RESUMO
Although many advances have been achieved to treat aggressive tumours, cancer remains a leading cause of death and a public health problem worldwide. Among the main approaches for the discovery of new bioactive agents, the prospect of microbial secondary metabolites represents an effective source for the development of drug leads. In this study, we investigated the actinobacterial diversity associated with an endemic Antarctic species, Deschampsia antarctica, by integrated culture-dependent and culture-independent methods and acknowledged this niche as a reservoir of bioactive strains for the production of antitumour compounds. The 16S rRNA-based analysis showed the predominance of the Actinomycetales order, a well-known group of bioactive metabolite producers belonging to the Actinobacteria phylum. Cultivation techniques were applied, and 72 psychrotolerant Actinobacteria strains belonging to the genera Actinoplanes, Arthrobacter, Kribbella, Mycobacterium, Nocardia, Pilimelia, Pseudarthrobacter, Rhodococcus, Streptacidiphilus, Streptomyces and Tsukamurella were identified. The secondary metabolites were screened, and 17 isolates were identified as promising antitumour compound producers. However, the bio-guided assay showed a pronounced antiproliferative activity for the crude extracts of Streptomyces sp. CMAA 1527 and Streptomyces sp. CMAA 1653. The TGI and LC50 values revealed the potential of these natural products to control the proliferation of breast (MCF-7), glioblastoma (U251), lung/non-small (NCI-H460) and kidney (786-0) human cancer cell lines. Cinerubin B and actinomycin V were the predominant compounds identified in Streptomyces sp. CMAA 1527 and Streptomyces sp. CMAA 1653, respectively. Our results suggest that the rhizosphere of D. antarctica represents a prominent reservoir of bioactive actinobacteria strains and reveals it as an important environment for potential antitumour agents.
Assuntos
Actinobacteria , Técnicas de Cultura/métodos , Descoberta de Drogas , Neoplasias/patologia , Actinobacteria/metabolismo , Actinomycetales/metabolismo , Regiões Antárticas , Antraciclinas/isolamento & purificação , Antraciclinas/metabolismo , Antraciclinas/farmacologia , Antineoplásicos/isolamento & purificação , Antineoplásicos/metabolismo , Antineoplásicos/farmacologia , Fatores Biológicos/biossíntese , Fatores Biológicos/isolamento & purificação , Fatores Biológicos/farmacologia , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Dactinomicina/biossíntese , Dactinomicina/isolamento & purificação , Dactinomicina/farmacologia , Humanos , Streptomyces/metabolismoRESUMO
A new polycyclic antibiotic, pradimicin-IRD, was isolated from actinobacteria Amycolatopsis sp. IRD-009 recovered from soil of Brazilian rainforest undergoing restoration area. This molecule is the major compound produced in solid culture media. The new compound was detected by a focused method of precursor ion (high-performance liquid chromatography coupled to tandem mass spectrometer) developed previously to identify unusual aminoglycosyl sugar moieties. The compound was isolated and its structure was, therefore, elucidated by high-resolution mass spectrometry, and 1D and 2D nuclear magnetic resonance experiments. Pradimicin-IRD displayed potential antimicrobial activity against Streptococcus agalactiae (MIC 3.1 µg/mL), Pseudomonas aeruginosa (MIC 3.1 µg/mL) and Staphylococcus aureus (MIC 3.1 µg/mL), and also cytotoxicity against tumour and non-tumour cell lines with IC50 values ranging from 0.8 µM in HCT-116 colon carcinoma cells to 2.7 µM in MM 200 melanoma cells. Particularly, these biological properties are described for the first time for this chemical class.
Assuntos
Actinobacteria/química , Antraciclinas/isolamento & purificação , Antibacterianos/isolamento & purificação , Antraciclinas/química , Antibacterianos/química , Brasil , Linhagem Celular , Linhagem Celular Tumoral , Cromatografia Líquida de Alta Pressão , Humanos , Concentração Inibidora 50 , Testes de Sensibilidade Microbiana , Pseudomonas aeruginosa/efeitos dos fármacos , Microbiologia do Solo , Staphylococcus aureus/efeitos dos fármacos , Espectrometria de Massas em TandemRESUMO
The search for new compounds effective against Mycobacterium tuberculosis is still a priority in medicine. The evaluation of microorganisms isolated from non-conventional locations offers an alternative to look for new compounds with antimicrobial activity. Endophytes have been successfully explored as source of bioactive compounds. In the present work we studied the nature and antimycobacterial activity of a compound produced by Streptomyces scabrisporus, an endophyte isolated from the medicinal plant Amphipterygium adstringens. The active compound was detected as the main secondary metabolite present in organic extracts of the streptomycete and identified by NMR spectroscopic data as steffimycin B (StefB). This anthracycline displayed a good activity against M. tuberculosis H37Rv ATCC 27294 strain, with MIC100 and SI values of 7.8 µg/mL and 6.42, respectively. When tested against the rifampin mono resistant M. tuberculosis Mtb-209 pathogen strain, a better activity was observed (MIC100 of 3.9 µg/mL), suggesting a different action mechanism of StefB from that of rifampin. Our results supported the endophyte Streptomyces scabrisporus as a good source of StefB for tuberculosis treatment, as this anthracycline displayed a strong bactericidal effect against M. tuberculosis, one of the oldest and more dangerous human pathogens causing human mortality.
