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1.
Microb Pathog ; 193: 106758, 2024 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-38906493

RESUMO

The present study explores the bioinspired green synthesis of zinc oxide nanoparticles (ZnONPs) using marine Streptomyces plicatus and its potent antibacterial, antibiofilm activity against dental caries forming Streptococcus mutans MTCC and S. mutans clinical isolate (CI), cytotoxicity against oral KB cancer cells, hemolysis against blood erythrocytes and artemia toxicity. The bioinspired ZnONPs showed a distinctive absorption peak at 375 nm in UV-Vis spectra, the FT-IR spectra divulged the active functional groups, and XRD confirmed the crystalline nature of the nanoparticles with an average grain size of 41.76 nm. SEM analysis evidenced hexagonal morphology, and EDX spectra affirmed the presence of zinc. The ZnONPs exerted higher antagonistic activity against S. mutans MTCC (Inhibitory zone: 19 mm; MIC: 75 µg/ml) than S. mutans CI (Inhibitory zone: 17 mm; MIC: 100 µg/ml). Results of biofilm inhibitory activity showed a concentration-dependent reduction with S. mutans MTCC (15 %-95 %) more sensitive than S. mutans CI (13 %-89 %). The 50 % biofilm inhibitory concentration (BIC50) of ZnONPs against S. mutans MTCC was considerably lower (71.76 µg/ml) than S. mutans CI (78.13 µg/ml). Confocal Laser Scanning Microscopic visuals clearly implied that ZnONPs effectively distorted the biofilm architecture of both S. mutans MTCC and S. mutans CI. This was further bolstered by a remarkable rise in protein leakage (19 %-85 %; 15 %-77 %) and a fall in exopolysaccharide production (34 mg-7 mg; 49 mg-12 mg). MTT cytotoxicity of ZnONPs recorded an IC50 value of 22.06 µg/ml against KB cells. Acridine orange/ethidium bromide staining showed an increasing incidence of apoptosis in KB cells. Brine shrimp cytotoxicity using Artemia salina larvae recorded an LC50 value of 78.41 µg/ml. Hemolysis assay substantiated the biocompatibility of the ZnONPs. This study underscores the multifaceted application of bioinspired ZnONPs in dentistry.


Assuntos
Antibacterianos , Artemia , Biofilmes , Hemólise , Testes de Sensibilidade Microbiana , Streptococcus mutans , Streptomyces , Óxido de Zinco , Streptomyces/química , Streptomyces/metabolismo , Óxido de Zinco/farmacologia , Óxido de Zinco/química , Biofilmes/efeitos dos fármacos , Animais , Antibacterianos/farmacologia , Antibacterianos/química , Artemia/efeitos dos fármacos , Streptococcus mutans/efeitos dos fármacos , Humanos , Hemólise/efeitos dos fármacos , Eritrócitos/efeitos dos fármacos , Nanopartículas/química , Química Verde , Espectroscopia de Infravermelho com Transformada de Fourier , Linhagem Celular Tumoral , Organismos Aquáticos/química , Difração de Raios X
2.
Microb Cell Fact ; 23(1): 181, 2024 Jun 18.
Artigo em Inglês | MEDLINE | ID: mdl-38890640

RESUMO

BACKGROUND: Volatile compounds are key elements in the interaction and communication between organisms at both interspecific and intraspecific levels. In complex bacterial communities, the emission of these fast-acting chemical messengers allows an exchange of information even at a certain distance that can cause different types of responses in the receiving organisms. The changes in secondary metabolism as a consequence of this interaction arouse great interest in the field of searching for bioactive compounds since they can be used as a tool to activate silenced metabolic pathways. Regarding the great metabolic potential that the Actinobacteria group presents in the production of compounds with attractive properties, we evaluated the reply the emitted volatile compounds can generate in other individuals of the same group. RESULTS: We recently reported that volatile compounds released by different streptomycete species trigger the modulation of biosynthetic gene clusters in Streptomyces spp. which finally leads to the activation/repression of the production of secondary metabolites in the recipient strains. Here we present the application of this rationale in a broader bacterial community to evaluate volatiles as signaling effectors that drive the activation of biosynthesis of bioactive compounds in other members of the Actinobacteria group. Using cocultures of different actinobacteria (where only the volatile compounds reach the recipient strain) we were able to modify the bacterial secondary metabolism that drives overproduction (e.g., granaticins, actiphenol, chromomycins) and/or de novo production (e.g., collismycins, skyllamycins, cosmomycins) of compounds belonging to different chemical species that present important biological activities. CONCLUSIONS: This work shows how the secondary metabolism of different Actinobacteria species can vary significantly when exposed in co-culture to the volatile compounds of other phylum-shared bacteria, these effects being variable depending on strains and culture media. This approach can be applied to the field of new drug discovery to increase the battery of bioactive compounds produced by bacteria that can potentially be used in treatments for humans and animals.


