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Arq. bras. med. vet. zootec. (Online) ; 71(6): 1993-1998, Nov.-Dec. 2019. tab, graf
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-1055109


The effect of three Streptomyces strains (N7, RL8 and V4) and a mixture of Bacillus (BMix) on the growth (Weight, Size) and superoxide dismutase activity (SOD) in hatchery-reared juvenile oysters Crassostrea corteziensis and Crassostrea sikamea was investigated to determine their probiotic potential. Microorganisms were added to culture water at 1×106 CFU/ml once a day during 30 days and all oysters fed daily a microalgae mix. Juveniles of C. sikamea treated with strains N7, RL8 and V4 had a significant weight gain compared to the control group. C. corteziensis juveniles treated with strains RL8 and BMix showed a significantly higher weight gain than the control group. No significant size increase was observed in any treated group for both oyster species. SOD activity significantly increased in C. sikamea treated with RL8 and with RL8, N7 and BMix in C. corteziensis. Streptomyces strains RL8 and N7 emerge as promising probiotic agents to cultivate C. sikamea and C. corteziensis and may also be useful to other molluscs and marine invertebrates .(AU)

O efeito de três culturas Streptomyces (N7, RL8 e V4) e uma mistura de Bacillus (BMix) sobre o cresimento (Peso, Tamanho) e atividade superóxido dismutase (SOD) em ostras jovens Crassostrea corteziensis e Crassostrea sikamea cultivadas artificalmente foi investigado para determinar seu potencial probiótico. Microorganismos foram adicionados à água de cultura a 1×10 6 CFU/ml uma vez por dia durante 30 dias e todas as ostras foram alimentadas diariamente com uma mistura de microalgas. Jovens C. sikamea tratados com culturas N7, RL8 e V4 tiveram ganho de peso significativo quando comparado ao grupo de controle. Jovens C. corteziensis tratados com culturas RL8 e BMix demonstraram peso significativamente mais algo que o grupo de controle. Nenhum aumento em tamanho foi observado em grupos tratados em ambas espécies. A atividade SOD foi significamente aumentada em C. sikamea treatado com RL8 e com RL8, N7 e BMix em C. corteziensis. Culturas Streptomyces RL8 e N7 surgem como agentes probióticos promissores para o cultivo de C. sikamea e C. corteziensis e podem ser úteis para outros moluscos animais marinhos invertebrados.(AU)

Animais , Streptomyces , Superóxido Dismutase/análise , Probióticos/administração & dosagem , Crassostrea/crescimento & desenvolvimento
Biosci. j. (Online) ; 35(3): 910-919, may./jun. 2019. tab, graf
Artigo em Inglês | LILACS | ID: biblio-1048701


To improve kitasamycin biosynthesis by Streptomyces kitasatoensis Z-7, the addition of two precursors, sodium acetate and ethyl acetate, to the fermentation medium was evaluated. Ethyl acetate was the most effective precursor compared with control conditions; In a 15-L fermentor, the kitasamycin titer was 21% higher when 0.48% ethyl acetate was added compared to control conditions. Content of the A5 component increased by 5.1%, and the A4 content decreased slightly compared to that of the control. During kitasamycin synthesis, intracellular and extracellular concentrations of acetic acid were higher for S. kitasatoensis Z-7 supplemented with ethyl acetate than for the non-supplemented strain, and the activities of acyl-CoA synthetases, acyl-phosphotransferases, and acyl-kinases were also significantly increased, suggesting that increased acetyl-CoA levels can explain the high kitasamycin titer. These findings may improve the industrial-scale production of kitasamycin for clinical use, and the addition of 0.48% ethyl acetate as precursors in the medium at the beginning of cultivation was a new method to mitigate the negative influence on the cell growth of excess precursor.

Para melhorar a biossíntese de kitasamicina por Streptomyces kitasatoensis Z-7, a adição de dois precursores, acetato de sódio e acetato de etila, ao meio de fermentação foi avaliada. O acetato de etila foi o precursor mais efetivo em comparação com as condições de controle; Em um fermentador de 15 L, o título de kitasamicina foi 21% maior quando 0,48% de acetato de etila foi adicionado em comparação com as condições de controle. O conteúdo do componente A5 aumentou 5,1%, e o conteúdo A4 diminuiu ligeiramente em comparação com o do controle. Durante a síntese de kitasamicina, as concentrações intracelulares e extracelulares de ácido acético foram maiores para S. kitasatoensis Z-7 suplementado com acetato de etila do que para a cepa não suplementada, e as atividades de acil-CoA sintetases, acil-fosfotransferases e acil-cinases também foram significativamente aumentadas, sugerindo que níveis aumentados de acetil-CoA podem explicar o alto título de kitasamicina. Esses achados podem melhorar a produção em escala industrial da kitasamicina para uso clínico, e a adição de 0,48% de acetato de etila como precursores no meio no início do cultivo foi um novo método para mitigar a influência negativa no crescimento celular do excesso de precursor.

