Barcode e bioprospecção de metabólitos das algas marinhas Laurencia aldingensis, L. dendroidea e Laurenciella sp. (Ceramiales, Rhodophyta) / Barcode and bioprospecting of metabolites from the marine algae Laurencia aldingensis, L. dendroidea e Laurenciella sp. (Ceramiales, Rhodophyta)

A natureza e diversidade das estruturas químicas com atividade farmacológica que se tem encontrado nos organismos marinhos justificam a busca por novos compostos que são de interesse nas mais diversas áreas de aplicação. As espécies de macroalgas vermelhas, em especial Laurencia spp., merecem destaque pela enorme variedade de terpenos e acetogeninas que produzem, sendo consideradas de grande potencial na produção de novos fármacos. O estudo de seus constituintes pode fornecer importantes subsídios para a quimiotaxonomia, ecologia química, caracterização das espécies e avaliação do potencial biotecnológico. Baseado nisso, Laurencia aldingensis, L. dendroidea e Laurenciella sp. foram selecionadas para o presente estudo para isolamento, caracterização e teste de atividades biológicas dos seus compostos. A técnica do DNA barcoding foi utilizada como ferramenta de diagnóstico para garantir a similaridade entre as amostras de cada espécie, que foram coletadas em época e locais diferentes. Do extrato orgânico de Laurencia aldingensis, nove substâncias foram isoladas, sendo quatro esfingosinas (1-4), três terpenos (5-7) e duas novas substâncias halogenadas (8 e 9). Do extrato orgânico de Laurencia dendroidea formam isolados dois terpenos halogenados conhecidos (10, 11) e, do extrato de Laurenciella sp. três novas substâncias halogenadas alifáticas insaturadas (12-14), assim como um ácido graxo (15) e um esterol (16) conhecidos. Dentre elas, a 8 apresentou atividade citotóxica, mas não se mostrou seletivo, e as substâncias 4 e 11 apresentaram atividade esquistossomicida, bastante promissora. No entanto, nenhum deles apresentou atividade antioxidante. Diante desta investigação, podemos dizer que as informações geradas com os estudos de Laurencia aldingensis, L. dendroidea e Laurenciella sp. expandiram significantemente o conhecimento no que tange a diversidade química no gênero e o potencial biológico-farmacêutico dos mesmos

The nature and diversity of chemical structures with pharmacological activity that have been found in marine organisms justifies the search for new compounds that may have applications in various areas of interest. Species of red seaweeds, especially Laurencia spp., are special because of the unprecedented variety of terpenes and acetogenins they produce that are considered potentially useful for the production of new drugs. Study of their constituents can also provide important insights relating to their chemotaxonomy, chemical ecology, characterization of species and biotechnological potential. On this basis Laurencia aldingensis, L. dendroidea and Laurenciella sp., were selected for study and isolation, characterization, and biological activity assessment of isolatable quantities of their compounds. The technique of DNA barcoding was used as a diagnostic tool to ensure similarity between samples of each species collected at different times and places. From the organic extract of Laurencia aldingensis nine compounds were isolated; four sphingosines (1-4), three terpenes (5-7) and other two new halogenated compound (8, 9). From the organic extract of Laurencia dendroidea two known halogenated terpenes (10, 11) were isolated while from a similar extract of Laurenciella sp., three new halogenated aliphatic compounds (12-14) were isolated together with known fatty acid (15) and sterol (16). Among all isolates, 8 demonstrated unspecific cytotoxic activity and compounds 4 and 11 showed promising schistosomicidal activity. In applied antioxidant assays none of the isolates we noted to have activity. From the overall investigation it is also clear that the information gleaned from the studies of Laurencia aldingensis, L. dendroidea and Laurenciella sp., significantly expanded our knowledge base concerning chemical diversity in the genus Laurencia and their biological-pharmaceutical potential

