Resumo Apresentamos um levantamento comentado de projetos financiados pelo programa BIOTA-Micro-organismos. Estes projetos geraram uma variada gama de resultados, incluindo a identificação de novos micro-organismos produtores de compostos antibacterianos, a caracterização de novas enzimas microbianas para usos industriais, classificação taxonômica de novos micro-organismos presentes em diversos ambientes, investigação de ecossistemas microbianos em solos e mangues e sua influência sobre plantas ameaçadas, e o sequenciamento de vários novos genomas microbianos derivados de metagenomas. Os resultados descritos demonstram o papel-chave de micro-organismos em ecossistemas importantes para atividades humanas, assim como o potencial que vários desses micro-organismos tem de contribuir para aplicações biotecnológicas cruciais para a sobrevivência humana no século 21.

Abstract We present a survey of projects that have been funded by FAPESP under the BIOTA-Microorganisms program. These projects generated a wide variety of results, including the identification of novel antibacterial-producing microorganisms, the characterization of novel microbial enzymes for industrial applications, taxonomic classification of novel microorganisms in several environments, investigation of the soil and mangrove microbial ecosystems and its influence on endangered plant species, and the sequencing of novel metagenome-assembled genomes. The results surveyed demonstrate the importance of microorganisms in environments that play important roles in human activities as well as the potential that many of these microorganisms have in contributing to biotechnological applications crucial for human survival in the 21st century.

MOLECULAR NETWORKS: AN ANALYSIS ON ANNOTATIONS AND DISCOVERY OF NEW ASSETS. To speed up the discovery of bioactive natural products (NP), chemists have sought advanced approaches in analytical and computational chemistry in attempt to organize and extract information from large data sets. In this sense, the molecular networks (MN) successfully organized enormous sets of mass spectrometry (MS) data together with samples metadata information in an intuitive visualization in the spectral similarity networks format. GNPS (Global Natural Products Social Molecular Networking), a free online platform for storing and processing MSn data, is a leading application of spectral matching with public databases aimed at the dereplication and discovery of new bioactive products through molecular networks. In this review, we address the concept of GNPS spectral similarity networks, as well as their complementary computational tools, benefits and limitations applied in NP studies associated with dereplication, chemical ecology, functional genetics and determination of biosynthetic pathways.

Metabolomics has played a central role in various areas of plant sciences, offering new perspectives for the advancement of agriculture, drug discovery, chemical ecology and taxonomy. Plant metabolomics (identification and quantification) aims to understand the relationship between biological systems and genetic, pathological and or environmental stimuli in terms of differential expression of the metabolism. Owing to the unique challenges, such studies require multidisciplinary skills involving biology, chemistry, statistics, and computer science for the extraction and complete understanding of information. In this sense, this review summarizes the main procedures that involve the steps of plant metabolomic study (design of experiments, sample preparation, analytical methods and data analysis), providing a comprehensive overview, showing the main challenges and limitations and possible solutions for the different approaches used.

Senna spectabilis (syn Cassia spectabilis) is one of the most important species within the Fabaceae family, natively found in Central and South America, as well as parts of Asia and Africa. Due to the extensive geographical distribution, this fast-growing tree produces a wide variety of bioactive secondary metabolites, being of special interest for chemical and pharmacological studies. Phytochemical investigations have shown that S. spectabilis produces over 40 constituents from different biosynthetic pathways, including piperidine alkaloids, pentacyclic terpenoids and anthraquinones, displaying antiproliferative, antitumoral and antifungal activities. Moreover, studies have also been conducted to identify endophytic and rizhospheric microorganisms associated to S. spectabilis and their chemical composition, enabling further elucidation of cadinane sesquiterpenoids, cytochalasins, depsipeptides and dibenzopirones. This review aims to provide an updated summary of the main features of S. spectabilis, compiling all currently available information on the chemical and pharmacological composition of its parts and its associated microorganisms.