Streptomyces , Kitasamicina , Fermentação , Antibacterianos
Braz. j. microbiol ; 49(4): 816-822, Oct.-Dec. 2018. tab, graf
Artigo em Inglês | LILACS | ID: biblio-974289


ABSTRACT Fifty seven soil-borne actinomycete strains were assessed for the antibiotic production. Two of the most active isolates, designed as Streptomyces ST-13 and DK-15 exhibited a broad range of antimicrobial activity and therefore they were selected for HPLC fractionation against the most suppressed bacteria Staphylococcus aureus (ST-13) and Chromobacterium violaceum (DK-15). LC/MS analysis of extracts showed the presence of polyketides factumycin (DK15) and tetrangomycin (ST13). The taxonomic position of the antibiotic-producing actinomycetes was determined using a polyphasic approach. Phenotypic characterization and 16S rRNA gene sequence analysis of the isolates matched those described for members of the genus Streptomyces. DK-15 strain exhibited the highest 16S rRNA gene sequence similarity to Streptomyces globosus DSM-40815 (T) and Streptomyces toxytricini DSM-40178 (T) and ST-13 strain to Streptomyces ederensis DSM-40741 (T) and Streptomyces phaeochromogenes DSM-40073 (T). For the proper identification, MALDI-TOF/MS profile of whole-cell proteins led to the identification of S. globosus DK-15 (accession number: KX527570) and S. ederensis ST13 (accession number: KX527568). To our knowledge, there is no report about the production of these antibiotics by S.globosus and S. ederensis, thus isolates DK15 and ST13 identified as S. globosus DK-15 and S.ederensis ST-13 can be considered as new sources of these unique antibacterial metabolites.

Streptomyces/isolamento & purificação , Streptomyces/metabolismo , Antibacterianos/biossíntese , Filogenia , Piridonas/metabolismo , Microbiologia do Solo , Streptomyces/classificação , Streptomyces/genética , Benzo(a)Antracenos/metabolismo , DNA Bacteriano/genética , Técnicas de Tipagem Bacteriana
Braz. j. microbiol ; 49(4): 731-741, Oct.-Dec. 2018. tab, graf
Artigo em Inglês | LILACS | ID: biblio-974291


ABSTRACT A bacterium isolated from Sterkfontein dam was confirmed to produce bioflocculant with excellent flocculation activity. The 16S rDNA nucleotide sequence analyses revealed the bacteria to have 99% similarity to Streptomyces platensis strain HBUM174787 and the sequence was deposited in the Genbank as Streptomyces platensis with accession number FJ 486385.1. Culture conditions for optimal production of the bioflocculant included glucose as a sole carbon source, resulting in flocculating activity of 90%. Other optimal conditions included: peptone as nitrogen source; presence of Mg2+ as cations and inoculum size of 1.0% (v/v) at neutral pH of 7. Optimum dose of the purified bioflocculant for the clarification of 4 g/L kaolin clay suspension at neutral pH was 0.2 mg/mL. Energy Dispersive X-ray analysis confirmed elemental composition of the purified bioflocculant in mass proportion (%w/w): carbon (21.41), oxygen (35.59), sulphur (26.16), nitrogen (0.62) and potassium (7.48). Fourier Transform Infrared Spectroscopy (FTIR) indicated the presence of hydroxyl, carboxyl, methoxyl and amino group in the bioflocculant. The bioflocculant produced by S. platensis removed chemical oxygen demand (COD) in river water and meat processing wastewater at efficiencies of 63.1 and 46.6% respectively and reduced their turbidity by 84.3 and 75.6% respectively. The high flocculating rate and removal efficiencies displayed by S. platensis suggests its industrial application in wastewater treatment.

Streptomyces/química , Proteínas de Bactérias/metabolismo , Águas Residuárias/química , Streptomyces/isolamento & purificação , Streptomyces/genética , Streptomyces/metabolismo , Proteínas de Bactérias/genética , Microbiologia da Água , Carbono/metabolismo , Purificação da Água , Rios/química , Floculação , Nitrogênio/metabolismo
Braz. j. microbiol ; 49(4): 832-839, Oct.-Dec. 2018. tab, graf
Artigo em Inglês | LILACS | ID: biblio-974313


ABSTRACT Clavulanic acid is a β-lactam compound with potent inhibitory activity against β-lactamases. Studies have shown that certain amino acids play essential roles in CA biosynthesis. However, quantitative evaluations of the effects of these amino acids are still needed in order to improve CA production. Here, we report a study of the nutritional requirements of Streptomyces clavuligerus for CA production. Firstly, the influence of the primary nitrogen source and the salts composition was investigated. Subsequently, soybean protein isolate was supplemented with arginine (0.0-3.20 g L-1), threonine (0.0-1.44 g L-1), ornithine (0.0-4.08 g L-1), and glutamate (0.0-8.16 g L-1), according to a two-level central composite rotatable design. A medium containing ferrous sulfate yielded CA production of 437 mg L-1, while a formulation without this salt produced only 41 mg L-1 of CA. This substantial difference suggested that Fe2+ is important for CA biosynthesis. The experimental design showed that glutamate and ornithine negatively influenced CA production while arginine and threonine had no influence. The soybean protein isolate provided sufficient C5 precursor for CA biosynthesis, so that supplementation was unnecessary. Screening of medium components, together with experimental design tools, could be a valuable way of enhancing CA titers and reducing the process costs.