Assuntos
Actinobacteria , Metabolismo Secundário , Compostos Orgânicos Voláteis , Actinobacteria/metabolismo , Actinobacteria/genética , Compostos Orgânicos Voláteis/metabolismo , Streptomyces/metabolismo , Streptomyces/genética , Família Multigênica
3.
Environ Res ; 251(Pt 1): 118666, 2024 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-38462087

RESUMO

In the present scenario, food security is of major concern due to exponentially increasing population and depleted crop production. The fungal diseases have contributed majorly to the scarcity of staple food products and economic loss worldwide. This problem could be tackled by preventing the crop loss during both pre and post-harvest seasons. During the current investigation, the bioactive compound eicosane was extracted from Streptomyces sp. KF15, subjected to purification and identified based on mass spectrometry and NMR analysis. The evaluation of in-vitro antifungal activity was done by poisoned food method, SEM analysis and growth pattern analysis. The bioactive compound eicosane with molecular weight of 282.5475 g/mol was purified by column chromatography and the straight chain hydrocarbon structure of CH3CH2(18)CH3 was elucidated by NMR analysis. In poisoned food assay, eicosane effectively inhibited the radial growth of all tested fungal pathogens; F. oxysporum was found to be the most sensitive with 24.2%, 33.3%, 42.4%, and 63.6% inhibition at 25-100 µg/ml concentrations. The SEM micrograph established clear differences in the morphology of eicosane treated fungi with damaged hyphae, flaccid mycelium and collapsed spores as compared to the tubular, turgid and entire fungi in control sample. Finally, the growth curve assay depicted the right side shift in the pattern of eicosane treated fungi indicating the delay in adapting to the conditions of growth and multiplication. The findings of this study encourage further research and development towards the novel antifungal drugs that can act against major phytopathogens.


Assuntos
Antifúngicos , Streptomyces , Streptomyces/química , Antifúngicos/farmacologia , Antifúngicos/química , Produtos Agrícolas/microbiologia , Fungicidas Industriais/farmacologia , Doenças das Plantas/prevenção & controle , Doenças das Plantas/microbiologia , Fungos/efeitos dos fármacos
4.
Appl Microbiol Biotechnol ; 108(1): 49, 2024 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-38183485

RESUMO

Apple fire blight, caused by the bacterium Erwinia amylovora, is a devastating disease of apple and pear trees. Biological control methods have attracted much attention from researchers to manage plant diseases as they are eco-friendly and viable alternatives to synthetic pesticides. Herein, we isolated Streptomyces sp. JCK-8055 from the root of pepper and investigated its mechanisms of action against E. amylovora. Streptomyces sp. JCK-8055 produced aureothricin and thiolutin, which antagonistically affect E. amylovora. JCK-8055 and its two active metabolites have a broad-spectrum in vitro activity against various phytopathogenic bacteria and fungi. They also effectively suppressed tomato bacterial wilt and apple fire blight in in vivo experiments. Interestingly, JCK-8055 colonizes roots as a tomato seed coating and induces apple leaf shedding at the abscission zone, ultimately halting the growth of pathogenic bacteria. Additionally, JCK-8055 can produce the plant growth regulation hormone indole-3-acetic acid (IAA) and hydrolytic enzymes, including protease, gelatinase, and cellulase. JCK-8055 treatment also triggered the expression of salicylate (SA) and jasmonate (JA) signaling pathway marker genes, such as PR1, PR2, and PR3. Overall, our findings demonstrate that Streptomyces sp. JCK-8055 can control a wide range of plant diseases, particularly apple fire blight, through a combination of mechanisms such as antibiosis and induced resistance, highlighting its excellent potential as a biocontrol agent. KEY POINTS: • JCK-8055 produces the systemic antimicrobial metabolites, aureothricin, and thiolutin. • JCK-8055 treatment upregulates PR gene expression in apple plants against E. amylovora. • JCK-8055 controls plant diseases with antibiotics and induced resistance.