Prospecção de bioativos farmacológicos em algas marinhas Rhodophyta e Heterokontophyta e avaliação de citotoxicidade / Prospection of pharmaceutical bioactive compounds in marine algae Rhodophyta and Heterokontophyta and cytotoxicity evaluation

Muitas drogas terapêuticas produzidas pela indústria farmacêutica são estruturas químicas isoladas de organismos encontrados na natureza ou moléculas baseadas nelas. Podem ser incluídas nesse grupo drogas isoladas de organismos marinhos, como corais, esponjas e algas marinhas, conhecidos como produtores de grandes quantidades de metabólitos secundários. Com base neste fato o presente estudo teve como objetivo realizar a prospecção de moléculas bioativas com propósito farmacológico, em extratos de algas marinhas vermelhas (Rhodophyta) e pardas (Heterokontophyta) coletadas no litoral brasileiro. A prospecção foi realizada por meio de avaliação de seus potenciais antioxidante, antibacteriano, antifúngico, anticancerígeno, e antiparasitário contra organismos causadores de leishmaniose e esquistossomose. Para as avaliações foram empregadas os extratos supercríticos de 5 espécies diferentes, sendo 2 pardas: Dictyota dichotoma e D. menstrualis e três vermelhas: Chondria littoralis, Spyridia hypnoides e Plocamium brasiliense. Os extratos foram avaliados quanto aos seus potenciais bioativos e os resultados mais promissores foram selecionados para as etapas seguintes do fracionamento. Em uma avaliação geral os extratos apresentaram bons resultados e representam uma potencial fonte de bioativos. Os extratos das espécies de D. dichotoma e D. menstrualis foram então submetidos a um procedimento de fracionamento bioguiado pela atividade esquistossomicida. Incorporou-se ainda um terceiro extrato de D. mertensii aos estudos e todas as etapas do fracionamento foram monitoradas por LC-MS. Comparando-se as massas detectadas em todas as frações que apresentaram atividade, para os 3 extratos, foi verificado que a substância de m/z 271,24 estava presente em todas elas, portanto os procedimentos de isolamento foram direcionados a esta molécula para a qual foi possível isolar 7 mg. Diferentemente do que era esperado a molécula quando avaliada isoladamente não apresentou atividade esquistossomicida, levando a hipótese de que a atividade seja decorrente de uma molécula diferente para cada espécie ou ainda que a mesma seja decorrente de uma interação com outras substâncias por um mecanismo de ação aditivo ou sinérgico. O trabalho avaliado de forma geral apresentou resultados promissores e representa um grande embasamento para servir como base para posteriores trabalho de fracionamento

Several therapeutic drugs manufactured by the pharmaceutical industry are chemical structures isolated from organisms that are found in nature or molecules based on that. May be included at this group drugs isolated from marine organisms, like corals, sponges and seaweeds, known as great secondary metabolites producers. Based on this facts the objective of the present study is to perform a prospection study to achieve bioactive molecules with pharmaceutical purposes, on extracts made from red (Rhodophyta) and brown (Heterokontophyta) seaweed collected in the Brazilian shore. The prospection studies was performed by means of evaluation of the antioxidant, antibacterial, antifungal, anticancer and antiparasitic (against Leishmania and Schistosoma) potential. In the evaluation were tested the supercritical extracts of 5 different species, including 2 brown species: Dictyota dichotoma and D. menstrualis and 3 red species: Chondria littoralis, Spyridia hypnoides and Plocamium brasiliense. The extracts were evaluated by their potential bioactive compounds and the most promising results were selected for the following fractionation steps. Overall the extracts have shown good results and may be represent a potential source of bioactive molecules. The extracts of both D. dichotoma and D. menstrualis were submitted to a bioguided fractionation process by their antischistosomal activities. It was still included a third extract from D. mertensii to the studies and every step was monitored by LC-MS techniques. Comparing the detected mass for each active fraction, it was observed the presence of a substance with m/z 271,24 in all of the extracts, so the isolating procedures were directed to obtain that specific molecule, which was obtained in a biomass of 7 mg. Differently than expected the molecule when evaluated isolated do not show the antischistosomal activity, leading to the hypothesis that the activity was related to different molecules for each species or even the observed effect is resulted by an interaction mechanism with another substances by an additive or synergist mechanism. The overall evaluation of the whole work show some promising results and it represent a great support for future fractionation works