Streptomyces/metabolismo , Ácido Clavulânico/biossíntese , Meios de Cultura/metabolismo , Ornitina/análise , Ornitina/metabolismo , Streptomyces/genética , Ácido Glutâmico/análise , Ácido Glutâmico/metabolismo , Meios de Cultura/química , Nitrogênio/análise , Nitrogênio/metabolismo
Braz. j. microbiol ; 49(2): 207-209, Apr.-June 2018. tab
Artigo em Inglês | LILACS | ID: biblio-889240


Abstract Streptomycetes remain as one of the important sources for bioactive products. Isolated from the mangrove forest, Streptomyces gilvigriseus MUSC 26T was previously characterised as a novel streptomycete. The high quality draft genome of MUSC 26T contained 5,213,277 bp with G + C content of 73.0%. Through genome mining, several gene clusters associated with secondary metabolites production were revealed in the genome of MUSC 26T. These findings call for further investigations into the potential exploitation of the strain for production of pharmaceutically important compounds.

Streptomyces/genética , Genoma Bacteriano , Microbiologia Ambiental , Streptomyces/isolamento & purificação , Composição de Bases , Produtos Biológicos/metabolismo , Análise de Sequência de DNA , Biologia Computacional , Áreas Alagadas , Redes e Vias Metabólicas/genética , Metabolismo Secundário
Braz. j. microbiol ; 49(1): 13-15, Jan.-Mar. 2018. tab, graf
Artigo em Inglês | LILACS | ID: biblio-889194


ABSTRACT As the largest genus in Actinobacteria family, Streptomyces species have the ability to synthesize numerous compounds of diverse structures with bioactivities. Streptomyces mangrovisoli MUSC 149T was previously isolated as a novel streptomycete from mangrove forest in east coast of Peninsular Malaysia. The high quality draft genome of MUSC 149T comprises 9,165,825 bp with G + C content of 72.5%. Through bioinformatics analysis, 21 gene clusters identified in the genome were associated with the production of bioactive secondary metabolites. The presence of these biosynthetic gene clusters in MUSC 149T suggests the potential exploitation of the strain for production of medically important compounds.

Streptomyces/isolamento & purificação , Genoma Bacteriano , Sedimentos Geológicos/microbiologia , Filogenia , Streptomyces/classificação , Streptomyces/genética , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Composição de Bases , DNA Bacteriano/genética , Dados de Sequência Molecular , Sequência de Bases , Malásia
Braz. j. microbiol ; 48(4): 612-614, Oct.-Dec. 2017. tab
Artigo em Inglês | LILACS | ID: biblio-889174


ABSTRACT Here, we show the draft genome sequence of Streptomyces sp. F1, a strain isolated from soil with great potential for secretion of hydrolytic enzymes used to deconstruct cellulosic biomass. The draft genome assembly of Streptomyces sp. strain F1 has 69 contigs with a total genome size of 8,142,296 bp and G + C 72.65%. Preliminary genome analysis identified 175 proteins as Carbohydrate-Active Enzymes, being 85 glycoside hydrolases organized in 33 distinct families. This draft genome information provides new insights on the key genes encoding hydrolytic enzymes involved in biomass deconstruction employed by soil bacteria.

Proteínas de Bactérias/genética , Genoma Bacteriano , Glicosídeo Hidrolases/genética , Microbiologia do Solo , Streptomyces/enzimologia , Streptomyces/isolamento & purificação , Proteínas de Bactérias/metabolismo , Composição de Bases , Brasil , Glicosídeo Hidrolases/metabolismo , Família Multigênica , Filogenia , Streptomyces/classificação , Streptomyces/genética
Electron. j. biotechnol ; 28: 41-46, July. 2017. tab, ilus, graf
Artigo em Inglês | LILACS | ID: biblio-1015839


Background: Streptomyces clavuligerus was the producer of clavulanic acid, claR, a pathway-specific transcriptional regulator in S. clavuligerus, positively regulates clavulanic acid biosynthesis. In this study, the promoter-less kanamycin resistance gene neo was fused with claR to obtain strain NEO from S. clavuligerus F613-1. The claR-neo fusion strain NEO was mutated using physical and chemical mutagens and then screened under high concentrations of kanamycin for high-yield producers of clavulanic acid. Results: The reporter gene neo was fused downstream of claR and used as an indicator for expression levels of claR in strain NEO. After three rounds of continuous treatment and screening, the high-yield clavulanic acid-producing strain M3-19 was obtained. In the shaking flask model, the clavulanic acid titer of M3-19 reached 4.33 g/L, which is an increase of 33% over the titer of 3.26 g/L for the starting strains S. clavuligerus F613-1 and NEO. Conclusions: Our results indicate that neo can be effectively used as a reporter for the expression of late-stage biosynthetic genes when screening for high-yield strains and that this approach has strong potential for improving Streptomyces strains of industrial value.