Assuntos
Malus , Pirróis , Compostos de Sulfidrila , Endopeptidases
5.
Lett Appl Microbiol ; 77(9)2024 Sep 02.
Artigo em Inglês | MEDLINE | ID: mdl-39264087

RESUMO

This study reports the isolation and characterization of a Streptomyces sp. from soil, capable of producing bioactive secondary metabolites active against a variety of bacterial human pathogens. We targeted the antimicrobial activity against Escherichia coli ATCC-BAA 2469, a clinically relevant strain of bacteria harbouring resistance genes for carbapenems, extended spectrum beta-lactams, tetracyclines, fluoroquinones, etc. Preliminary screening using the spot inoculation technique identified Streptomyces sp. NP73 as the potent strain among the 74 isolated Actinomycetia strain. 16S rRNA gene and whole genome sequencing (WGS) confirmed its taxonomical identity and helped in the construction of the phylogenetic tree. WGS revealed the predicted pathways and biosynthetic gene clusters responsible for producing various types of antibiotics including the isolated compound. Bioactivity guided fractionation and chemical characterization of the active fraction, carried out using liquid chromatography, gas chromatography-mass spectrometry, infra-red spectroscopy, and nuclear magnetic resonance spectroscopy, led to the tentative identification of the active compound as Pyrrolo[1,2-a] pyrazine-1,4-dione, hexahydro-, a diketopiperazine molecule. This compound exhibited excellent antimicrobial and anti-biofilm properties against E. coli ATCC-BAA 2469 with an MIC value of 15.64 µg ml-1, and the low cytotoxicity of the compound identified in this study provides hope for future drug development.


Assuntos
Antibacterianos , Farmacorresistência Bacteriana Múltipla , Escherichia coli , Testes de Sensibilidade Microbiana , Filogenia , RNA Ribossômico 16S , Microbiologia do Solo , Streptomyces , Streptomyces/química , Streptomyces/isolamento & purificação , Streptomyces/genética , Streptomyces/classificação , Streptomyces/metabolismo , Escherichia coli/efeitos dos fármacos , Escherichia coli/genética , Índia , Antibacterianos/farmacologia , Antibacterianos/química , Antibacterianos/isolamento & purificação , RNA Ribossômico 16S/genética , Florestas , Biofilmes/efeitos dos fármacos , Sequenciamento Completo do Genoma , Humanos , Família Multigênica
6.
Mar Drugs ; 22(7)2024 Jul 09.
Artigo em Inglês | MEDLINE | ID: mdl-39057421

RESUMO

A bioassay-guided chemical investigation of a bacterium, Streptomyces sp. CMB-MRB032, isolated from sheep feces collected near Bathurst, Victoria, Australia, yielded the known polyketide antimycins A4a (1) and A2a (2) as potent inhibitors of Dirofilaria immitis (heartworm) microfilaria (mf) motility (EC50 0.0013-0.0021 µg/mL), along with the octapeptide surugamide A (3) and the new N-methylated analog surugamide K (4). With biological data suggesting surugamides may also exhibit activity against D. immitis, a GNPS molecular network analysis of a library of microbes sourced from geographically diverse Australian ecosystems identified a further five taxonomically and chemically distinct surugamide producers. Scaled-up cultivation of one such producer, Streptomyces sp. CMB-M0112 isolated from a marine sediment collected at Shorncliff, Qld, Australia, yielded 3 along with the new acyl-surugamides A1-A4 (5-8). Solid-phase peptide synthesis provided additional synthetic analogs, surugamides S1-S3 (9-11), while derivatization of 3 returned the semi-synthetic surugamide S4 (12) and acyl-surugamides AS1-AS3 (13-15). The natural acyl-surugamide A3 (7) and semi-synthetic acyl-surugamide AS3 (15) were shown to selectively inhibit D. immitis mf motility (EC50 3.3-3.4 µg/mL), however, unlike antimycins 1 and 2, were inactive against the gastrointestinal nematode Haemonchus contortus L1-L3 larvae (EC50 > 25 µg/mL) and were not cytotoxic to mammalian cells (human colorectal carcinoma SW620, IC50 > 30 µg/mL). A structure-activity relationship (SAR) study on the surugamides 3-15 revealed that selective acylation of the Lys3-ε-NH2 correlates with anthelmintic activity.


Assuntos
Dirofilaria immitis , Streptomyces , Animais , Streptomyces/química , Dirofilaria immitis/efeitos dos fármacos , Austrália , Ovinos , Fezes/parasitologia , Fezes/microbiologia
7.
Mar Drugs ; 22(4)2024 Apr 17.
Artigo em Inglês | MEDLINE | ID: mdl-38667798

RESUMO

Three pairs of enantiomers (1-3)-the new 12R-aloesol (1a) and two new fatty acids (2 and 3)-and one new natural product (4) together three known compounds (5-7) were isolated from a coral-reef-derived Streptomyces sp. SCSIO 66814. Their structures were determined through extensive spectroscopic analysis, chiral analysis, and single-crystal X-ray diffraction data. Compounds 2 and 3 were presumed to be intermediates for further generating homononactic acid (5) and nonactic acid, and the latter two molecules were able to act as precursors to form macrotetrolides with remarkable biological activity. The isolation of related precursors, compounds 2-5, provided more evidence to support the proposal of a plausible biosynthetic pathway for nonactic acid and its homologs. Additionally, (+)-1 exhibited a weak activity against DPPH radicals.