Efecto de la asociación alga-bacteria Bostrychia calliptera (Rhodomelaceae) en el porcentaje de remoción de cromo en laboratorio / The effect of chromium removal by Algae-bacteria Bostrychia calliptera (Rhodomelaceae) consortia under laboratory conditions

Water pollution is one of the most important environmental problems worldwide. Recently, biotechnology studies have oriented efforts to study algae-bacterium consortia with the aim to understand the mechanisms to find a possible solution in environmental sciences. This study determined the percentage of chromium removal by the alga-bacterium association exposed to a set of different chromium concentrations under controlled in vitro conditions. Wild plants of Bostrychia calliptera associated with bacterial populations were collected from Dagua River, Pacific coast of Colombia, and were monitored in the laboratory. The trial was conducted with synthetic seawater in bioreactors at two chromium levels: 5 and 10mg/L, and four different experimental treatments: i) algae-bacteria (AB), ii) algae with antibiotic (AA), iii) algal surface sediment, Natural Bacterial Consortium (CBN), and iv) the control without algae or bacteria. The experimental design followed a model of two factors (chromium concentration x combination types) with repeated measures using one factor. The microbial population behavior and the chromium concentration percentage were monitored by using atomic absorption spectroscopy (AAS). According to the data, Algae-bacteria (AB) treatment was the most efficient combination at 10mg/L (87%), whereas the bacterial consortia (CBN) was the most efficient at 5mg/L (62.85%). The results showed significant differences of chromium uptake between algae-bacteria (AB) and natural bacterial consortia (CBN), meaning the importance of those treatments in the chromium removal from coastal waters.

Para determinar el porcentaje de remoción de cromo en la asociación alga-bacteria, se tomaron ejemplares del alga Bostrychia calliptera de pneumatóforos de Avicennia germinans y Rizophora mangle, en la desembocadura del Río Dagua, Pacífico colombiano. El ensayo se realizó in vitro en agua marina sintética a dos concentraciones de cromo 5 y 10mg/L, empleando biorreactores con cuatro tratamientos; i) material Alga-Bacteria (AB), ii) material algal con antibiótico Alga-antibiótico (AA), iii) consorcio bacteriano natural (CBN) y iv) control sin presencia de B. calliptera ni bacteria. Se monitoreó el comportamiento de poblaciones bacterianas y el porcentaje de disminución de cromo mediante AAS (Espectroscopía de absorción atómica). Se obtuvo diferencias significativas entre los tratamientos Alga-bacteria (AB) y el consorcio bacteriano natural (CBN), siendo Alga-bacteria más eficiente a 10mg/L (87%). El consorcio bacteriano natural CBN obtuvo mayor porcentaje de remoción a bajas concentraciones de cromo a 5ppm (62.85%). Los resultados mostraron una interacción positiva entre las bacterias asociadas (CBN) a la superficie del alga roja B. calliptera en su proceso acumulativo de cromo.

A simple method for efficient extraction and purification of C-phycocyanin from Spirulina platensis Geitler.

Phycocyanin is a major light harvesting accessory pigment of red algae and cyanobacteria. In the light of its many commercial applications in food and pharmaceutical industry, purity of the pigment plays a major role. Pharmaceutical industry demands a highly pure phycocyanin with A620/280 ratio of 4 and food industry a ratio of 2. In the present study phycocyanin was extracted in sodium phosphate buffer (pH 7) after macerating in liquid nitrogen. The crude phycocyanin thus extracted was precipitated with 50% ammonium sulphate, purified by dialysis and finally by gel filtration chromatography. Pure phycocyanin was finally obtained with an A620/A280 value of 4.98.