Streptomyces/genética , Streptomyces/metabolismo , Canamicina , Ácido Clavulânico/biossíntese , Fatores de Transcrição/genética , Transcrição Genética , Bioensaio , Proteínas Recombinantes , Cromatografia Líquida de Alta Pressão , Mutagênese , Regiões Promotoras Genéticas , Genes Reporter , Fusão Gênica , Fermentação , Reação em Cadeia da Polimerase em Tempo Real
Braz. j. microbiol ; 48(1): 32-36, Jan.-Mar. 2017. tab, graf
Artigo em Inglês | LILACS | ID: biblio-839352


Abstract Actinobacteria occur in many environments and have the capacity to produce secondary metabolites with antibiotic potential. Identification and taxonomy of actinobacteria that produce antimicrobial substances is essential for the screening of new compounds, and sequencing of the 16S region of ribosomal DNA (rDNA), which is conserved and present in all bacteria, is an important method of identification. Melanized fungi are free-living organisms, which can also be pathogens of clinical importance. This work aimed to evaluate growth inhibition of melanized fungi by actinobacteria and to identify the latter to the species level. In this study, antimicrobial activity of 13 actinobacterial isolates from the genus Streptomyces was evaluated against seven melanized fungi of the genera Exophiala, Cladosporium, and Rhinocladiella. In all tests, all actinobacterial isolates showed inhibitory activity against all isolates of melanized fungi, and only one actinobacterial isolate had less efficient inhibitory activity. The 16S rDNA region of five previously unidentified actinobacterial isolates from Ilha do Mel, Paraná, Brazil, was sequenced; four of the isolates were identified as Streptomyces globisporus subsp. globisporus, and one isolate was identified as Streptomyces aureus. This work highlights the potential of actinobacteria with antifungal activity and their role in the pursuit of novel antimicrobial substances.

Actinobacteria/fisiologia , Fungos/fisiologia , Antibiose , Filogenia , Streptomyces/classificação , Streptomyces/genética , Brasil , RNA Ribossômico 16S/genética , Actinobacteria/isolamento & purificação , Actinobacteria/classificação , Actinobacteria/genética
Arq. bras. med. vet. zootec ; 69(1): 123-129, jan.-fev. 2017. tab, graf
Artigo em Português | LILACS, VETINDEX | ID: biblio-834162


As proteases fibrinolíticas são capazes de degradar coágulos de fibrina formados dentro dos vasos sanguíneos, evitando a trombose intravascular. Em animais, a tromboflebite, que acomete frequentemente os equinos, ocasiona, em seus casos graves, a obstrução jugular e também um edema de laringe, derivando a obstrução das vias aéreas, o que possibilita um edema cerebral, ocorrendo o óbito do animal. Devido ao fato de o tratamento ser de custo elevado, faz-se necessária a investigação de outras fontesde proteases fibrinolíticas com custos menores e com menos efeitos colaterais. Diante disso, este estudo tem como objetivo produzir e caracterizar proteases fibrinolíticas obtidas de Streptomyces parvulus DPUA 1573. Para produção da enzima, foi utilizado um planejamento fatorial 24 avaliando a concentração da farinha de soja (0,5, 1,0 e 1,5%) e da glicose (0, 0,5 e 1,0g/L), temperatura (28, 32 e 37ºC) e agitação (150, 200 e 250rpm) sobre a biomassa e a atividade fibrinolítica. Pode-se verificar que a protease fibrinolítica apresentou atividade máxima (835U/mL) nas condições de concentração de 1,5% de soja, 1g/L de glicose, 28°C e 150rpm com 48 horas de fermentação. A protease fibrinolítica obtida teve temperatura e pH ótimos de 55°C e pH 9,0, respectivamente. A atividade enzimática foi inibida pelo EDTA, pelo íon Fe2+ e pelo SDS, o que indicou a enzima ser uma metaloprotease. A linhagem Streptomyces parvulus DPUA 1573 foi capaz de produzir protease fibrinolítica, possuindo características bioquímicas favoráveis à aplicação na medicina veterinária e possivelmente humana.(AU)

Fibrinolytic proteases are able to degrade fibrin clot formed in the blood vessel, avoiding intravascular thrombosis. In animals, thrombophlebitis often affects horses, and in severe cases causes obstruction of the jugular and laryngeal edema leading to airway obstruction allowing cerebral edema resulting in the death of the animal. Since treatment is costly, the investigation of other sources of fibrinolytic proteases at lower cost and with fewer side effects is needed. Thus, this study aims to produce and characterize fibrinolytic proteases from Streptomyces parvulus DPUA 1573. For enzyme production, a factorial design was performed to evaluate 24 soybean flour concentration (0.5, 1.0 and 1.5%) and glucose (0, 0.5 and 1.0g/L), temperature (28, 32 and 37°C) and agitation (150, 200 and 250rpm) on biomass and fibrinolytic activity. Fibrinolytic protease showed maximum activity (835 U/mL) under these conditions: 1.5% soybean flour, 1g/L glucose, 28°C, and 150rpm 48 hours of fermentation. The optimal temperature was 55°C and optimal pH was 9.0. Fibrinolytic protease activity was inhibited by EDTA, the ion Fe2+, and by SDS, which indicated that the enzyme is a metallo-protease. The strain Streptomyces parvulus DPUA 1573 was able to produce fibrinolytic protease with biochemical characteristics favorable for application in veterinary and human medicine.(AU)

Fermentação , Fibrinolíticos , Peptídeo Hidrolases/análise , Streptomyces , Metaloproteases
Pesqui. vet. bras ; 36(9): 805-810, set. 2016. tab, graf
Artigo em Português | LILACS, VETINDEX | ID: biblio-829311