Assuntos
Antozoários , Cromonas , Streptomyces , Streptomyces/metabolismo , Streptomyces/química , Cromonas/química , Cromonas/isolamento & purificação , Cromonas/farmacologia , Estereoisomerismo , Antozoários/química , Animais , Cristalografia por Raios X , Ácidos Graxos/química , Ácidos Graxos/isolamento & purificação , Produtos Biológicos/química , Produtos Biológicos/farmacologia , Produtos Biológicos/isolamento & purificação , Estrutura Molecular
8.
Mar Drugs ; 22(5)2024 Apr 25.
Artigo em Inglês | MEDLINE | ID: mdl-38786587

RESUMO

Marine symbiotic and epiphyte microorganisms are sources of bioactive or structurally novel natural products. Metabolic blockade-based genome mining has been proven to be an effective strategy to accelerate the discovery of natural products from both terrestrial and marine microorganisms. Here, the metabolic blockade-based genome mining strategy was applied to the discovery of other metabolites in a sea anemone-associated Streptomyces sp. S1502. We constructed a mutant Streptomyces sp. S1502/Δstp1 that switched to producing the atypical angucyclines WS-5995 A-E, among which WS-5995 E is a new compound. A biosynthetic gene cluster (wsm) of the angucyclines was identified through gene knock-out and heterologous expression studies. The biosynthetic pathways of WS-5995 A-E were proposed, the roles of some tailoring and regulatory genes were investigated, and the biological activities of WS-5995 A-E were evaluated. WS-5995 A has significant anti-Eimeria tenell activity with an IC50 value of 2.21 µM. The production of antibacterial streptopyrroles and anticoccidial WS-5995 A-E may play a protective role in the mutual relationship between Streptomyces sp. S1502 and its host.


Assuntos
Família Multigênica , Anêmonas-do-Mar , Streptomyces , Streptomyces/genética , Streptomyces/metabolismo , Animais , Antibacterianos/farmacologia , Vias Biossintéticas/genética , Genoma Bacteriano , Produtos Biológicos/farmacologia , Antraquinonas/farmacologia , Anguciclinas e Anguciclinonas
9.
Mar Drugs ; 22(1)2024 Jan 11.
Artigo em Inglês | MEDLINE | ID: mdl-38248664

RESUMO

This study aims to explore the potential inhibition effects of staurosporine isolated from a Streptomyces sp. SNC087 strain obtained from seawater on nasal polyps. Staurosporine possesses antimicrobial and antihypertensive activities. This research focuses on investigating the effects of staurosporine on suppressing the growth and development of nasal polyps and elucidating the underlying mechanisms involved. The experimental design includes in vitro and ex vivo evaluations to assess the inhibition activity and therapeutic potential of staurosporine against nasal polyps. Nasal polyp-derived fibroblasts (NPDFs) were stimulated with TGF-ß1 in the presence of staurosporine. The levels of α-smooth muscle actin (α-SMA), collagen type-I (Col-1), fibronectin, and phosphorylated (p)-Smad 2 were investigated using Western blotting. VEGF expression levels were analyzed in nasal polyp organ cultures treated with staurosporine. TGF-ß1 stimulated the production of Col-1, fibronectin, and α-SMA and was attenuated by staurosporine pretreatment. Furthermore, these inhibitory effects were mediated by modulation of the signaling pathway of Smad 2 in TGF-ß1-induced NPDFs. Staurosporine also inhibits the production of VEGF in ex vivo NP tissues. The findings from this study will contribute to a better understanding of staurosporine's role in nasal polyp management and provide insights into its mechanisms of action.


