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1.
J Antibiot (Tokyo) ; 75(11): 610-618, 2022 11.
Artigo em Inglês | MEDLINE | ID: mdl-36076014

RESUMO

Four novel cyclic enaminones, designated RD4123A-D (1-4), and a new 4-quinazolinone metabolite, RD4123E (5), were isolated from the culture extract of an unidentified actinomycete strain RD004123, which belongs to the family Micromonosporaceae. Structures of 1-5 were determined by spectroscopic analyses using NMR, MS, and electronic circular dichroism (ECD), combined with quantum chemical calculations of ECD and NMR chemical shifts and biosynthetic consideration. Compounds 1-5 showed weak to modest cytotoxicity against murine leukemia P388 cells, while being inactive against bacteria and fungi.


Assuntos
Actinobacteria , Micromonosporaceae , Actinobacteria/química , Animais , Dicroísmo Circular , Camundongos , Extratos Vegetais , Quinazolinonas
2.
Molecules ; 27(5)2022 Mar 07.
Artigo em Inglês | MEDLINE | ID: mdl-35268843

RESUMO

The COVID-19 pandemic has led to the search for new molecules with antiviral activity against SARS-CoV-2. The entry of the virus into the cell is one of the main targets for inhibiting SARS-CoV-2 infection. Natural products are an important source of new therapeutic alternatives against diseases. Pseudotyped viruses allow the study of SARS-CoV-2 viral entry inhibitors, and due to their simplicity, they allow the screening of a large number of antiviral candidates in Biosafety Level 2 facilities. We used pseudotyped HIV-1 with the D614G SARS-CoV-2 spike glycoprotein to test its ability to infect ACE2-expressing HEK 293T cells in the presence of diverse natural products, including 21 plant extracts, 7 essential oils, and 13 compounds from plants and fungi. The 50% cytotoxic concentration (CC50) was evaluated using the resazurin method. From these analyses, we determined the inhibitory activity of the extract of Stachytarpheta cayennensis, which had a half-maximal inhibitory concentration (IC50) of 91.65 µg/mL, a CC50 of 693.5 µg/mL, and a selectivity index (SI) of 7.57, indicating its potential use as an inhibitor of SARS-CoV-2 entry. Moreover, our work indicates the usefulness of the pseudotyped-virus system in the screening of SARS-CoV-2 entry inhibitors.


Assuntos
Antivirais/farmacologia , Produtos Biológicos/química , Internalização do Vírus/efeitos dos fármacos , Actinobacteria/química , Actinobacteria/metabolismo , Enzima de Conversão de Angiotensina 2/genética , Enzima de Conversão de Angiotensina 2/metabolismo , Antivirais/química , Antivirais/metabolismo , Antivirais/uso terapêutico , Produtos Biológicos/metabolismo , Produtos Biológicos/farmacologia , Produtos Biológicos/uso terapêutico , COVID-19/virologia , Células HEK293 , Ensaios de Triagem em Larga Escala/métodos , Humanos , Óleos Voláteis/química , Óleos Voláteis/farmacologia , Óleos Voláteis/uso terapêutico , Extratos Vegetais/química , Extratos Vegetais/metabolismo , Extratos Vegetais/farmacologia , SARS-CoV-2/isolamento & purificação , SARS-CoV-2/fisiologia , Glicoproteína da Espícula de Coronavírus/antagonistas & inibidores , Glicoproteína da Espícula de Coronavírus/metabolismo , Tratamento Farmacológico da COVID-19
3.
ACS Chem Biol ; 16(5): 915-928, 2021 05 21.
Artigo em Inglês | MEDLINE | ID: mdl-33913701

RESUMO

Glycopeptide antibiotics (GPAs) are last defense line drugs against multidrug-resistant Gram-positive pathogens. Natural GPAs teicoplanin and vancomycin, as well as semisynthetic oritavancin, telavancin, and dalbavancin, are currently approved for clinical use. Although these antibiotics remain efficient, emergence of novel GPA-resistant pathogens is a question of time. Therefore, it is important to investigate the natural variety of GPAs coming from so-called "rare" actinobacteria. Herein we describe a novel GPA producer-Nonomuraea coxensis DSM 45129. Its de novo sequenced and completely assembled genome harbors a biosynthetic gene cluster (BGC) similar to the dbv BGC of A40926, the natural precursor to dalbavancin. The strain produces a novel GPA, which we propose is an A40926 analogue lacking the carboxyl group on the N-acylglucosamine moiety. This structural difference correlates with the absence of dbv29-coding for an enzyme responsible for the oxidation of the N-acylglucosamine moiety. Introduction of dbv29 into N. coxensis led to A40926 production in this strain. Finally, we successfully applied dbv3 and dbv4 heterologous transcriptional regulators to trigger and improve A50926 production in N. coxensis, making them prospective tools for screening other Nonomuraea spp. for GPA production. Our work highlights genus Nonomuraea as a still untapped source of novel GPAs.


