthetic relationships between amino acids and their physicochemical properties. We study the behavior of the genetic code in the set of codes subject both to biosynthetic constraints and to the constraint that the biosynthetic classes of amino acids must occupy only their own codon domain, as observed in the genetic code. Therefore, this set contains the smallest number of elements ever analyzed in similar studies. Under these conditions and if, as predicted by physicochemical postulates, the amino acid properties played a fundamental role in genetic code organization, it can be expected that the code must display an extremely high level of optimization. This prediction is not supported by our analysis, which indicates, for instance, a minimization percentage of only 80%. These observations can therefore be more easily explained by the coevolution theory of genetic code origin, which postulates a role that is important but not fundamental for the amino acid properties in the structuring of the code. We have also investigated the shape of the optimization landscape that might have arisen during genetic code origin. Here, too, the results seem to favor the coevolution theory because, for instance, the fact that only a few amino acid exchanges would have been sufficient to transform the genetic code (which is not a local minimum) into a much better optimized code, and that such exchanges did not actually take place, seems to suggest that, for instance, the reduction of translation errors was not the main adaptive theme structuring the genetic code.     

Marine yeasts as biocontrol agents and producers of bio-products

As some species of marine yeasts can colonize intestine of marine animals, they can be used as probiotics. It has been reported that β-glucans from marine yeast cells can be utilized as immuno-stimulants in marine animals. Some siderophores or killer toxins produced by marine yeasts have ability to inhibit growth of pathogenic bacteria or kill pathogenic yeasts in marine animals. The virulent factors from marine pathogens can be genetically displayed on marine yeast cells, and the yeast cells displaying the virulent factors can stimulate marine animals to produce specific antibody against the pathogens. Some marine yeast cells are rich in proteins and essential amino acids and can be used in nutrition for marine animals. The marine yeast cells rich in lipid can be used for biodiesel production. Recently, it has been reported that some strains of Yarrowia lipolytica isolated from marine environments can produce nanoparticles. Because many marine yeasts can remove organic pollutants and heavy metals, they can be applied to remediation of marine environments. It has been shown that the enzymes produced by some marine yeasts have many unique properties and many potential applications.     

Microalgal metabolites

The extraordinary chemical diversity seen in the cyanobacteria (blue-green algae) is especially pronounced in the ubiquitous tropical marine species, Lyngbya majuscula. The gene clusters responsible for the production of some of the secondary metabolites have recently been elucidated. The dinoflagellates, which are lower eukaryotic algae, also demonstrate chemical diversity and produce unique polycyclic ethers of polyketide origin. A new mechanism for the formation of the truncated polyketide backbones has recently been proposed. The toxicogenicity of dinoflagellates of the genus Pfiesteria has been the focus of controversy--are they 'killer organisms', as alleged? A recent investigation of Pfiesteria genes seems to rule out the presence of polyketide synthase, which is the gene responsible for the production of most dinoflagellate toxins.     

Was macht Molche giftig?

What makes newts toxic? Tetrodotoxin is one of the strongest toxins of natural origin, which specifically blocks voltage-gated sodium channels of nerve and muscle. It was first detected in marine puffer fish (Tetraodontidae), but later also in other marine and terrestrial animals such as in amphibians. Particularly, some newt species (Urodela) of the New and Old World exhibit extremely high concentrations of the toxin which, to some extent, provide protection against predators. However, North American garter snakes of the genus Thamnophis proved to be toxin-resistant which is based on amino acid substitutions at the toxin-binding site of the sodium channel. The origin of the toxin is under discussion. Apparently, it is not synthesized by the newts but is most likely acquired via the food-chain and stored. Moreover, other amphibians such as some toads and frogs have been found to contain tetrodotoxin. The significance of the toxin for survival of species and its interweaving with the challenges of the ecosystems is still unclear.     

Microalgae metabolites: A rich source for food and medicine

Microalgae are one of the important components in food chains of aquatic ecosystems and have been used for human consumption as food and as medicines. The wide diversity of compounds synthesized from different metabolic pathways of fresh and marine water algae provide promising sources of fatty acids, steroids, carotenoids, polysaccharides, lectins, mycosporine-like amino acids, halogenated compounds, polyketides, toxins, agar agar, alginic acid and carrageenan. This review discusses microalgae used to produce biological substances and its economic importance in food science, the pharmaceutical industry and public health.     

The zebrafish (Danio rerio) embryo as a model system for identification and characterization of developmental toxins from marine and freshwater microalgae

The zebrafish (Danio rerio) embryo has emerged as an important model of vertebrate development. As such, this model system is finding utility in the investigation of toxic agents that inhibit, or otherwise interfere with, developmental processes (i.e. developmental toxins), including compounds that have potential relevance to both human and environmental health, as well as biomedicine. Recently, this system has been applied increasingly to the study of microbial toxins, and more specifically, as an aquatic animal model, has been employed to investigate toxins from marine and freshwater microalgae, including those classified among the so-called "harmful algal blooms" (HABs). We have developed this system for identification and characterization of toxins from cyanobacteria (i.e. "blue-green algae") isolated from the Florida Everglades and other freshwater sources in South and Central Florida. Here we review the use of this system as it has been applied generally to the investigation of toxins from marine and freshwater microalgae, and illustrate this utility as we have applied it to the detection, bioassay-guided fractionation and subsequent characterization of developmental toxins from freshwater cyanobacteria.     

Bryostatins: biological context and biotechnological prospects

Bryostatins are a family of protein kinase C modulators that have potential applications in biomedicine. Found in miniscule quantities in a small marine invertebrate, lack of supply has hampered their development. In recent years, bryostatins have been shown to have potent bioactivity in the central nervous system, an uncultivated marine bacterial symbiont has been shown to be the likely natural source of the bryostatins, the bryostatin biosynthetic genes have been identified and characterized, and bryostatin analogues with promising biological activity have been developed and tested. Challenges in the development of bryostatins for biomedical and biotechnological application include the cultivation of the bacterial symbiont and heterologous expression of bryostatin biosynthesis genes. Continued exploration of the biology as well as the symbiotic origin of the bryostatins presents promising opportunities for discovery of additional bryostatins, and new functions for bryostatins.