Assuntos
Pólipos Nasais , Streptomyces , Humanos , Fibronectinas , Pólipos Nasais/tratamento farmacológico , Estaurosporina/farmacologia , Fator de Crescimento Transformador beta1 , Fator A de Crescimento do Endotélio Vascular
10.
Mar Drugs ; 22(4)2024 Apr 21.
Artigo em Inglês | MEDLINE | ID: mdl-38667806

RESUMO

Polyene macrolactams are a special group of natural products with great diversity, unique structural features, and a wide range of biological activities. Herein, a cryptic gene cluster for the biosynthesis of putative macrolactams was disclosed from a sponge-associated bacterium, Streptomyces sp. DSS69, by genome mining. Cloning and heterologous expression of the whole biosynthetic gene cluster led to the discovery of weddellamycin, a polyene macrolactam bearing a 23/5/6 ring skeleton. A negative regulator, WdlO, and two positive regulators, WdlA and WdlB, involved in the regulation of weddellamycin production were unraveled. The fermentation titer of weddellamycin was significantly improved by overexpression of wdlA and wdlB and deletion of wdlO. Notably, weddellamycin showed remarkable antibacterial activity against various Gram-positive bacteria including MRSA, with MIC values of 0.10-0.83 µg/mL, and antifungal activity against Candida albicans, with an MIC value of 3.33 µg/mL. Weddellamycin also displayed cytotoxicity against several cancer cell lines, with IC50 values ranging from 2.07 to 11.50 µM.


Assuntos
Antibacterianos , Lactamas Macrocíclicas , Testes de Sensibilidade Microbiana , Família Multigênica , Streptomyces , Streptomyces/genética , Streptomyces/metabolismo , Antibacterianos/farmacologia , Antibacterianos/biossíntese , Antibacterianos/química , Humanos , Lactamas Macrocíclicas/farmacologia , Lactamas Macrocíclicas/química , Lactamas Macrocíclicas/isolamento & purificação , Polienos/farmacologia , Polienos/isolamento & purificação , Polienos/química , Candida albicans/efeitos dos fármacos , Linhagem Celular Tumoral , Regiões Antárticas , Animais , Poríferos/microbiologia , Antifúngicos/farmacologia , Antifúngicos/química , Antifúngicos/isolamento & purificação
11.
Chem Biodivers ; : e202401154, 2024 Jul 14.
Artigo em Inglês | MEDLINE | ID: mdl-39003590

RESUMO

A novel compound streptothiomycin F (1), and a new natural product, N-(5-nitropentyl)acetamide (2), were discovered alongside ten previously identified compounds (3-12) through solid fermentation of marine-derived Streptomyces sp. ZS-A31 based on rice. The chemical structures of compounds 1-2 were elucidated using 1D and 2D NMR, as well as HRESIMS data analysis. Evaluation of all isolated compounds for their antibiofilm and antibacterial activities against P. aeruginosa was carried out using microdilution and crystal violet staining methods. Results highlighted the weak potency of the known compounds lumichrome (3) and vanillic acid (7) in inhibiting biofilm formation.

12.
Chem Biodivers ; 21(5): e202400090, 2024 May.
Artigo em Inglês | MEDLINE | ID: mdl-38486477

RESUMO

Streptomide (1), a new amide analogue, streptomynone (2), a new quinolinone, and ten known compounds including three aliphatic acids (3-5), two amides (6-7), four cyclic dipeptides (8-11), and an adenosine (12) were isolated from the fermentation broth of Streptomyces sp. YIM S01983 isolated from a sediment sample collected in Bendong Village, Huadong Town, Chuxiong, China. Their structures were determined by analysis of the 1D/2D-NMR and HR-ESI-MS spectra. Compound 12 presented weak antimicrobial activities against Candida albicans and Aligenes faecalis (MIC=64 µg/mL). Compounds 7 and 12 showed weak cytotoxic activity against MHCC97H.


Assuntos
Amidas , Candida albicans , Testes de Sensibilidade Microbiana , Quinolonas , Streptomyces , Streptomyces/química , Streptomyces/metabolismo , Amidas/química , Amidas/farmacologia , Amidas/isolamento & purificação , Candida albicans/efeitos dos fármacos , Quinolonas/química , Quinolonas/farmacologia , Quinolonas/isolamento & purificação , Humanos , Linhagem Celular Tumoral , Antibacterianos/farmacologia , Antibacterianos/química , Antibacterianos/isolamento & purificação , Antifúngicos/farmacologia , Antifúngicos/química , Antifúngicos/isolamento & purificação , Antineoplásicos/farmacologia , Antineoplásicos/química , Antineoplásicos/isolamento & purificação , Enterococcus faecalis/efeitos dos fármacos , Estrutura Molecular , Relação Estrutura-Atividade , Ensaios de Seleção de Medicamentos Antitumorais
13.
Molecules ; 29(9)2024 Apr 28.
Artigo em Inglês | MEDLINE | ID: mdl-38731531