Assuntos
Actinobacteria/química , Antibacterianos/química , Proteínas de Bactérias/química , Glicopeptídeos/química , Proteínas Recombinantes/química , Actinobacteria/genética , Antibacterianos/farmacologia , Proteínas de Bactérias/genética , Proteínas de Bactérias/farmacologia , Sequência de Bases , Simulação por Computador , Avaliação Pré-Clínica de Medicamentos , Regulação Bacteriana da Expressão Gênica , Genômica/métodos , Glucosamina/química , Glicopeptídeos/farmacologia , Família Multigênica , Proteínas Recombinantes/genética , Proteínas Recombinantes/farmacologia , Espectrometria de Massas em Tandem , Teicoplanina/análogos & derivados , Teicoplanina/química , Teicoplanina/farmacologia
4.
Drug Discov Ther ; 14(6): 287-295, 2021 Jan 23.
Artigo em Inglês | MEDLINE | ID: mdl-33250497

RESUMO

In vivo-mimic silkworm infection models with Mycobacterium avium and Mycobacterium intracellulare were newly established to evaluate the therapeutic effects of anti-M. avium complex (MAC) antibiotics. Silkworms raised at 37°C died within 72 hours of an injection of M. avium or M.intracellulare (2.5 × 107 colony-forming unit (CFU)/larva·g) into the hemolymph. Clinical anti-mycobacterial (tuberculosis) antibiotics were evaluated under these conditions. Clarithromycin, kanamycin, streptomycin, amikacin, and ciprofloxacin exerted therapeutic effects in a dose-dependent manner, which was consistent with those in the mouse model. Furthermore, three effective actinomycete culture broths were selected in the screening program of our microbial broth library using the silkworm model, and four active metabolites, ohmyungsamycins A and B (1 and 2), chartreusin (3), and griseoviridin (4), were identified. Among these compounds, 1 showed the lowest 50% effective dose (ED50) value (8.5 µg/larva·g), while 3 had the best ED50/minimum inhibitory concentration (MIC) ratio (7.4). These results indicate that silkworm models are a useful tool for identifying anti-MAC antibiotics candidates with veritable therapeutic effects.


Assuntos
Actinobacteria/química , Antibacterianos/administração & dosagem , Bombyx/microbiologia , Complexo Mycobacterium avium/patogenicidade , Infecção por Mycobacterium avium-intracellulare/tratamento farmacológico , Animais , Antibacterianos/farmacologia , Benzopiranos/administração & dosagem , Benzopiranos/farmacologia , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Avaliação Pré-Clínica de Medicamentos , Glicosídeos/administração & dosagem , Glicosídeos/farmacologia , Testes de Sensibilidade Microbiana , Complexo Mycobacterium avium/efeitos dos fármacos , Complexo Mycobacterium avium/crescimento & desenvolvimento , Peptídeos/administração & dosagem , Peptídeos/farmacologia , Peptídeos Cíclicos/administração & dosagem , Peptídeos Cíclicos/farmacologia
5.
Front Immunol ; 11: 1548, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32733487

RESUMO

Background: The COVID-19 pandemic has been causing varying severities of illness. Some are asymptomatic and some develop severe disease leading to mortality across ages. This contrast triggered us explore the causes, with the background that a vaccine for effective immunization or a drug to tackle COVID-19 is not too close to reality. We have discussed strategies to combat COVID-19 through immune enhancement, using simple measures including nutritional supplements. Discussion: A literature search on mortality-related comorbid conditions was performed. For those conditions, we analyzed the pro-inflammatory cytokines, which could cause the draining of the immune reservoir. We also analyzed the immune markers necessary for the defense mechanism/immune surveillance against COVID-19, especially through simple means including immune enhancing nutritional supplement consumption, and we suggest strategies to combat COVID-19. Major comorbid conditions associated with increased mortality include cardiovascular disease (CVD), diabetes, being immunocompromised by cancer, and severe kidney disease with a senile immune system. Consumption of Aureobasidium pullulans strain (AFO-202) beta 1,3-1,6 glucan supported enhanced IL-8, sFAS macrophage activity, and NK cells' cytotoxicity, which are major defense mechanisms against viral infection. Conclusion: People with co-morbid conditions who are more prone to COVID-19-related deaths due to immune dysregulation are likely to benefit from consuming nutritional supplements that enhance the immune system. We recommend clinical studies to validate AFO-202 beta glucan in COVID-19 patients to prove its efficacy in overcoming a hyper-inflammation status, thus reducing the mortality, until a definite vaccine is made available.


Assuntos
Betacoronavirus , Doenças Cardiovasculares/epidemiologia , Infecções por Coronavirus/epidemiologia , Infecções por Coronavirus/imunologia , Diabetes Mellitus/epidemiologia , Suplementos Nutricionais , Neoplasias/epidemiologia , Pneumonia Viral/epidemiologia , Pneumonia Viral/imunologia , Insuficiência Renal Crônica/epidemiologia , Actinobacteria/química , Biomarcadores/sangue , COVID-19 , Doenças Cardiovasculares/imunologia , Comorbidade , Infecções por Coronavirus/dietoterapia , Infecções por Coronavirus/mortalidade , Citocinas/sangue , Diabetes Mellitus/imunologia , Humanos , Hospedeiro Imunocomprometido , Neoplasias/imunologia , Pandemias , Pneumonia Viral/dietoterapia , Pneumonia Viral/mortalidade , Insuficiência Renal Crônica/imunologia , SARS-CoV-2 , beta-Glucanas/farmacologia , beta-Glucanas/uso terapêutico
6.
J Biosci Bioeng ; 130(1): 106-113, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32253091