The search for new sources of natural products steadily increased the use of bioinformatic tools that enabled efficient analysis of complex matrices. In this context, dereplication methods emerged as a fast way of identifying known compounds, accelerating the identification of bioactive chemotypes. Although 1H NMR is widely used as an analytical technique, few studies have been reported using it as a dereplication tool, primarily because of the spectral complexity. This work aims to create a new computational method that analyses 1H NMR data from Fusarium solani and F. oxysporum isolated from Senna spectabilis's rhizosphere through principal component analysis (PCA). The algorithm uses loading values to select important peaks that distinguish both species in PCA, allowing compound dereplication, even in highly similar profiles. As a result, the method, associated with other NMR experiments and information from an in-house Fusarium's metabolite library was able to distinguish different mycotoxins produced by both fungi, identifying fusaric acid and beauvericin for F. oxysporum and the depsipeptide HA23 from F. solani.

Um método racional e seletivo de cromatografia de alta eficiência acoplado à espectrometria de massas (CLAE-EM) com ionização por electrospray e analisador do tipo quadrupolo em tempo-de-voo em sequência (ESI-QToF-MS/MS) foi desenvolvido para a desreplicação de derivados fenólicos presentes em Qualea grandiflora e Qualea cordata. Os extratos para a análise foram selecionados por meio dos resultados de atividade antioxidante revelados no ensaio in vitro com DPPH. A análise por HPLC-ESI-QToF-MS/MS foi realizada por detecção contínua das relações massa/carga em alta resolução e pela técnica MS/MS de colisão induzida para fragmentação dos íons moleculares selecionados. A desreplicação da fração AcOEt do extrato hidroalcoólico dos caules de Q. grandiflora permitiu a detecção dos flavonoides: 3',4',5',5,6,7-hexahidroxi-8-metilflavanona, 8-metil-naringenina e 3',7-dimetoxi-8-metil-4',5,7-trihidroxiflavanona, assim como os derivativos da benzofenona (bis(4,6-dimetoxi-2-hidroxi-3-metilfenil)-metanona, 3',4'-dimetoxi-8-metil-5,6,7-trihidroxiflavanona, 7-metoxi-6-metil-3',4',5-trihidroxiflavanona, 6,8-dimetil-3'-metoxi-4',5,7-trihidroxiflavanona e 3',5'-dimetoxi-6,8-dimetil-4',5,7-trihidroxiflavanona) foram detectados na fração AcOEt do extrato hidroalcoólico das folhas de Q. cordata.

A rational and selective method using on-line high-performance liquid chromatography (HPLC) coupled with electrospray quadrupole time-of-flight tandem mass spectrometry (ESI-QToF-MS/MS) was established for the dereplication of phenolic derivatives from Qualea grandiflora and Qualea cordata. The selection of the extracts was based on the antioxidant capacity measured by in vitro DPPH assay. The HPLC-ESI-QToF-MS/MS analysis was conducted by on-flow detection, using high-resolution mass/ratio ions as well as collision induced MS/MS experiments for selected protonated ions. The dereplication of the EtOAc fraction from the hydro-alcohol extract from the stem bark of Q. grandiflora allowed the detection of the flavonoids: 3',4',5',5,6,7-hexahydroxy-8-methylflavanone, 8-methyl-naringenine and 3',7-dimethoxy-8-methyl-4',5,7-trihydroxyflavanone, as well as a benzophenone derivatives: bis(4,6-dimethoxy-2-hydroxy-3-methylphenyl)-metanone, 3',4'-dimethoxy-8-methyl-5,6,7-trihydroxyflavanone, 7-methoxy-6-methyl-3',4',5-trihydroxyflavanone, 6,8-dimethyl-3'-methoxy-4',5,7-trihydroxyflavanone and 3',5'-dimethoxy-6,8-dimethyl-4',5,7-trihydroxyflavanone were detected in the EtOAc fraction from the hydro-alcohol extract from the leaves of Q. cordata.