RESUMO

Actinomycetes have long been recognized as an important source of antibacterial natural products. In recent years, actinomycetes in extreme environments have become one of the main research directions. Streptomyces sp. KN37 was isolated from the cold region of Kanas in Xinjiang. It demonstrated potent antimicrobial activity, but the primary active compounds remained unclear. Therefore, we aimed to combine genomics with traditional isolation methods to obtain bioactive compounds from the strain KN37. Whole-genome sequencing and KEGG enrichment analysis indicated that KN37 possesses the potential for synthesizing secondary metabolites, and 41 biosynthetic gene clusters were predicted, some of which showed high similarity to known gene clusters responsible for the biosynthesis of antimicrobial antibiotics. The traditional isolation methods and activity-guided fractionation were employed to isolate and purify seven compounds with strong bioactivity from the fermentation broth of the strain KN37. These compounds were identified as 4-(Diethylamino)salicylaldehyde (1), 4-Nitrosodiphenylamine (2), N-(2,4-Dimethylphenyl)formamide (3), 4-Nitrocatechol (4), Methylsuccinic acid (5), Phenyllactic acid (6) and 5,6-Dimethylbenzimidazole (7). Moreover, 4-(Diethylamino)salicylaldehyde exhibited the most potent inhibitory effect against Rhizoctonia solani, with an EC50 value of 14.487 mg/L, while 4-Nitrosodiphenylamine showed great antibacterial activity against Erwinia amylovora, with an EC50 value of 5.715 mg/L. This study successfully isolated several highly active antimicrobial compounds from the metabolites of the strain KN37, which could contribute as scaffolds for subsequent chemical synthesis. On the other hand, the newly predicted antibiotic-like substances have not yet been isolated, but they still hold significant research value. They are instructive in the study of active natural product biosynthetic pathways, activation of silent gene clusters, and engineering bacteria construction.


Assuntos
Genômica , Família Multigênica , Streptomyces , Streptomyces/genética , Streptomyces/metabolismo , Streptomyces/química , Genômica/métodos , Antibacterianos/farmacologia , Antibacterianos/química , Antibacterianos/isolamento & purificação , Antibacterianos/biossíntese , Testes de Sensibilidade Microbiana , Produtos Biológicos/farmacologia , Produtos Biológicos/química , Produtos Biológicos/isolamento & purificação , Anti-Infecciosos/farmacologia , Anti-Infecciosos/química , Anti-Infecciosos/isolamento & purificação , Agricultura/métodos , Sequenciamento Completo do Genoma
14.
Molecules ; 29(7)2024 Mar 24.
Artigo em Inglês | MEDLINE | ID: mdl-38611738

RESUMO

The Streptomyces strain G222, isolated from a Vietnamese marine sediment, was confidently identified by 16S rRNA gene sequencing. Its AcOEt crude extract was successfully analyzed using non-targeted LC-MS/MS analysis, and molecular networking, leading to a putative annotation of its chemical diversity thanks to spectral libraries from GNPS and in silico metabolite structure prediction obtained from SIRIUS combined with the bioinformatics tool conCISE (Consensus Annotation Propagation of in silico Elucidations). This dereplication strategy allowed the identification of an interesting cluster of a series of putative cyclic and linear lipopeptides of the lichenysin and surfactin families. Lichenysins (3-7) were isolated from the sub-fraction, which showed significant anti-biofilm activity against Pseudomonas aeruginosa MUC-N1. Their structures were confirmed by detailed 1D and 2D NMR spectroscopy (COSY, HSQC, HMBC, TOCSY, ROESY) recorded in CD3OH, and their absolute configurations were determined using the modified Marfey's method. The isolated lichenysins showed anti-biofilm activity at a minimum concentration of 100 µM. When evaluated for antibacterial activity against a panel of Gram-positive and Gram-negative strains, two isolated lichenysins exhibited selective activity against the MRSA strain without affecting its growth curve and without membranotropic activity. This study highlights the power of the MS/MS spectral similarity strategy using computational methods to obtain a cross-validation of the annotated molecules from the complex metabolic profile of a marine sediment-derived Streptomyces extract. This work provides the first report from a Streptomyces strain of combined cyclic and linear lichenysins and surfactins, known to be characteristic compounds of the genus Bacillus.