RESUMO

Monoclonal antibodies (mAbs) are active pharmaceutical ingredients in antibody drugs, produced mainly using recombinant Chinese hamster ovary (CHO) cells. The regulation of recombinant CHO cell proliferation can improve the productivity of heterologous proteins. Chemical compound approaches for cell cycle regulation have the advantages of simplicity and ease of use in industrial processes. However, CHO cells have genetic and phenotypic diversity, and the effects of such compounds might depend on cell line and culture conditions. Increasing the variety of cell cycle inhibitors is a promising strategy to overcome the dependency. Marine microorganisms are a vast and largely undeveloped source of secondary metabolites with physiological activity. In this study, we focused on secondary metabolites of marine microorganisms and evaluated their effectiveness as cell cycle inhibitory compounds. Of 720 extracts from microorganisms (400 actinomycetes and 320 filamentous fungi) collected from the Okinawan Sea, we identified nine extracts that decreased the specific growth rate and increased the specific production rate without reducing cell viability. After fractionating the extracts, the components of active fractions were estimated using time-of-flight mass spectrometry analysis. Then, four compounds, including staurosporine and undecylprodigiosin were deduced to be active compounds. These compounds have been reported to exert a cell cycle inhibitory effect on mammalian cells. These compounds might serve as additives to improve mAb production in CHO cells. This study indicates that secondary metabolites of marine microorganisms are a useful source for new cell cycle inhibitory compounds that can increase mAb production in CHO cells.


Assuntos
Actinobacteria/química , Ciclo Celular/efeitos dos fármacos , Fungos/química , Inibidores do Crescimento/farmacologia , Água do Mar/microbiologia , Actinobacteria/genética , Actinobacteria/isolamento & purificação , Actinobacteria/metabolismo , Animais , Células CHO , Divisão Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Cricetinae , Cricetulus , Avaliação Pré-Clínica de Medicamentos , Fungos/genética , Fungos/isolamento & purificação , Fungos/metabolismo , Inibidores do Crescimento/metabolismo , Prodigiosina/análogos & derivados , Prodigiosina/metabolismo , Prodigiosina/farmacologia , Estaurosporina/metabolismo , Estaurosporina/farmacologia
7.
Nutr Res Rev ; 33(2): 190-217, 2020 12.
Artigo em Inglês | MEDLINE | ID: mdl-32051057

RESUMO

Ergothioneine (ERG) is an unusual thio-histidine betaine amino acid that has potent antioxidant activities. It is synthesised by a variety of microbes, especially fungi (including in mushroom fruiting bodies) and actinobacteria, but is not synthesised by plants and animals who acquire it via the soil and their diet, respectively. Animals have evolved a highly selective transporter for it, known as solute carrier family 22, member 4 (SLC22A4) in humans, signifying its importance, and ERG may even have the status of a vitamin. ERG accumulates differentially in various tissues, according to their expression of SLC22A4, favouring those such as erythrocytes that may be subject to oxidative stress. Mushroom or ERG consumption seems to provide significant prevention against oxidative stress in a large variety of systems. ERG seems to have strong cytoprotective status, and its concentration is lowered in a number of chronic inflammatory diseases. It has been passed as safe by regulatory agencies, and may have value as a nutraceutical and antioxidant more generally.


Assuntos
Antioxidantes/farmacologia , Produtos Biológicos/farmacologia , Suplementos Nutricionais , Ergotioneína/farmacologia , Estresse Oxidativo/efeitos dos fármacos , Actinobacteria/química , Animais , Fungos/química , Humanos , Proteínas de Transporte de Cátions Orgânicos/metabolismo , Simportadores/metabolismo
8.
J Gen Appl Microbiol ; 66(1): 24-31, 2020 Apr 13.
Artigo em Inglês | MEDLINE | ID: mdl-31378748

RESUMO

Endophytic microbes associated with medicinal plants are considered to be potential producers of various bioactive secondary metabolites. The present study investigated the distribution, antimicrobial activity and genetic features of endophytic actinomycetes isolated from the medicinal plant Cinnamomum cassia Presl collected in Hoa Binh province of northern Vietnam. Based on phenotypic characteristics, 111 actinomycetes were isolated from roots, stems and leaves of the host plants by using nine selective media. The isolated actinomycetes were mainly recovered from stems (n = 67; 60.4%), followed by roots (n = 29; 26.1%) and leaves (n = 15; 13.5%). The isolates were accordingly assigned into 5 color categories of aerial mycelium, of which gray is the most dominant (n = 42; 37.8%), followed by white (n = 33; 29.7%), yellow (n = 25; 22,5%), red (n = 8; 7.2%) and green (n = 3; 2.7%). Of the total endophytic actinomycetes tested, 38 strains (occupying 34.2%) showed antimicrobial activity against at least one of nine tested microbes and, among them, 26 actinomycetes (68.4%) revealed anthracycline-like antibiotics production. Analysis of 16S rRNA gene sequences deposited on GenBank (NCBI) of the antibiotic-producing actinomycetes identified 3 distinct genera, including Streptomyces, Microbacterium, and Nocardia, among which Streptomyces genus was the most dominant and represented 25 different species. Further genetic investigation of the antibiotic-producing actinomycetes found that 28 (73.7%) and 11 (28.9%) strains possessed genes encoding polyketide synthase (pks) and nonribosomal peptide synthetase (nrps), respectively. The findings in the present study highlighted endophytic actinomycetes from C. cassia Presl which possessed broad-spectrum bioactivities with the potential for applications in the agricultural and pharmaceutical sectors.