A mastite é uma inflamação na glândula mamária que pode acarretar perdas na produção e na qualidade do leite, gerando prejuízos econômicos para a pecuária leiteira. O tratamento é baseado na utilização de antibióticos, sendo, em muitos casos, ineficazes devido à resistência bacteriana já conhecida para esta doença. O objetivo deste trabalho foi selecionar linhagens de Streptomyces spp. produtoras de biocompostos com atividade antimicrobiana frente a isolados do gênero Staphylococcus multirresistentes de búfalas com mastite. Bem como, determinar os melhores parâmetros de produção, e avaliar a produção simultânea de ácido clavulânico. A seleção de Streptomyces spp. com capacidade de produzir compostos com atividade antimicrobiana foi realizada através da técnica bloco de gelose. Dentre as 30 espécies de Streptomyces spp. testadas, o micro-organismo Streptomyces parvulus DPUA 1573 apresentou melhores resultados, sendo capaz de inibir o crescimento de 7 isolados Staphylococcus spp. multirresistentes. Posteriormente, a espécie selecionada Streptomyces parvulus DPUA 1573 foi cultivada em diferentes condições pré-determinadas pelo planejamento fatorial 24, onde as variáveis independentes foram: concentração de soja (0,5; 1,0; 1,5%), glicose (0; 0,5; 1g/L), agitação (150; 200; 250rpm) e temperatura (28; 32; 37°C) e todos os ensaios do planejamento foram monitorados até 120 horas de cultivo. Todas as variáveis independentes influenciaram positivamente no crescimento celular, enquanto que para atividade antimicrobiana apenas as variáveis temperatura e agitação apresentaram efeitos significativos positivos. O líquido metabólito produzido por Streptomyces parvulus DPUA 1573 foi capaz de inibir o crescimento de sete Staphylococcus spp. multirresistentes. As melhores condições de cultivo para a produção de moléculas bioativas por este micro-organismo foi a 37?C, com 250rpm de agitação por período de 72 horas. Nos ensaios que apresentaram atividade antimicrobiana, foi avaliada a produção de ácido clavulânico ao longo do cultivo. A maior concentração de ácido clavulânico foi de 269,84g/L obtidas nas condições de 1,5% de farinha de soja em ausência de glicose no tempo de 96 horas. A linhagem Streptomyces parvulus DPUA 1573 foi eficiente contra Staphylococcus spp. multirresistentes isolados de mastite em búfalas, ainda apresentando concomitantemente produção de ácido clavulânico com o potencial uso farmacêutico.(AU)

Mastitis is an inflammation in one or more mammary glands which can lead to reduction in production and quality of milk causing economic losses to dairy farming. The use of antibiotics is the key for the treatment of this disease, but in many cases ineffective due to bacterial resistance already known for this condition. The aim of this study was to select strains of Streptomyces spp. producing biomolecules with antimicrobial activity against multidrug-resistant Staphylococcus isolated from buffaloes with mastitis, as well as to determine the best production parameters to the evaluation of simultaneous production of clavulanic acid. Thirty species of Streptomyces spp. were used to selecting the greatest producer spectrum of antimicrobial activity (agar block technique), with selection of Streptomyces parvulus DPUA 1573, and 7 multidrug-resistant Staphylococcus spp. sensitive to its biocompounds. The selected strain of Streptomyces parvulus DPUA 1573 was cultured in different conditions predetermined by the factorial design 24, where the independent variables were: soybean concentration (0.5, 1.0, 1.5%), glucose (0, 0.5, 1g/L), agitation (150, 200, 250rpm) and temperature (28, 32, 37°C); all the tests were monitored up to 120 hours of cultivation. All independent variables influenced positively the cell growth, while for antimicrobial activity only the variables temperature and agitation showed positive effects. The antimicrobial bio compounds showed activity against seven multidrug-resistant Staphylococcus spp under the conditions: temperature 37°C, agitation 250rpm, with 72 hours of production process. In the tests which showed antimicrobial activity, was also assessed the production of clavulanic acid along with the cultivation. The highest concentration of clavulanic acid was 269.84g/L obtained under the conditions of 1.5% of soybean flour and absence of glucose in 96 hours. The strain Streptomyces parvulus DPUA 1573 was effective against multidrug-resistant strains of Staphylococcus spp. of mastitis from buffaloes, still showing concomitantly production of clavulanic acid for pharmaceutical use.(AU)

Anti-Infecciosos/análise , Resistência Microbiana a Medicamentos , Mastite Bovina , Staphylococcus , Streptomyces/imunologia , Búfalos , Ácido Clavulânico/análise
Biosci. j. (Online) ; 32(5): 1314-1323, sept./oct 2016. ilus, tab, graf
Artigo em Inglês | LILACS | ID: biblio-965726