Assuntos
Sedimentos Geológicos , Espectrometria de Massas em Tandem , Humanos , Cromatografia Líquida , RNA Ribossômico 16S , Vietnã
15.
New Phytol ; 238(2): 765-780, 2023 04.
Artigo em Inglês | MEDLINE | ID: mdl-36653958

RESUMO

Metabolites including antibiotics, enzymes, and volatiles produced by plant-associated bacteria are key factors in plant-microbiota interaction that regulates various plant biological processes. There should be crucial mediators responsible for their entry into host plants. However, less is known about the identities of these plant transporters. We report that the Arabidopsis Nitrate Transporter1 (NRT1)/NPF protein NPF2.13 functions in plant uptake of tunicamycin (TM), a natural antibiotic produced by several Streptomyces spp., which inhibits protein N-glycosylation. Loss of NPF2.13 function resulted in enhanced TM tolerance, whereas NPF2.13 overexpression led to TM hypersensitivity. Transport assays confirmed that NPF2.13 is a H+ /TM symporter and the transport is not affected by other substrates like nitrate. NPF2.13 exclusively showed TM transport activity among tested NPFs. Tunicamycin uptake from TM-producing Streptomyces upregulated the expression of nitrate-related genes including NPF2.13. Moreover, nitrate allocation to younger leaves was promoted by TM in host plants. Tunicamycin could also benefit plant defense against the pathogen. Notably, the TM effects were significantly repressed in npf2.13 mutant. Overall, this study identifies NPF2.13 protein as an important TM transporter in plant-microbe interaction and provides insights into multiple facets of NPF proteins in modulating plant nutrition and defense by transporting exterior bacterial metabolites.


Assuntos
Proteínas de Arabidopsis , Arabidopsis , Arabidopsis/genética , Arabidopsis/metabolismo , Proteínas de Plantas/metabolismo , Tunicamicina/farmacologia , Nitratos/metabolismo , Proteínas de Transporte de Ânions/metabolismo , Proteínas de Arabidopsis/metabolismo , Regulação da Expressão Gênica de Plantas
16.
Arch Microbiol ; 205(6): 244, 2023 May 20.
Artigo em Inglês | MEDLINE | ID: mdl-37209150

RESUMO

Streptomyces sp. RS2 was isolated from an unidentified sponge collected around Randayan Island, Indonesia. The genome of Streptomyces sp. RS2 consists of a linear chromosome of 9,391,717 base pairs with 71.9% of G + C content, 8270 protein-coding genes, as well as 18 rRNA and 85 tRNA loci. Twenty-eight putative secondary metabolites biosynthetic gene clusters (BGCs) were identified in the genome sequence. Nine of them have 100% similarity to BGCs for albaflavenone, α-lipomycin, coelibactin, coelichelin, ectoine, geosmin, germicidin, hopene, and lanthionine (SapB). The remaining 19 BGCs have low (< 50%) or moderate (50-80%) similarity to other known secondary metabolite BGCs. Biological activity assays of extracts from 21 different cultures of the RS2 strain showed that SCB ASW was the best medium for the production of antimicrobial and cytotoxic compounds. Streptomyces sp. RS2 has great potential to be a producer of novel secondary metabolites, particularly those with antimicrobial and antitumor activities.


Assuntos
Anti-Infecciosos , Antineoplásicos , Streptomyces , Genoma Bacteriano , Anti-Infecciosos/metabolismo , Antineoplásicos/farmacologia , Antineoplásicos/metabolismo , Metabolismo Secundário/genética , Família Multigênica
17.
Artigo em Inglês | MEDLINE | ID: mdl-37319004

RESUMO

Two new actinobacteria, designated strains IBSBF 2807T and IBSBF 2953T, isolated from scab lesions on potato tubers grown in the southern Brazilian states of Rio Grande do Sul and Santa Catarina, respectively, were characterized and identified through a polyphasic approach. Phylogenetic analyses of 16S rRNA sequences revealed that these two strains belong to the genus Streptomyces. Multilocus sequence analysis using five concatenated genes, atpD, gyrB, recA, rpoB and trpB, allocated strains IBSBF 2807T and IBSBF 2953T in distinct branches of Streptomyces phytopathogenic strains. PCR-RFLP analysis of the atpD gene also confirmed that these strains differ from the type strains of Streptomyces associated with potato scab. The morphological, physiological and biochemical characterization, along with the overall genome-related index properties, indicated that these two strains could be distinguished from their closest phylogenetic relatives and each other. According to the data, IBSBF 2807T and IBSBF 2953T represent two new Streptomyces species related to potato scab. The proposed names for these strains are Streptomyces hilarionis sp. nov. (IBSBF 2807T=CBMAI 2674T=ICMP 24297T=MUM 22.66T) and Streptomyces hayashii sp. nov (IBSBF 2953T=CBMAI 2675T=ICMP 24301T=MUM 22.68T).


