Mônica T. Pupo

Endophytic Fungi: Natural Products, Enzymes and Biotransformation Reactions

Endophytes exhibit a complex web of interactions with host plants and with other endophytic microorganisms and therefore they have been intensively studied over the last several years as prolific sources of new and bioactive natural products. In fact, an impressive number of natural products have been produced by endophytic microorganisms. In addi- tion, some studies have shown endophytes to be good producers of useful enzymes to improve industrial processes. More recently, endophytes have also received attention as biocatalysts in the chemical transformation of natural products and drugs. Results have shown their ability to modify chemical structures with a high degree of stereospecificity. Some reac- tions are similar to those catalyzed by mammal phase I metabolism; therefore endophytes could be used as models for drug metabolism studies. This paper summarizes recent data on endophytes research as a source of novel and bioactive natural products, as producers of enzymes and their use on biotransformation processes.

Antibióticos: importância terapêutica e perspectivas para a descoberta e desenvolvimento de novos agentes

There is a continuous need for antibiotics, mainly with new mechanisms of action, since infectious diseases represent the second major cause of death in the world and bacteria resistance levels are high. This review describes the contribution of microbial natural products for the development of the major antibiotic classes, the mechanisms of action of current antibiotics, some modern approaches involving genetic tools for the discovery and development of new antibiotics from microbial products and antibiotics in clinical trials.

Global biogeographic sampling of bacterial secondary metabolism

Recent bacterial (meta)genome sequencing efforts suggest the existence of an enormous untapped reservoir of natural-product-encoding biosynthetic gene clusters in the environment. Here we use the pyro-sequencing of PCR amplicons derived from both nonribosomal peptide adenylation domains and polyketide ketosynthase domains to compare biosynthetic diversity in soil microbiomes from around the globe. We see large differences in domain populations from all except the most proximal and biome-similar samples, suggesting that most microbiomes will encode largely distinct collections of bacterial secondary metabolites. Our data indicate a correlation between two factors, geographic distance and biome-type, and the biosynthetic diversity found in soil environments. By assigning reads to known gene clusters we identify hotspots of biomedically relevant biosynthetic diversity. These observations not only provide new insights into the natural world, they also provide a road map for guiding future natural products discovery efforts. DOI: http://dx.doi.org/10.7554/eLife.05048.001

Diketopiperazines produced by an Aspergillus fumigatus Brazilian strain

Seven diketopiperazines, corresponding to the cyclos (L)-Pro-(L)-Phe, (L)-Pro-Gly, (L)-Pro-(L)-Pro, (L)-Pro-(L)-Val, (L)-4-OH-Pro-(L)-Leu, (L)-4-OH-Pro-(L)-Phe, and (L)-Pro-(L)-Leu, were isolated from the Aspergillus fumigatus fermentation broth. The relative and absolute stereochemistries were determined on the basis of NOESY experiments and by using a modified version of Marfeys method using HPLC, respectively.

Novel anthraquinone derivatives produced by Phoma sorghina, an endophyte found in association with the medicinal plant Tithonia diversifolia (Asteraceae)

Three known anthraquinones (1,7-dihydroxy-3-methyl-9,10-anthraquinone, 1,6-dihydroxy-3-methyl-9,10-anthraquinone and 1-hydroxy-3-methyl-9,10-anthraquinone), one new anthraquinone (1,7-dihydroxy-3-hydroxymethyl-9,10-anthraquinone), and two new hexahydroanthraquinone derivatives, dendryols E and F, were isolated from the culture of the endophytic fungus Phoma sorghina, found in association with Tithonia diversifolia (Asteraceae). Their structures were identified on the basis of spectroscopic data, mainly 1D and 2D NMR.