Assuntos
Actinobacteria/química , Actinobacteria/classificação , Antibiose , Cinnamomum aromaticum/microbiologia , Actinobacteria/isolamento & purificação , Endófitos/química , Endófitos/classificação , Endófitos/isolamento & purificação , Peptídeo Sintases/genética , Filogenia , Plantas Medicinais/microbiologia , Policetídeo Sintases/genética , RNA Ribossômico 16S/genética , Metabolismo Secundário , Análise de Sequência de DNA , Vietnã
9.
Bioorg Chem ; 95: 103507, 2020 01.
Artigo em Inglês | MEDLINE | ID: mdl-31884136

RESUMO

Three pyrrol-2-aldehyde derivatives, including one new compound, jiangrine G (JG), two known compounds, jiangrine A (JA), and pyrrolezanthine (PZ), were isolated from the fermentation broth of Jiangella alba associated with a traditional Chinese medicinal plant Maytenus austroyunnanensis. The structure of jiangrine G was elucidated by a detailed spectroscopic data analysis including data from CD spectra. The anti-inflammatory activities assay demonstrated that JG and JA suppressed the production of pro-inflammatory cytokines including NO, IL-1ß, and IL-6 as well as inhibited the expression of iNOS in LPS-induced RAW 264.7 cells in a dose-dependent manner. While high concentration of PZ dramatically suppressed the protein expression of TNF-α, but stimulated the release of IL-1ß and IL-6. Western blot results revealed that JG, JA, and PZ modulated the expression of pro-inflammatory cytokines via MAPK p38 and NF-κB signaling pathways. For the unique structure of PZ, difference from JG and JA, the signaling pathway involved in mediating its effects on regulating the synthesis of IL-1ß and IL-6 are probably more complicated than that of JG and JA.


Assuntos
Actinobacteria/química , Anti-Inflamatórios não Esteroides/farmacologia , Inibidores Enzimáticos/farmacologia , NF-kappa B/antagonistas & inibidores , Pirróis/farmacologia , Proteínas Quinases p38 Ativadas por Mitógeno/antagonistas & inibidores , Animais , Anti-Inflamatórios não Esteroides/química , Anti-Inflamatórios não Esteroides/isolamento & purificação , Sobrevivência Celular/efeitos dos fármacos , Citocinas/antagonistas & inibidores , Citocinas/metabolismo , Relação Dose-Resposta a Droga , Inibidores Enzimáticos/química , Inibidores Enzimáticos/isolamento & purificação , Lipopolissacarídeos/antagonistas & inibidores , Lipopolissacarídeos/farmacologia , Camundongos , Estrutura Molecular , NF-kappa B/metabolismo , Óxido Nítrico Sintase Tipo II/antagonistas & inibidores , Óxido Nítrico Sintase Tipo II/metabolismo , Pirróis/química , Pirróis/isolamento & purificação , Células RAW 264.7 , Relação Estrutura-Atividade , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo
10.
Pest Manag Sci ; 75(6): 1585-1593, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30461185

RESUMO

BACKGROUND: Bursaphelenchus xylophilus is a migratory endoparasitic nematode known to cause severe environmental damage and economic losses in pine forest ecosystems. This present study investigated the nematicidal metabolites of actinomycetes in vitro and evaluated the disease control efficacy of the active compound and metabolites under greenhouse and field conditions. RESULTS: Five thousand types of actinobacteria from Korean forest soil samples were screened to identify novel nematicidal agents against the pine wood nematode. Streptomyces sp. AN091965 showed the strongest nematicidal activity. One active compound, spectinabilin, was obtained by nematicidal asssy-directed fractionation, and it showed significant nematicidal activity against B. xylophilus, with an LC50 value of 0.84 µg mL-1 . Spectinabilin effectively suppressed the development of pine wilt disease in 5-year-old Pinus densiflora trees, even at 0.9 mg per tree under greenhouse conditions. Moreover, the acetone extract of the active strain's mycelia efficiently suppressed the development of pine wilt disease under field conditions. CONCLUSION: To the best of our knowledge, this the first report to describe the nematicidal activity of spectinabilin against B. xylophilus. The cell extracts described herein merit further field studies as potential nematicides against the pine wood nematode. © 2018 Society of Chemical Industry.