The most practical approach to reduce morbidity and mortality of coronary heart disease (CHD) is to delay the process of thrombus by usage of clot-dissolving agents. The necessities of such safer compounds are to be critically examined for thrombolytic activity especially, from marine sources. Thrombolytic agents have been investigated as a possible treatment for thrombus. The aim of this study was to investigate the in vitro thrombolytic potential of Streptomyces sp.VITJS4 (NCIM No. 5574); (ACC No: JQ234978.1) active compounds. The fibrin degradation revealed a clear transparent zone of clearance with 500µg/mL concentration showing 24mm hydrolysis. The thrombolytic effect of Streptomyces sp.VITJS4 compounds was also demonstrated in vitro clot lysis assay where The percent of thrombolysis by the crude extract showed 90±1.7% at the concentration of 1000µg/mL, whereas percent of thrombolysis by streptokinase was found 100± 00%%. The bioactive compounds were further studied for spectrophotometric analysis. The UV-VIS profile showed different peaks ranging from 400-700 nm with different absorption respectively. The data confirmed the presence of both analogues with absorption maxima at 210 and 310 nm. A sensitive method using LC-MS technique was optimized for the separation and identification of bioactive metabolites which was indicated by the fingerprints. The results of the LC-MS analysis provided different peaks determining the presence of compounds with different therapeutic activities. The current study refers the bioactive compound as impressive thrombolytic agent for further laboratory study. Further studies should be conducted to ensure the efficacy and safety of different concentration of bioactive compounds for drug development. Hence the results reported perhaps useful for the discovery of novel thrombolytic drugs from marine origin.

A abordagem mais prática para reduzir a morbidade e a mortalidade da doença arterial coronariana (CHD, do inglês coronary heart disease) consiste em retardar o processo de trombo através da utilização de agentes de dissolução de coágulos. As necessidades de tais compostos mais seguros devem ser criticamente examinadas para a atividade trombolítica, especialmente de fontes marinhas. Agentes trombolíticos tem sido estudados como um possível tratamento para o trombo. O objetivo deste estudo foi investigar o potencial trombolítico in vitro dos compostos ativos do Streptomyces sp.VITJS4 (NCIM No. 5574); (ACC No: JQ234978.1). A degradação da fibrina revelou um clara zona livre transparente com concentração de 500µg/mL mostrando uma hidrólise de 24mm. O efeito trombolítico dos compostos de Streptomyces sp.VITJS4 também foi demonstrado no ensaio in vitro de lise dos coágulos em que a percentagem de trombólise pelo extrato bruto mostrou 90±1.7% a uma concentração de 1000µg/mL, enquanto que a percentagem de trombólise pela estreptoquinase foi de 100± 00%. Os compostos bioativos foram estudados posteriormente através da análise espectrofotométrica. O perfil ultra violeta visível (UV-VIS profile, em inglês) mostrou diferentes picos variando entre 400-700 nm com diferentes absorções respectivamente. Os dados confirmaram a presença de ambos os análogos com absorção máxima em 210 e 300 nm. Um método sensível usando a técnica LC-MS (Liquid chromatography­mass spectrometry) foi otimizado para a separação e identificação metabólitos bioativos que foram indicados pelas impressões digitais (?). Os resultados da análise LC-MS forneceram diferentes picos determinando a presença de compostos com diferentes atividades terapêuticas. O estudo atual refere-se ao composto bioativo como um agente trombolítico impressionante para futuros estudos em laboratório. Estudos futuros devem ser conduzidos para assegurar a eficácia e segurança de diferentes concentrações dos compostos bioativos para o desenvolvimento de drogas. Assim, os resultados reportados talvez sejam úteis para a descoberta de novas drogas trombolíticas de origem marinha.

Streptomyces , Trombose , Técnicas In Vitro , Actinobacteria , Fibrinolíticos
Braz. j. microbiol ; 47(3): 603-609, July-Sept. 2016. graf
Artigo em Inglês | LILACS | ID: lil-788982


ABSTRACT Streptomyces lunalinharesii strain 235 produces an antimicrobial substance that is active against sulfate reducing bacteria, the major bacterial group responsible for biofilm formation and biocorrosion in petroleum reservoirs. The use of this antimicrobial substance for sulfate reducing bacteria control is therefore a promising alternative to chemical biocides. In this study the antimicrobial substance did not interfere with the biofilm stability, but the sulfate reducing bacteria biofilm formation was six-fold smaller in carbon steel coupons treated with the antimicrobial substance when compared to the untreated control. A reduction in the most probable number counts of planktonic cells of sulfate reducing bacteria was observed after treatments with the sub-minimal inhibitory concentration, minimal inhibitory concentration, and supra-minimal inhibitory concentration of the antimicrobial substance. Additionally, when the treated coupons were analyzed by scanning electron microscopy, the biofilm formation was found to be substantially reduced when the supra-minimal inhibitory concentration of the antimicrobial substance was used. The coupons used for the biofilm formation had a small weight loss after antimicrobial substance treatment, but corrosion damage was not observed by scanning electron microscopy. The absence of the dsrA gene fragment in the scraped cell suspension after treatment with the supra-minimal inhibitory concentration of the antimicrobial substance suggests that Desulfovibrio alaskensis was not able to adhere to the coupons. This is the first report on an antimicrobial substance produced by Streptomyces active against sulfate reducing bacteria biofilm formation. The application of antimicrobial substance as a potential biocide for sulfate reducing bacteria growth control could be of great interest to the petroleum industry.