Brazilian biodiversity is a colossal source of secondary metabolites with remarkable structural features, which are valuable in further biodiscovery studies. In order to fully understand the relations and interactions of a living system with its surroundings, efforts in natural product chemistry are directed toward the challenge of detecting and identifying all the molecular components present in complex samples. It is plausible that this endeavor was born out of recent technological sophistication in secondary metabolite identification with sensitive spectroscopic instruments (MS and NMR) and higher resolving power of chromatographic systems, which allow a decrease in the amount of required sample and time to acquire data. Nevertheless, the escalation of data acquired in these analyses must be sorted with statistical and multi-way tools in order to select key information. Chromatography is also of paramount importance, more so when selected compounds need to be isolated for further investigation. However, in the course of pursuing a "greener" environment, new policies, with an aim to decrease the use of energy and solvents, are being developed and incorporated into analytical methods. Metabolomics could be an effective tool to answer questions on how living organisms in our huge biodiversity work and interact with their surroundings while also being strategic to the development of high value bio-derived products, such as phytotherapeutics and nutraceuticals. The incorporation of proper phytotherapeutics in the so-called Brazilian Unified Health System is considered an important factor for the urgent improvement and expansion of the Brazilian national health system. Furthermore, this approach could have a positive impact on the international interest toward scientific research developed in Brazil as well as the development of high value bio-derived products, which appear as an interesting economic opportunity in national and global markets. Thus, this study attempts to highlight the recent advances in analytical tools used in detection of secondary metabolites, which can be useful as bioproducts. It also emphasizes the potential avenues to be explored in Brazilian biodiversity, known for its rich chemical diversity.

The use of natural products has definitely been the most successful strategy in the discovery of novel medicines. Secondary metabolites from terrestrial and marine organisms have found considerable use in the treatment of numerous diseases and have been considered lead molecules both in their natural form and as templates for medicinal chemistry. This paper seeks to show the great value of secondary metabolites and emphasize the rich chemical diversity of Brazilian biodiversity. This natural chemical library remains understudied, but can be a useful source of new secondary metabolites with potential application as templates for drug discovery.

A utilização de parâmetros apenas morfológicos para posicionar taxonomicamente diversas espécies em sub-famílias e tribos na família Rubiaceae é bastante problemática devido à falta de informações sobre a distribuição geográfica e de características morfoanatômicas nos níveis hierárquicos mais baixos, como por exemplo, o gênero Chimarrhis. O perfil micromolecular de diferentes espécies pode auxiliar na delimitação de tribos indicando tendências filogenéticas mais completas entre as tribos das sub-famílias, já que os metabólitos secundários são expressões de adaptação, regulação e evolução de um determinado táxon. Nesse contexto, os alcalóides indólicos monoterpênicos isolados de Chimarrhis turbinata foram bastante úteis para embasar a classificação taxonômica feita por Robbrecht, em que posiciona Chimarrhis como um gênero da tribo Condamineae e subfamília Cinchonoideae.

The morphological parameters used to establish close connections among species taxonomically different into the Rubiaceae family is complex, mainly due to the lack of information on habitat and morphoanatomical characters in the lower hierarchic groups, for example, Chimarrhis genus. The micromolecular profile of delimited species into determined taxa can be useful to establish the boundaries among close taxonomic groups, and to indicate evolutionary phylogenetic trends into the taxa. Several indole alkaloids isolated from C. turbinata showed to be a valuable tool to support the taxonomic classification performed by Robbrecht, who established the most recent taxonomy for Rubiaceae, based on morphological characters, and concluded that Chimarrhis belong to Condamineae, and subfamily Cinchonoideae.