Assuntos
Actinobacteria/química , Antinematódeos/isolamento & purificação , Antinematódeos/farmacologia , Nematoides/efeitos dos fármacos , Animais , Avaliação Pré-Clínica de Medicamentos
11.
Biomed Res Int ; 2018: 1470305, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30519568

RESUMO

Endophytic actinomycetes are a promising source of novel metabolites with diverse biological activities. Tea plants (Camellia sinensis) produce arsenals of phytochemicals, which are linked to a number of medicinal and nutritional properties. However, a systematic investigation into the abundance and diversity of cultivated actinomycetes residing in tea plants has not been performed. In this study, a total of 46 actinobacteria were recovered from leaf, stem, and root samples of 15 tea cultivars collected in Fujian province, China. Their abundance and diversity were shown to be influenced by both the genotypes and tissue types of tea plants. Based on 16S RNA sequence analysis, these isolates were taxonomically grouped into 11 families and 13 genera, including Streptomyces, Actinomadura, Kribbella, Nocardia, Kytococcus, Leifsonia, Microbacterium, Micromonospora, Mobilicoccus, Mycobacterium, Nocardiopsis, Piscicoccus, and Pseudonocardia. The genus Streptomyces was most prevalent whereas rare genera, Mobilicoccus and Piscicoccus, were reported for the first time to occur as plant endophytes. PCR screening of polyketide synthase genes (PKS-I and PKS-II) and nonribosomal peptide synthetase genes (NRPS), along with antimicrobial assays against a set of bacterial and fungal pathogens, showed that endophytic actinomycetes associated with tea plants have a high potential for producing antimicrobial metabolites. Furthermore, indole acetic acid (IAA) production and 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase activities were recorded in 93.5% and 21.7% of all isolates, respectively. Overall, these results indicate that endophytic actinomycetes from tea plants represent a valuable source of bioactive metabolites with antibacterial, antifungal, and plant-growth-promoting properties.


Assuntos
Actinobacteria/isolamento & purificação , Antibacterianos/isolamento & purificação , Endófitos/isolamento & purificação , Chá/microbiologia , Actinobacteria/química , Actinobacteria/classificação , Actinobacteria/genética , Aminoácidos Cíclicos/química , Antibacterianos/química , Antibacterianos/farmacologia , China , Endófitos/química , Ácidos Indolacéticos/química , Filogenia , Desenvolvimento Vegetal/efeitos dos fármacos , RNA Ribossômico 16S/genética
12.
Braz. J. Vet. Res. Anim. Sci. (Online) ; 55(3): e141243, Outubro 25, 2018. graf, tab
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-969229

RESUMO

Actinobacteria have been researched as a source that produces crude extracts, which contain bioactive compounds able to act as antimicrobial agents. The present investigation evaluated the dose-response effect of two crude extracts, obtained at Caatinga rhizosphere (Caat) and Rhizophora mangle (AMC), on in vitro ruminal fermentation by:cumulative gas production, digestibility of dry (IVDMD) and organic matter (IVOMD), and short-chain fatty acids concentration (SCFA). Three multiparous Holstein dairy cows with ruminal fistula were used as the inoculum donors and fed a basal diet consisting of corn silage, soybean meal, urea, ground corn and mineral supplement. Ruminal fluid samples were incubated in glass bottles with 1 g of the dried and milled diet, a buffer solution, and the crude extracts evaluated in four doses (0.3, 0.6, 0.9 and 1.20 mg/10 mL inoculum) in a randomized block design, and the donators were considered as blocks with random effects. Additionally, negative controls were used. The results were expressed as average values based on triplicate analyses. Decreased cumulative gas production was observed according to linear dose response at 24, 48 and 72 h of incubation with the addition of Caat extract. The IVOMD showed a linear decrease at 72 h of incubation with dose Caat inclusion. Furthermore, the inclusion of Caat extract linearly reduced butyric and isovaleric acid concentrations, as well as acetate:propionate ratio. Finally, the Caat inclusion increased the propionic acid concentration in comparison to AMC extract. However, the inclusion of AMC extract did not affect any of the analyzed variables at the used doses. The Caat extract could be used as a modulator of in vitro ruminal fermentation, since it reduced acetate:propionate ratio and cumulative gas production.(AU)


As actinobactérias têm sido pesquisadas como fonte produtoras de extratos brutos que contêm compostos bioativos capazes de atuar como agentes antimicrobianos. O presente trabalho investigou o efeito dose-resposta de dois extratos brutos, AMC e Caat, na fermentação ruminal in vitro por: produção cumulativa de gás, digestibilidade in vitro da matéria seca (IVDMD) e matéria orgânica (IVOMD) e concentração de ácidos graxos de cadeia curta (SCFA). Três vacas leiteiras da raça Holandesa, multíparas e portadoras de fístula ruminal foram utilizadas como doadoras de inóculo ruminal e foram alimentadas com uma dieta basal composta por silagem de milho, farelo de soja, ureia, milho moído e suplemento mineral. As amostras de inóculo ruminal foram incubadas em garrafas de vidro com 1 g da dieta seca e moída, solução tampão e os extratos brutos avaliados em quatro doses (0,3, 0,6, 0,9 e 1,20 mg/10 mL de inóculo) em delineamento em blocos casualizados, sendo as doadoras consideradas os blocos como efeito aleatório. Além disso, foram utilizados controles negativos para a correção da produção de gás. Os resultados foram expressos como valores médios com base em análises triplicadas. A diminuição da produção cumulativa de gás foi observada de acordo com a dose em resposta linear às 24, 48 e 72 h de incubação com a adição de extrato de Caat. A IVOMD mostrou uma diminuição linear com 72 h de incubação com inclusão de Caat. Além disso, a inclusão do Caat reduziu linearmente as concentrações de ácido butírico e isovalérico, bem como a proporção de acetato/propionato. Diferentemente, a inclusão do extrato de AMC não afetou nenhuma das variáveis analisadas nas doses utilizadas. O extrato de Caat pode ser usado como um modulador da fermentação ruminal in vitro, uma vez que reduziu a proporção de acetato/propionato e a produção de gás acumulada. (AU)