Oxirredução , Streptomyces/fisiologia , Sulfatos/metabolismo , Biofilmes , Antibiose , Streptomyces/efeitos dos fármacos , Streptomyces/ultraestrutura , Testes de Sensibilidade Microbiana , Biofilmes/crescimento & desenvolvimento , Biofilmes/efeitos dos fármacos , Antibacterianos/farmacologia
Braz. j. pharm. sci ; 52(2): 265-272, Apr.-June 2016. tab, graf
Artigo em Inglês | LILACS | ID: lil-794996


ABSTRACT This study evaluated the influence of glucose and stirring in the fermentation process in order to produce anti-Candida metabolites produced by Streptomyces sp. MPO4 isolated from Amazon soil. The anti-Candida metabolites production was registered after 24 h of fermentation in stirred ISP2 medium, having antifungal inhibition halos between 12.3 mm and 25.3 mm, yielding higher production of anti-Candida agents after 96 h. Stirring was a determining factor for the production of anti-Candida secondary metabolites, since the absence of glucose reflected in the late production of the antifungal starting from Streptomyces sp.

RESUMO Este estudo avaliou a influência da glicose e agitação no processo de fermentação para a produção de metabólitos anti-Candida produzidos por Streptomyces sp. MPO4 isolado do solo da Amazônia. A produção dos metabólitos anti-Candida foi registrada a partir de 24 h de fermentação sob agitação em meio ISP2, apresentando halos de inibição entre 12,3 mm e 25,3 mm, obtendo-se maior produção do antifúngico em 96 h. A agitação foi um fator determinante para a produção de metabólitos secundários anti-Candida e a ausência de glicose refletiu na produção tardia do antifúngico a partir do Streptomyces sp.

Candida albicans/química , Fermentação/efeitos dos fármacos , Glucose/análise , Streptomyces/química , Antifúngicos/farmacologia
Braz. j. microbiol ; 46(4): 957-968, Oct.-Dec. 2015. tab, graf
Artigo em Inglês | LILACS | ID: lil-769664


Abstract L-glutaminase was produced by Streptomyces canarius FR (KC460654) with an apparent molecular mass of 44 kDa. It has 17.9 purification fold with a final specific activity 132.2 U/mg proteins and 28% yield recovery. The purified L-glutaminase showed a maximal activity against L-glutamine when incubated at pH 8.0 at 40 °C for 30 min. It maintained its stability at wide range of pH from 5.0 11.0 and thermal stable up to 60 °C with Tm value 57.5 °C. It has high affinity and catalytic activity for L-glutamine (Km 0.129 mM, Vmax 2.02 U/mg/min), followed by L-asparagine and L-aspartic acid. In vivo, L-glutaminase showed no observed changes in liver; kidney functions; hematological parameters and slight effect on RBCs and level of platelets after 10 days of rabbit's injection. The anticancer activity of L-glutaminase was also tested against five types of human cancer cell lines using MTT assay in vitro. L-glutaminase has a significant efficiency against Hep-G2 cell (IC50, 6.8 μg/mL) and HeLa cells (IC50, 8.3 μg/mL), while the growth of MCF-7 cells was not affected. L-glutaminase has a moderate cytotoxic effect against HCT-116 cell (IC50, 64.7 μg/mL) and RAW 264.7 cell (IC50, 59.3 μg/mL).

Animais/química , Animais/efeitos dos fármacos , Animais/enzimologia , Animais/metabolismo , Animais/farmacologia , Antineoplásicos/química , Antineoplásicos/efeitos dos fármacos , Antineoplásicos/enzimologia , Antineoplásicos/metabolismo , Antineoplásicos/farmacologia , Biocatálise/química , Biocatálise/efeitos dos fármacos , Biocatálise/enzimologia , Biocatálise/metabolismo , Biocatálise/farmacologia , Proliferação de Células/química , Proliferação de Células/efeitos dos fármacos , Proliferação de Células/enzimologia , Proliferação de Células/metabolismo , Proliferação de Células/farmacologia , Estabilidade Enzimática/química , Estabilidade Enzimática/efeitos dos fármacos , Estabilidade Enzimática/enzimologia , Estabilidade Enzimática/metabolismo , Estabilidade Enzimática/farmacologia , Glutaminase/química , Glutaminase/efeitos dos fármacos , Glutaminase/enzimologia , Glutaminase/metabolismo , Glutaminase/farmacologia , Glutamina/química , Glutamina/efeitos dos fármacos , Glutamina/enzimologia , Glutamina/metabolismo , Glutamina/farmacologia , Células HeLa/química , Células HeLa/efeitos dos fármacos , Células HeLa/enzimologia , Células HeLa/metabolismo , Células HeLa/farmacologia , /química , /efeitos dos fármacos , /enzimologia , /metabolismo , /farmacologia , Humanos/química , Humanos/efeitos dos fármacos , Humanos/enzimologia , Humanos/metabolismo , Humanos/farmacologia , Cinética/química , Cinética/efeitos dos fármacos , Cinética/enzimologia , Cinética/metabolismo , Cinética/farmacologia , Streptomyces/química , Streptomyces/efeitos dos fármacos , Streptomyces/enzimologia , Streptomyces/metabolismo , Streptomyces/farmacologia , Especificidade por Substrato/química , Especificidade por Substrato/efeitos dos fármacos , Especificidade por Substrato/enzimologia , Especificidade por Substrato/metabolismo , Especificidade por Substrato/farmacologia
Int. braz. j. urol ; 41(2): 367-372, Mar-Apr/2015. tab, graf
Artigo em Inglês | LILACS | ID: lil-748287


