Seriniquinone is a natural quinone isolated from a rare marine bacterium of the genus Serinicoccus. This secondary metabolite has been shown to have anticancer properties, which has raised attention of the scientific community. In this short report, we present the first investigation of the gas-phase chemistry fragmentation reactions of seriniquinone in electrospray ionization tandem mass spectrometry (ESI-MS/MS), to be further applied in pharmacokinetics and metabolism studies. All the proposals herein were supported by computational chemistry.

The actinomycete strain BRA 177 was recovered from sediment samples collected at the St. Peter and St. Paul Archipelago, Brazil. This work accessed the ability of this strain, identified as Actinomadura sp., to produce bioactive metabolites by exploring the genome and characterizing chemistry and cytotoxicity of isolated compounds. From the crude ethyl acetate extract, the pigments nonylprodigiosin, cyclononylprodigiosin and methylcyclooctilprodigiosin were isolated and displayed cytotoxicity against human tumor and non-tumor cell lines. Sequencing, assembling and prospection of BRA 177 draft genome led to identification of two contigs encoding enzymes with high homology to those from prodiginine biosynthetic gene clusters (BGC) in actinomycetes. Further, Actinomadura sp. BGC presented unique putatives RedJ thioesterase and RedL-like type I PKS, involved on selection of prodiginine biosynthetic fatty acyl precursor, and RedG-like Rieske oxygenase, key for cyclization of the prodiginines, suggesting that cyclononilprodigiosin and methylcyclooctylprodigiosin could actually be considered chemical signatures of Actinomadura spp.

Abstract Dinoflagellates are an important source of unique bioactive secondary metabolites. Symbiotic species, commonly named zooxanthellae, transfer most of their photosynthetically fixed carbon to their host. The mutualistic relationship provides the organic metabolites used for energy production but there are very few reports of the role of the dinoflagellates in the production of secondary metabolites in the symbiotic association. Corals and other related cnidarians are the most well-known animals containing symbiotic dinoflagellates. In the present paper we describe the isolation of amphidinolide P (1) from the octocoral Stragulum bicolor and its prey, the nudibranch Marionia limceana, collected off the coasts of Fortaleza (Ceará, Brazil). The coral extracts also contained 3-O-methyl derivative (2) of amphidinolide P, together with minor compounds still under investigation. Amphidinolides have been so far reported only in laboratory cultures of Amphidinium sp., thus compounds 1 and 2 represents the first identification of these polyketides in invertebrates. The finding proves the possibility to isolate amphidinolides from a natural symbiosis, enabling further biological and biotechnological studies.

Abstract Four bisabolanes 1–4, including perezone (1) and triacetyl perezone (2), were isolated through a bioassay-guided fractionation of the extract obtained from the Caribbean gorgonian coral Pseudopterogorgia rigida collected during an expedition cruise to the Bahamas. All isolated compounds showed to be cytotoxic toward panel of four human tumor cell lines, as quantified by the MTT assay after 72 h incubation. Perezone (1), the most active one, was further analyzed, showing to be cytotoxic, but not selective, in a 12-cell line panel comprising tumor and non-tumor, as well as human and murine cells. Additionally, 1 was assayed for cytotoxicity against HL-60 leukemic cells. Pre-treatment with an acute free radical scavenger (L-NAC) before exposure of cells to perezone virtually eliminated the generation of intracellular ROS and lessened its severe cytotoxicity. The protective effect delivered by L-NAC evidences that the mechanism of perezone-induced cytotoxicity is partially associated to production of ROS and a consequent induction of oxidative stress.

Previous studies demonstrated that the crude extract of the ascidian Eudistoma vannamei, endemic from northeasttern Brazil, strongly hinders growth of tumor cells in vitro by inducing apoptosis due to tryptophan derivatives, which are commonly found in bacteria. This study presents a bioactivity-guided screening among actinomycetes, associated with E. vannamei, aiming at recognizing active principles with biological relevance. Twenty strains of actinomycetes, designated as EVA 0101 through 0120, were isolated from colonies of E. vannamei among which 11 were selected for cytotoxicity evaluation. The extracts from EVA 0102, 0103, 0106, 0109 and 0113 were the most active, and were further studied for IC50 determination and chemical analysis by ¹H NMR. IC50 values obtained ranged from 3.62 µg mL-1 (for EVA 0109 in leukemia cells) to 84.65 µg/mL (for EVA 0106 in melanoma cells). All active extracts exhibited the same TLC and spectroscopic profiles, suggesting the presence of quinones and other related secondary metabolites. Furthermore, these strains were identified and compared based on their respective 16S rRNA sequences. The results herein identified the five strains as Micromonospora spp. while phylogenetic analysis suggests that they are possibly two different Micromonospora species producing the cytotoxic compounds.