Assuntos
Actinobacteria/química , Fermentação , Ionóforos/síntese química
13.
Dokl Biochem Biophys ; 483(1): 379-381, 2018 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-30607743

RESUMO

This is the first study to detect the effect of calcium ions on the activity of transmembrane adenylyl cyclase (tmAC), the key enzyme of the adenylyl cyclase signaling system, under normal conditions and after a short-term exposure to exopolysaccharides (EPS) of the bacterial ring rot pathogen Clavibacter michiganensis ssp. sepedonicus (Cms). After the treatment of the roots of plants with the Cms EPS, the response to Ca2+ changed: the activity of the tmAC of plants of the resistant cultivar significantly increased, whereas in the cells of the susceptible cultivar it remained unchanged.


Assuntos
Actinobacteria/química , Adenilil Ciclases/metabolismo , Sinalização do Cálcio/efeitos dos fármacos , Cálcio/metabolismo , Raízes de Plantas , Polissacarídeos Bacterianos/farmacologia , Solanum tuberosum , Raízes de Plantas/citologia , Raízes de Plantas/enzimologia , Polissacarídeos Bacterianos/química , Solanum tuberosum/citologia , Solanum tuberosum/enzimologia
14.
Microb Pathog ; 110: 93-99, 2017 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-28647504

RESUMO

The aim of the present study was to isolate and evaluate the antimicrobial potential of soil actinomycetes of Kashmir Himalayas. The secondary metabolites of actinomycetes are the prominent source of antibiotics. A total of 121 morphologically different actinomycete strains were isolated and screened for antimicrobial activity against various human pathogens. The ethyl acetate extract of fermented broth an actinomycete strain, identified as Streptomyces pratensis exhibited significant antimicrobial activity against Staphylococcus aureus ATCC 29213 with MIC 0.25 µg/ml and Mycobacterium tuberculosis Strain H37Rv with MIC 0.062 µg/ml. The strain S. pratensis IIIM06 was grown on large scale and their broth was extracted with ethyl acetate. The extract was subjected to various chromatography techniques which led to the isolation of four compounds whose structures were established as actinomycin C1, actinomycin C2, actinomycin C3 and actiphenol on the basis of spectral data analysis. Actinomycin C1, C2 and C3 exhibited potent antimicrobial activity against S. aureus as well as M. tuberculosis. The isolated indigenous actinomycetes exhibited good antibacterial activity and the study reveals that IIIM06 is a promising strain and could be of great potential for industrial applications.


Assuntos
Actinobacteria/química , Actinobacteria/isolamento & purificação , Anti-Infecciosos/isolamento & purificação , Anti-Infecciosos/farmacologia , Microbiologia do Solo , Actinobacteria/classificação , Actinobacteria/genética , Anti-Infecciosos/química , DNA Bacteriano/genética , Dactinomicina/análogos & derivados , Dactinomicina/química , Dactinomicina/isolamento & purificação , Dactinomicina/farmacologia , Avaliação Pré-Clínica de Medicamentos , Fermentação , Índia , Testes de Sensibilidade Microbiana , Mycobacterium tuberculosis/efeitos dos fármacos , Filogenia , RNA Ribossômico 16S/genética , Solo , Staphylococcus aureus/efeitos dos fármacos , Streptomyces/química , Streptomyces/classificação , Streptomyces/genética , Streptomyces/isolamento & purificação
15.
Molecules ; 22(5)2017 May 19.
Artigo em Inglês | MEDLINE | ID: mdl-28534845

RESUMO

Minor ginsenosides, such as compound K, Rg3(S), which can be produced by deglycosylation of ginsenosides Rb1, showed strong anti-cancer effects. However, the anticancer effects of gypenoside LXXV, which is one of the deglycosylated shapes of ginsenoside Rb1, is still unknown due to the rarity of its content in plants. Here, we cloned and characterized a novel ginsenoside-transforming ß-glucosidase (BglG167b) derived from Microbacterium sp. Gsoil 167 which can efficiently hydrolyze gypenoside XVII into gypenoside LXXV, and applied it to the production of gypenoside LXXV at the gram-scale with high specificity. In addition, the anti-cancer activity of gypenoside LXXV was investigated against three cancer cell lines (HeLa, B16, and MDA-MB231) in vitro. Gypenoside LXXV significantly reduced cell viability, displaying an enhanced anti-cancer effect compared to gypenoside XVII and Rb1. Taken together, this enzymatic method would be useful in the preparation of gypenoside LXXV for the functional food and pharmaceutical industries.