Objective The aim of active surveillance of early prostate cancer is to individualize therapy by selecting for curative treatment only patients with significant cancer. Epstein’s criteria for prediction of clinically insignificant cancer in surgical specimens are widely used. Epstein’s criterion “no single core with >50% cancer” has no correspondence in linear extent. The aim of this study is to find a possible correspondence. Materials and Methods From a total of 401 consecutive patients submitted to radical prostatectomy, 17 (4.2%) met criteria for insignificant cancer in the surgical specimen. The clinicopathologic findings in the correspondent biopsies were compared with Epstein’s criteria for insignificant cancer. Cancer in a single core was evaluated in percentage as well as linear extent in mm. Results Comparing the clinicopathologic findings with Epstein’s criteria predictive of insignificant cancer, there was 100% concordance for clinical stage T1c, no Gleason pattern 4 or 5, ≤2 cores with cancer, and no single core with >50% cancer. However, only 25% had density ≤0.15. The mean, median and range of the maximum length of cancer in a single core in mm were 1.19, 1, and 0.5-2.5, respectively. Additionally, the mean, median, and range of length of cancer in all cores in mm were 1.47, 1.5, and 0.5-3, respectively. Conclusion To pathologists that use Epstein’s criteria predictive of insignificant cancer and measure linear extent in mm, our study favors that “no single core with >50% cancer” may correspond to >2.5 mm in linear extent. .

Policetídeo Sintases/química , Policetídeo Sintases/ultraestrutura , Streptomyces/enzimologia , Biocatálise , Domínio Catalítico , Microscopia Crioeletrônica , Ácido Graxo Sintases/química , Modelos Moleculares , Macrolídeos/metabolismo , Policetídeo Sintases/metabolismo
Braz. j. microbiol ; 45(4): 1179-1186, Oct.-Dec. 2014. graf, tab
Artigo em Inglês | LILACS | ID: lil-741267


Two mesophilic streptomycetes (S. violaceoruber and S. spiroverticillatus) were selected to study their Poly R-478 decolorization ability and lignocellulose solubilizing activity. Both strains were able to degrade Poly R-478 dye and ferulic acid during growth on a minimal salts medium. The Poly R-478 decolorizing activities of both strains were induced by adding different carbon sources to the culture media. S. violaceoruber could decolorize 63% of Poly R-478 after 24 h. Both strains could solubilize straw and produce acid-precipitable polymeric lignin (APPL) with different efficiency. From the major extracellular enzymes recovery of both strains on rice and wheat straw, we can predicate that the biodegradation process was partial indicating a possible utilization in biological delignification.

Antraquinonas/metabolismo , Lignina/metabolismo , Polímeros/metabolismo , Streptomyces/metabolismo , Biotransformação , Ácidos Cumáricos/metabolismo , Meios de Cultura/química , Oryza/metabolismo , Caules de Planta/metabolismo , Streptomyces/crescimento & desenvolvimento , Triticum/metabolismo
Braz. j. microbiol ; 45(3): 919-928, July-Sept. 2014. graf, tab
Artigo em Inglês | LILACS | ID: lil-727021


The selection of new microorganisms able to produce antimicrobial compounds is hoped for to reduce their production costs and the side effects caused by synthetic drugs. Clavulanic acid is a β-lactam antibiotic produced by submerged culture, which is widely used in medicine as a powerful inhibitor of β-lactamases, enzymes produced by bacteria resistant to antibiotics such penicillin and cephalosporin. The purpose of this work was to select the best clavulanic acid producer among strains of Streptomyces belonging to the Microorganism Collection of the Department of Antibiotics of the Federal University of Pernambuco (DAUFPE). Initially, the strains were studied for their capacity to inhibit the action of β-lactamases produced by Klebsiella aerogenes ATCC 15380. From these results, five strains were selected to investigate the batch kinetics of growth and clavulanic acid production in submerged culture carried out in flasks. The results were compared with the ones obtained by Streptomyces clavuligerus ATCC 27064 selected as a control strain. The best clavulanic acid producer was Streptomyces DAUFPE 3060, molecularly identified as Streptomyces variabilis, which increased the clavulanic acid production by 28% compared to the control strain. This work contributes to the enlargement of knowledge on new Streptomyces wild strains able to produce clavulanic acid by submerged culture.

Ácido Clavulânico/metabolismo , Inibidores Enzimáticos/metabolismo , Streptomyces/isolamento & purificação , Streptomyces/metabolismo , Enterobacter aerogenes/enzimologia , Programas de Rastreamento , Streptomyces/crescimento & desenvolvimento , beta-Lactamases/metabolismo
Rev. bras. hematol. hemoter ; 36(4): 297-299, Jul-Aug/2014. graf
Artigo em Inglês | LILACS | ID: lil-718392


A 29-year-old woman with stage IVB Hodgkin's lymphoma was treated with doxorubicin, bleomycin, vinblastine, and dacarbazine. Two weeks after the first cycle was completed, she developed pruritic, linear erythematous lesions in a flagellate-like pattern on the trunk, neck and arms. After oral prednisone therapy and cessation of bleomycin, the lesions started to recede...

Humanos , Feminino , Adulto , Bleomicina , Dermatite , Eritema , Doença de Hodgkin , Streptomyces