Estudios previos demostraron que el extracto crudo de la ascidia Eudistoma vannamei, endémica de la costa noreste de Brasil, obstaculiza fuertemente el crecimiento de células tumorales in vitro por inducir apoptosis. El análisis químico del extracto sugirió la presencia de derivados de triptófano, comúnmente encontrados en bacterias. El estudio presenta un screening de citotoxicidad en actinomicetos asociados con E. vannamei, para reconocer principios activos biológicamente relevantes. Veinte cepas de actinomicetos, designados como EVA 0101 hasta 0120, fueron aisladas de colonias de E. vannamei y 11 fueron seleccionadas para la evaluación de citotoxicidad. Los extractos de EVA 0102, 0103, 0106, 0109 y 0113 resultaron las más activas y fueron estudiadas para determinación de la CL50 y perfiles de CCF y ¹H RMN. Las CL50 obtenidas oscilaron entre 3,62 (EVA 0109 en las células de leucemia) y 84,65 µg mL-1 (EVA 0106 en las células de melanoma). Los extractos activos presentan el mismo perfil de CCF y espectroscópico, lo que sugiere la presencia de quinonas y metabolitos secundarios relacionados. Además, las cinco cepas fueron identificadas y comparadas sobre la base de sus secuencias de 16S ARNr. Las que se identificaron como Micromonospora spp. y el análisis filogenético sugiere que sean por lo menos dos especies de Micromonospora las que producen los compuestos citotóxicos.

O estudo fitoquímico do invertebrado marinho Eudistoma vannamei levou ao isolamento de derivados do alcalóide estaurosporina, que apresentaram uma potente atividade citotóxica contra linhagens de células humanas de câncer. A ocorrência destes alcalóides pode ser correlacionada a presença de microorganismos associados ao animal, como por exemplo bactérias do gênero Streptomyces. Visando confirmar essa hipótese, precedeu-se o isolamento e cultivo dos microrganismos associados com E. vannamei (84 cepas isoladas). Estudos sistemáticos das reações de fragmentação da estaurosporina e derivados, apoiados pela química computacional, permitiram definir os principais padrões de fragmentação. Esses padrões foram utilizados em análises exploratórias por LC-MS/MS visando pesquisar a presença de derivados de estaurosporinas nos microrganismos isolados. Esse procedimento permitiu identificar estaurosporina em uma espécie de Streptomyces, contudo não com o mesmo padrão de oxidação da molécula ativa isolada no extrato da Ascídia. Finalmente, o extrato dessa cepa mostrou também significativa atividade citotóxica contra linhagens de câncer.

Former bioactivity-guided analysis of the marine invertebrate Eudistoma vannamei led to the isolation of staurosporine derivatives, which revealed strong cytotoxic activity against several human cancer cell lines. The occurrence of such alkaloids in E. vannamei may be correlated to the presence of associated biota, such as Streptomyces bacteria. In agreement to this hypothesis, marine microorganisms associated with E. vannamei were recovered and cultured, leading to a total of 84 isolated bacterial strains. Gas phase fragmentation reactions of staurosporine and derivatives were systematically studied and the analyzed results further supported by computational chemistry studies. The resulting fragment patterns were used to search for the presence of different derivatives in extracts of isolated microorganisms, thereby using LC-MS/MS analysis in MRM mode. These results evidenced that one isolated Streptomyces sp. was able to generate staurosporine, while none of the hydroxy-7-oxo derivatives were detected. Finally, significant cytotoxic activity against human cancer lines was observed for one of the extracts.

Um método de espectrometria de massas sequencial com ionização por electrospray (IES-EM/EM) foi desenvolvido para identificação dos nucleosídeos e bases em extratos de ascídias. Para este fim realizamos um estudo sistemático de preparo de amostra associado à análise de fragmentação de uma série de nucleosídeos e bases em IES-EM/EM. Estas abordagens constituem ferramentas analíticas importantes para screening rápido e determinação estrutural de nucleosídeos e bases em extratos de ascídias, os quais podem auxiliar nos estudos ecológicos do gênero.

Electrospray ionization tandem mass spectrometry (ESI-MS/MS) method was developed for identification of nucleosides and bases in extracts of ascidians. For this study a systematic sample preparation associated with the fragmentation pattern of a series of nucleosides and bases using ESI-MS/MS was done. These approaches constitute powerful analytical tools for rapid screening and structural assignment of nucleosides and bases in ascidian extracts.