Assuntos
Actinobacteria/química , Antineoplásicos Fitogênicos/metabolismo , Proteínas de Bactérias/metabolismo , Ginsenosídeos/metabolismo , beta-Glucosidase/metabolismo , Actinobacteria/enzimologia , Animais , Antineoplásicos Fitogênicos/isolamento & purificação , Antineoplásicos Fitogênicos/farmacologia , Proteínas de Bactérias/genética , Biotransformação , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Clonagem Molecular , Escherichia coli/genética , Escherichia coli/metabolismo , Expressão Gênica , Ginsenosídeos/isolamento & purificação , Ginsenosídeos/farmacologia , Gynostemma , Células HeLa , Humanos , Melanoma Experimental/tratamento farmacológico , Camundongos , Panax/química , Extratos Vegetais/biossíntese , Extratos Vegetais/isolamento & purificação , Extratos Vegetais/farmacologia , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , beta-Glucosidase/genética
16.
Planta Med ; 83(9): 805-811, 2017 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-28095586

RESUMO

Chlamydiae are widely distributed pathogens of human populations, which can lead to serious reproductive and other health problems. In our search for novel antichlamydial metabolites from marine derived-microorganisms, one new (1) and two known (2, 3) dimeric indole derivatives were isolated from the sponge-derived actinomycete Rubrobacter radiotolerans. The chemical structures of these metabolites were elucidated by NMR spectroscopic data as well as CD calculations. All three metabolites suppressed chlamydial growth in a concentration-dependent manner. Among them, compound 1 exhibited the most effective antichlamydial activity with IC50 values of 46.6 ~ 96.4 µM in the production of infectious progeny. Compounds appeared to target the mid-stage of the chlamydial developmental cycle by interfering with reticular body replication, but not directly inactivating the infectious elementary body.


Assuntos
Actinobacteria/química , Antibacterianos/isolamento & purificação , Chlamydia trachomatis/efeitos dos fármacos , Indóis/farmacologia , Actinobacteria/isolamento & purificação , Animais , Antibacterianos/química , Antibacterianos/farmacologia , Antibacterianos/toxicidade , Chlamydia trachomatis/fisiologia , Células HeLa , Humanos , Indóis/química , Indóis/isolamento & purificação , Indóis/toxicidade , Estrutura Molecular , Petrosia/microbiologia
17.
Microb Ecol ; 73(2): 338-352, 2017 02.
Artigo em Inglês | MEDLINE | ID: mdl-27614749

RESUMO

Marine Actinobacteria are emerging as an unexplored source for natural product discovery. Eighty-seven deep-sea coral reef invertebrates were collected during an oceanographic expedition at the submarine Avilés Canyon (Asturias, Spain) in a range of 1500 to 4700 m depth. From these, 18 cultivable bioactive Actinobacteria were isolated, mainly from corals, phylum Cnidaria, and some specimens of phyla Echinodermata, Porifera, Annelida, Arthropoda, Mollusca and Sipuncula. As determined by 16S rRNA sequencing and phylogenetic analyses, all isolates belong to the phylum Actinobacteria, mainly to the Streptomyces genus and also to Micromonospora, Pseudonocardia and Myceligenerans. Production of bioactive compounds of pharmacological interest was investigated by high-performance liquid chromatography (HPLC) and gas chromatography-mass spectrometry (GC-MS) techniques and subsequent database comparison. Results reveal that deep-sea isolated Actinobacteria display a wide repertoire of secondary metabolite production with a high chemical diversity. Most identified products (both diffusible and volatiles) are known by their contrasted antibiotic or antitumor activities. Bioassays with ethyl acetate extracts from isolates displayed strong antibiotic activities against a panel of important resistant clinical pathogens, including Gram-positive and Gram-negative bacteria, as well as fungi, all of them isolated at two main hospitals (HUCA and Cabueñes) from the same geographical region. The identity of the active extracts components of these producing Actinobacteria is currently being investigated, given its potential for the discovery of pharmaceuticals and other products of biotechnological interest.


Assuntos
Actinobacteria/química , Actinobacteria/classificação , Actinobacteria/isolamento & purificação , Antozoários/microbiologia , Produtos Biológicos/farmacologia , Filogenia , Actinobacteria/genética , Animais , Antibacterianos/farmacologia , Antineoplásicos/farmacologia , Bactérias/efeitos dos fármacos , Sequência de Bases , Biodiversidade , Produtos Biológicos/química , Produtos Biológicos/isolamento & purificação , Bioprospecção , Linhagem Celular Tumoral/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Cromatografia Líquida de Alta Pressão , Classificação , Recifes de Corais , DNA Bacteriano , Ecossistema , Cromatografia Gasosa-Espectrometria de Massas , Genes Bacterianos , Invertebrados/microbiologia , Biologia Marinha , Extratos Vegetais , RNA Ribossômico 16S/genética , Água do Mar , Metabolismo Secundário , Espanha , Streptomyces/classificação , Streptomyces/isolamento & purificação
18.
Nat Prod Commun ; 11(3): 401-4, 2016 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-27169191

RESUMO

Analysis of an antimicrobial extract prepared from culture broth of the marine-derived actinomycete Nocardiopsis sp. (strain G057) led to the isolation of twelve compounds, 1-12. Compound 1 (2-[(2R-hydroxypropanoyl)amino]benzamide) was found to be a new enantiomeric isomer while compounds 2 (3-acetyl-4-hydroxycinnoline) and 3 (3,3'-bis-indole) were isolated from a natural source for the first time. The structures of 1-12 were determined by analyses of MS and 2D NMR data. All compounds were evaluated for their antimicrobial activity against a panel of clinically significant microorganisms. Compound 1 selectively inhibited Escherichia coli (MIC: 16 µg/mL). Compounds 2 and 3 exhibited antimicrobial activity against several strains of both Gram-positive and Gram-negative bacteria, and the yeast Candida albicans. Cytotoxic evaluation of compounds 1-3 against four cancer cell lines (KB, LU-1, HepG-2 and MCF-7) indicated that compound 3 produced a weak inhibition against KB and LU cell lines. Two remaining compounds, 1 and 2 were not cytotoxic, even at the concentration of 128 µg/mL.