Assuntos
Solanum tuberosum , Streptomyces , Ácidos Graxos/química , Análise de Sequência de DNA , Solanum tuberosum/microbiologia , Brasil , Filogenia , RNA Ribossômico 16S/genética , DNA Bacteriano/genética , Técnicas de Tipagem Bacteriana , Composição de Bases
18.
Environ Res ; 231(Pt 3): 116212, 2023 08 15.
Artigo em Inglês | MEDLINE | ID: mdl-37244496

RESUMO

Anthracnose is a devastating disease caused by the fungus Colletotrichum lindemuthianum (CL) in Vigna radiata (L.) R. Wilczek (mung bean). In the present study, an eco-friendly approach to control anthracnose infection, growth promotion and enhancement of defense response in mung bean plants using endophytic actinomycetes was performed. Among the 24 actinomycetes isolates from the Cleome rutidosperma plant, the isolate SND-2 exhibited a broad spectrum of antagonistic activity with 63.27% of inhibition against CL in the dual culture method. Further, the isolate SND-2 was identified as Streptomyces sp. strain SND-2 (SND-2) through the 16S rRNA gene sequence. In-vitro screening of plant growth trials confirmed that SND-2 has the potential to produce indole acetic acid, hydrogen cyanide, ammonia, phosphate solubilization, and siderophore. The in-vivo biocontrol study was performed with exogenous application of wettable talcum-based formulation of SND-2 strain to mitigate CL infection in mung bean seedlings. The results displayed maximum seed germination, vigor index, increased growth parameters, and lowest disease severity (43.63 ± 0.73) in formulation treated and pathogen challenged mung bean plants. Further, the application of SND-2 formulation with pathogen witnessed increased cellular defense through the maximum accumulation of lignin, hydrogen peroxide and phenol deposition in mung bean leaves compared with control treatments. Biochemical defense response exhibited upregulation of antioxidant enzymes such as phenylalanine ammonia-lyase, ß-1,-3-Glucanase, and peroxidase enzymes activities with increased phenolic (3.64 ± 0.11 mg/g fresh weight) and flavonoid (1.14 ± 0.05 mg/g fresh weight) contents in comparison with other treatments at 0, 4, 12, 24, 36, and 72 h post pathogen inoculation. This study demonstrated that formulation of Streptomyces sp. strain SND-2 is a potential source as a suppressive agent and plant growth promoter in mung bean plants upon C. lindemuthianum infestation and witnesses the elevation in cellular and biochemical defense against anthracnose disease.


Assuntos
Fabaceae , Streptomyces , Vigna , Vigna/química , Vigna/genética , Streptomyces/genética , RNA Ribossômico 16S/genética , Fabaceae/genética , Antioxidantes
19.
Mar Drugs ; 21(6)2023 Jun 16.
Artigo em Inglês | MEDLINE | ID: mdl-37367686

RESUMO

Nine sesquiterpenes, including eight pentalenenes (1-8) and one bolinane derivative (9), were isolated from the culture broth of a marine-derived actinobacterium Streptomyces qinglanensis 213DD-006. Among them, 1, 4, 7, and 9 were new compounds. Their planar structures were determined by spectroscopic methods (HRMS, 1D, and 2D NMR), and the absolute configuration was established by biosynthesis consideration and electronic-circular-dichroism (ECD) calculations. All the isolated compounds were screened for their cytotoxicity against six solid and seven blood cancer cell lines. Compounds 4-6 and 8 showed a moderate activity against all of the tested solid cell lines, with GI50 values ranging from 1.97 to 3.46 µM.


Assuntos
Antineoplásicos , Sesquiterpenos , Streptomyces , Estrutura Molecular , Antineoplásicos/química , Streptomyces/química , Sesquiterpenos/química
20.
Mar Drugs ; 21(4)2023 Apr 14.
Artigo em Inglês | MEDLINE | ID: mdl-37103379

RESUMO

Six angucyclines including three unreported compounds (1-3) were isolated from Streptomyces sp. XS-16 by overexpressing the native global regulator of SCrp (cyclic AMP receptor). The structures were characterized based on nuclear magnetic resonance (NMR) and spectrometry analysis and assisted by electronic circular dichroism (ECD) calculations. All compounds were tested for their antitumor and antimicrobial activities, and compound 1 showed different inhibitory activities against various tumor cell lines with IC50 values ranging from 0.32 to 5.33 µM.


Assuntos
Antineoplásicos , Streptomyces , Antineoplásicos/química , Streptomyces/metabolismo , Linhagem Celular Tumoral , Espectroscopia de Ressonância Magnética , Estrutura Molecular
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