Assuntos
Antraciclinas/farmacologia , Sapindaceae/metabolismo , Anacardiaceae , Antraciclinas/isolamento & purificação , Antraciclinas/metabolismo , Anti-Infecciosos/farmacologia , Antituberculosos , Endófitos/isolamento & purificação , Testes de Sensibilidade Microbiana , Mycobacterium tuberculosis/efeitos dos fármacos , Extratos Vegetais/isolamento & purificação , Plantas Medicinais/metabolismo , Sapindaceae/toxicidade , Streptomycetaceae/metabolismoRESUMO
Four new anthracyclinones, 4,6,11-trihydroxy-9-propyltetracene-5,12-dione (1), 1-methoxy-9-propyltetracene-6,11-dione (2), 7,8,9,10-tetrahydro-9-hydroxy-1-methoxy-9-propyltetracene-6,11-dione (3), and 10ß-carbomethoxy-7,8,9,10-tetrahydro-4,6,7α,9α,11-pentahydroxy-9-propyltetracene-5,12-dione (4), were isolated from a strain of Micromonospora sp. associated with the tunicate Eudistoma vannamei. All structures were established by 1D and 2D NMR (COSY, HSQC, HMBC, NOESY) and HRESIMS experiments. Compounds 1 and 4 were cytotoxic against the HCT-8 human colon adenocarcinoma cell line, with IC(50) values of 12.7 and 6.2 µM, respectively, while compounds 2 and 3 were inactive.
Assuntos
Antraciclinas/isolamento & purificação , Antraquinonas/isolamento & purificação , Antineoplásicos/isolamento & purificação , Micromonospora/química , Animais , Antraciclinas/química , Antraciclinas/farmacologia , Antraquinonas/química , Antraquinonas/farmacologia , Antineoplásicos/química , Antineoplásicos/farmacologia , Ensaios de Seleção de Medicamentos Antitumorais , Humanos , Estrutura Molecular , Ressonância Magnética Nuclear Biomolecular , UrocordadosRESUMO
Aqueous two-phase systems (ATPS) were applied for extraction of small molecules (polycetides) - retamycin, an anthracyclin, and two red pigments, rubropunctamin and monascorubramin - from the whole culture media of Streptomyces olindensis and Monascus purpureus. ATPS allows, in one step, the separation of the small hydrophobic molecules in the PEG rich phase, from the filamentous microorganisms, which remains in the salt phase. Through experimental designs, the main variables and their levels were defined, as follows: for retamycin extraction, PEG 6000 (10%, w/w), phosphate at 20% (w/w) and pH 6.0 led to the higher partition coefficient, K(r) = 8.2, and yield = 91.3%; for red pigments, the statistical analysis indicate PEG 6000 (20%, w/w) and phosphate at 15% (w/w), for a high partition coefficient, (K(pig) = 113 and 150).
Assuntos
Antraciclinas/isolamento & purificação , Cromatografia Líquida/métodos , Monascus/química , Naftacenos/química , Naftacenos/isolamento & purificação , Streptomyces/química , Trissacarídeos/química , Antraciclinas/química , Configuração de Carboidratos , Meios de Cultura , Dados de Sequência MolecularRESUMO
Fed-batch runs were performed in order to correlate the production of retamycin, an anthracycline antibiotic produced by Streptomyces olindensis in submerged cultures, with the specific growth rate. Maximum retamycin production was achieved with an exponential feed rate, controlling the specific growth rate at a low value (0.03 h-1, about 10% of the maximum specific growth rate). Control of the specific growth rate at higher values (0.10 and 0.17 h-1) caused a decrease in antibiotic production. Morphology, assessed by image analysis, was shown to be highly relevant in this process. Cell growth mainly in the form of clumps (90% clumps and 10% free filaments) led to better results than growth as clumps (75%) and free filaments (25%).
Assuntos
Antraciclinas , Streptomyces/metabolismo , Antraciclinas/isolamento & purificação , Antraciclinas/metabolismo , Antineoplásicos/isolamento & purificação , Antineoplásicos/metabolismo , Biomassa , Reatores Biológicos/microbiologia , Meios de Cultura , Fermentação , Glucose/metabolismo , Streptomyces/citologia , Streptomyces/crescimento & desenvolvimentoRESUMO
Cosmomycin D (CosD) is the major constituent fraction isolated from a culture of Streptomyces olindensis ICB20. The ability of this compound to intercalate with double-stranded DNA was studied by gel mobility shift assays and electrospray ionization mass spectrometry (ESI-MS). ESI-MS experiments showed that the complex of CosD with 16-mer double-stranded DNA was at least as stable as a complex of daunorubicin with the same DNA sequence. This is the first study showing DNA binding properties of an anthracycline containing a beta-rhodomycinone aglycone chromophore O-linked to two trisaccharide chains.