Assuntos
Actinobacteria/metabolismo , Antibacterianos/farmacologia , Antifúngicos/farmacologia , Antineoplásicos/farmacologia , Actinobacteria/química , Antibacterianos/química , Antifúngicos/química , Antineoplásicos/química , Estrutura Molecular , Oceanos e Mares , Água do Mar/microbiologia , Vietnã , Microbiologia da Água
19.
Chem Biodivers ; 13(6): 727-36, 2016 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-27128202

RESUMO

Endophytic actinobacteria from the Brazilian medicinal plant Lychnophora ericoides were isolated for the first time, and the biological potential of their secondary metabolites was evaluated. A phylogenic analysis of isolated actinobacteria was accomplished with 16S rRNA gene sequencing, and the predominance of the genus Streptomyces was observed. All strains were cultured on solid rice medium, and ethanol extracts were evaluated with antimicrobial and cytotoxic assays against cancer cell lines. As a result, 92% of the extracts showed a high or moderate activity against at least one pathogenic microbial strain or cancer cell line. Based on the biological and chemical analyses of crude extracts, three endophytic strains were selected for further investigation of their chemical profiles. Sixteen compounds were isolated, and 3-hydroxy-4-methoxybenzamide (9) and 2,3-dihydro-2,2-dimethyl-4(1H)-quinazolinone (15) are reported as natural products for the first time in this study. The biological activity of the pure compounds was also assessed. Compound 15 displayed potent cytotoxic activity against all four tested cancer cell lines. Nocardamine (2) was only moderately active against two cancer cell lines but showed strong activity against Trypanosoma cruzi. Our results show that endophytic actinobacteria from L. ericoides are a promising source of bioactive compounds.


Assuntos
Actinobacteria/isolamento & purificação , Actinobacteria/metabolismo , Antineoplásicos Fitogênicos/farmacologia , Antiprotozoários/farmacologia , Asteraceae/microbiologia , Produtos Biológicos/farmacologia , Metabolismo Secundário , Actinobacteria/química , Antineoplásicos Fitogênicos/química , Antineoplásicos Fitogênicos/isolamento & purificação , Antiprotozoários/química , Antiprotozoários/isolamento & purificação , Produtos Biológicos/química , Produtos Biológicos/isolamento & purificação , Brasil , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Relação Dose-Resposta a Droga , Ensaios de Seleção de Medicamentos Antitumorais , Humanos , Estrutura Molecular , Testes de Sensibilidade Parasitária , Plantas Medicinais/microbiologia , Relação Estrutura-Atividade , Trypanosoma cruzi/efeitos dos fármacos
20.
Recent Pat Biotechnol ; 9(2): 116-29, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-26961669

RESUMO

BACKGROUND: Hexavalent chromium (Cr(6+)) contamination is one of the mejor problems of environmental protection for its carcinogenic effect on human health. Remediation of Cr(6+) contaminated environment thus becomes highest priority. METHODS: A Gram positive tiny rod shaped chromate (Cr(6+)) reducing bacterium strain A1 was isolated from uranium ore collected from Jaduguda, East Singhbhum, Jharkhand. The strain was identified and characterized in terms of its potential to reduce more toxic Cr(6+) to its less toxic form for its application in bioremediation of Cr(6+) contaminated environments. RESULTS: 16S rRNA gene based phylogentic analysis identified the strain as Microbacterium oleivorans. Along with Cr(6+), the strain showed resistance to other heavy metals including Ag, Cu, Co, Hg, Ni and Zn also. Complete reduction of Cr(6+) (750 µM) was achieved within 84 h with optimum reduction at pH 9 and 30°C. Effect of different parameters including cell mass concentration, pH, induction with Cr(6+), SO4 = ion, heavy metals, etc. on Cr(6+) reduction were studied thoroughly at resting cell condition to study its potential towards Cr(6+) bioremediation. Chromate reductase gene (chrA) was detected within this strain. CONCLUSION: Along with presence of appropriate genetic determinant, efficient Cr(6+) reduction ability of the strain indicated its potential for developing redox based Cr(6+) remediation system for varied concentrations of Cr6+ under a wide range of environmental conditions. Patent data have suggested the efficient application of Cr(6+) reducing bacteria in cleaning up of Cr(6+) contaminated environments.


Assuntos
Actinobacteria/química , Biodegradação Ambiental , Patentes como Assunto , Actinobacteria/genética , Cromo/química , Humanos , Oxirredução , Oxirredutases/química , Oxirredutases/genética , Filogenia , RNA Ribossômico 16S/genética , Urânio/química
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