The phytochemical investigation of the flowers of Pterogyne nitens (Caesalpinioideae) resulted in the isolation and identification of nine phenolic derivatives, quercetin 3-O-sophoroside, taxifolin, astilbin, ourateacatechin, caffeic, ferulic, sinapic, chlorogenic and gallic acid, besides two guanidine alkaloids, pterogynine, pterogynidine. This is the first time these compounds have been reported in P. nitens flowers. As this is a monospecific genus, these secondary metabolites may have taxonomical significance. Their structures were assigned on the basis of spectroscopic analyses, including two-dimensional NMR techniques.

Chromatographic fractionation of the EtOH extract from the leaves of Swartzia langsdorffii afforded the pentacyclic triterpenes oleanolic acid and lupeol, and two saponins: oleanolic acid 3-sophoroside and the new ester 3-O-b-D-(6'-methyl)-glucopyranosyl-28-O-b-D-glucopyranosyl-oleanate. Their structures were elucidated from spectral data, including 2D NMR and HRESIMS experiments. Antifungal activity of all isolated compounds was evaluated, using phytopathogens Cladosporium cladosporioides and C. sphaerospermum, and human pathogens Candida albicans, C. krusei, C. parapsilosis and Cryptococcus neoformans.

Chromatographic fractionation of the organic extract from leaves of Ouratea multiflora afforded the flavone dimers heveaflavone, amentoflavone-7<FONT FACE=Symbol>¢¢</FONT>,4<FONT FACE=Symbol>¢¢¢</FONT>-dimethyl eter, podocarpusflavone-A and amentoflavone. Their structures were elucidated from spectral data, including 2D-NMR experiments of the natural substances. Biological activities of all isolates were evaluated, using antimicrobial assay against Gram-positive bacteria Staphylococcus aureus and Bacillus subtilis, cytotoxicity assay against mouse lymphoma (L5178) and KB cell lines, TLC screening for acetylcholinesterase inhibitors and antioxidant activity measured by DPPH test.

O fracionamento cromatográfico do extrato orgânico das folhas de Ouratea multiflora forneceu os flavonóides diméricos, heveaflavona, 7<FONT FACE=Symbol>¢¢</FONT>,4<FONT FACE=Symbol>¢¢¢</FONT>-dimetilamentoflavona, podocarpusflavona-A e amentoflavona. Suas estruturas foram elucidadas com base nos dados espectrais, incluindo experimentos bidimensionais de RMN, das substâncias naturais. A atividade antibiótica de todos os isolados foi avaliada, usando-se as bacterias Gram-positivas Staphylococcus aureus and Bacillus subtilis. Teste de citotoxicidade nas linhagens de linfoma de ratos (L5178) e KB também foram conduzidos para avaliar os extratos e os flavonóides isolados. a triagem biológica para a avaliação de atividade antioxidante e inibidora de acetil colinesterase foram conduzidas pela técnica da bioautografia com DPPH e teste pelo teste de Ellman respectivamente.

Várias hipóteses e constantes estudos são realizados com o intuito de elucidar o envelhecimento cutâneo. Para prevenir e contornar este processo torna-se necessário reforçar nossas defesas naturais antioxidantes endógenas. Diversas substâncias antioxidantes exógenas, como vitaminas, extratos vegetais, dentre outras, são utilizadas pela Cosmetologia nos produtos antienvelhecimento. O objetivo deste artigo é apresentar de que forma a Modelagem Molecular pode ser uma ferramenta útil na pesquisa por novas substâncias cosméticas antioxidantes para combater o envelhecimento cutâneo.

Some hypotheses and constants studies are made with intention to elucidate the aging process. To prevent and to attenuate the cutaneous aging it becomes necessary to strengthen our endogenous antioxidant natural defenses. Diverse exogenous antioxidant substances, as vitamins, vegetal extracts and others, have been used by the Cosmetology in antiaging products. The objective of this paper is to show how the Molecular Modeling can be an useful tool in the research for new antioxidant cosmetic substances to face the cutaneous aging.