Endophytic fungi found in association with Smallanthus sonchifolius (Asteraceae) as resourceful producers of cytotoxic bioactive natural products

Smallanthus sonchifolius is a traditional Andean plant which has been cultured mainly in Brazil, Japan and New Zealand due to its medicinal properties. A study of the endophytic fungi associated to the plant was carried out in order to characterize new cytotoxic agents. Thirty two fungal strains were isolated and submitted to cultivation and extraction producing 186 extracts. Of these, 12% displayed moderate to high cytotoxic activities and were considered promising anticancer compound sources. The ethyl acetate fractions of Nigrospora sphaerica and Phoma betae liquid fermentations contained the synergistic compounds 8‐hydroxy‐6‐methoxy‐3‐methylisocoumarin and (22E,24R)‐ergosta‐4,6,8(14),22‐tetraen‐3‐one which are potential compounds for drug discovery. Another isolated compound, pimara‐7,15‐dien‐3‐β‐ol diterpene is being characterized for the first time through a detailed spectroscopic analysis including GC/MS, homo‐ and hetero‐nuclear correlated NMR experiments (HMQC, HMBC, COSY and NOEdiff) along with its optical rotation. (© 2009 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)

A Mixed Culture of Endophytic Fungi Increases Production of Antifungal Polyketides

Secondary metabolites produced by endophytic microorganisms can provide benefits to host plants, such as stimulating growth and enhancing the plant’s resistance toward biotic and abiotic factors. During its life, a host plant may be inhabited by many species of endophytes within a restrictive environment. This condition can stimulate secondary metabolite production that improves microbial competition and may consequently affect both the neighboring microorganisms and the host plant. The interactions between the endophytes that co-habit the same host plant have been studied. However, the effect of these interactions on the host plant has remained neglected. When using mixed microbial cultures, we found that the endophytic fungus Alternaria tenuissima significantly increased the production of some polyketides, including antifungal stemphyperylenol in response to the endophytic Nigrospora sphaerica. Biological activity assays revealed that stemphyperylenol can cause cytotoxic effects against N. sphaerica, although no phytotoxicity was observed in the host plant Smallanthus sonchifolius, even at concentrations much higher than those toxic to the fungus. The polyketides produced by A. tenuissima may be important for the ecological relationships between endophyte-endophyte and endophytes-host plants in the natural environment.

Diketopiperazines produced by endophytic fungi found in association with two Asteraceae species.

Diketopiperazine (DKP) derivatives, named colletopiperazine, fusaperazine C and E as well as four known DKPs were isolated from cultures of Colletotrichum gloeosporioides, Penicillium crustosum, both endophytic fungi isolated from Viguiera robusta, and a Fusarium spp., an endophyte of Viguiera arenaria, respectively. Their structures were established on the basis of their spectroscopic data. Conformational analysis of two known DKPs showed that folded conformations were as energetically stable as the extended one.

Strategies for the isolation and identification of trypanocidal compounds from the Rutales

Crude extracts of Rutales species were tested in vitro against the trypomastigote form of Trypanosoma cruzi at 4 mg/mL, and 20% of them showed significant activity (80%). Their inhibitory activity against the glycolytic enzyme GAPDH from T. cruzi has also been evaluated at the concentrations of 100 and 200 mg/mL. Additionally, the inhibitory activity of 13 purified coumarins were also assayed against T. cruzi-GAPDH. Chalepin was the most active substance with IC50 = 64 mM. The 3D structure of the complex chalepin-enzyme was elucidated by X-ray crystallography, revealing the architecture of the interactions between the inhibitor and the enzyme active site.

INTRODUÇÃO A MODELAGEM MOLECULAR DE FÁRMACOS NO CURSO EXPERIMENTAL DE QUÍMICA FARMACÊUTICA

Molecular Modeling is an important tool in drug design and it is very useful to predict biological activity from a library of compounds. A wide variety of computer programs and methods have been developed to visualize the tridimensional geometry and calculate physical properties of drugs. In this work, we describe a practical approach of molecular modeling as a powerful tool to study structure-activity relationships of drugs, including some antibacterials, hormones, cholinergic and adrenergic agents. At first, the students learn how to draw 3D structures and use them to perform conformational and molecular analysis. Thus, they compare drugs with similar pharmacological activity by superimposing one structure on the top of another and evaluate the geometry and physical properties.