Um novo biflavonol denominado chimarrosídeo (1) e oito flavonóis glicosilados adicionais (2-9) foram isolados das folhas de Chimarrhis turbinata. Suas estruturas foram elucidadas com base nos dados dos experimentos de RMN 1D e 2D, como: 3-O-rutinosilquercetina (2), 3-O-rutinosil-kaempferol (3), 3-O-galactopiranosil-(6->1)-ramnopiranosil kaempferol (4), 3-O-beta-galactopiranosil-(6->1)-alfa-ramnopiranosil-quercetina (5), 6-hidroxirutina (6), 3-O-galactopiranosil-kaempferol (7), 3-O-glucopiranosil-kaempferol (8) e 3-O-ramnopiranosil-(6->1)-glucopiranosil-(4 ->1)-ramnopiranosil-kaempferol (9). Adicionalmente, a catequina (10) e a procianidina B-3 (catequina-(4<FONT FACE=Symbol>a®</FONT>8)-catequina) (11) também foram isoladas. O extrato bruto, frações e compostos isolados foram avaliados quanto às suas propriedades antioxidantes no teste em CCD aspergida com solução de beta-caroteno, e teste espectrofotométrico utilizando o radical livre 1,1-difenil-2-picrilidrazila (DPPH). Os flavonóides 2, 5, 6, 10 e 11 apresentaram forte atividade antioxidante quando comparados com os padrões BHT e rutina.

A new biflavonol, named chimarrhoside (1), and eight known flavonol glycosides (2-9), were isolated from the leaves of Chimarrhis turbinata. Their structures were established on the basis of 1D and 2D NMR experiments as quercetin-3-O-rutinoside (2), kaempferol-3-O-rutinoside (3), kaempferol-3-O-alpha-L-rhamnopyranosyl-(1->6)- beta-D-galactopyranoside (4), quercetin-3-O-alpha-L-rhamnopyranosyl-(1->6)- beta-D-galactopyranoside (5), 6-hydroxy-rutin (6), kaempferol-3-O-D-galactopyranoside (7), kaempferol-3-O-D-glucopyranoside (8) and kaempferol-3-O-alpha-L-rhamnopyranosyl-(1->6)- alpha-L-rhamnopyranosyl-(1->4)-beta-D-glucopyranoside (9). In addition, catechin (10) and catechin-(4<FONT FACE=Symbol>a®</FONT>8)-catechin-procyanidin B-3) (11) were isolated. The crude extract, fractions and isolated compounds were evaluated for their antioxidative properties using an autographic assay based on beta-carotene bleaching on TLC plates, and spectrophotometric detection by reduction of the stable 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical. Flavonoids 2, 5, 6, 10 and 11 displayed strong free radical scavenging activity, when compared with the standards BHT and rutin.

O padrão de fragmentação de uma série homóloga de alcalóides piperidínicos isolados de Senna spectabilis foi investigado utilizando-se espectrometria de massas tandem com ionização por electrospray (ESI-EM/EM). A análise dos extratos EtOH e frações das flores e dos frutos de S. spectabilis por ESI-EM e ESI-EM/EM permitiu elucidar a estrutura de quatro componentes pertencentes a uma série homóloga destes alcalóides, bem como de quatro co-metabólitos inéditos. A elucidação estrutural dos co-metabólitos, com base no padrão de fragmentação de alcalóides piperidínicos previamente isolados, e confirmados pelos dados de EM de alta resolução, demonstra a importância desta técnica para a determinação do "perfil metabolômico" de uma espécie de importância farmacológica.

The fragmentation pattern of a homologous series of piperidine alkaloids isolated from S. spectabilis was investigated using electrospray ionization tandem mass spectrometry (ESI-MS/MS). The ESI-MS and ESI-MS/MS analyses of EtOH extracts and fractions from flowers and fruits of S. spectabilis allowed to elucidate the structures of four new compounds. The identification of these co-metabolites, based on the fragmentation patterns of previously isolated compounds, and further confirmed by accurate mass spectrometry defines this technique as a powerful tool to determine the "metabolomic profile" of species which has pharmacological importance.