Shunt products of aminoansamycins from aas1 overexpressed mutant strain of Streptomyces sp. S35 / 中国天然药物

Constitutively expression of the pathway-specific activators is an effective method to activate silent gene clusters and improve natural product production. In this study, nine shunt products of aminoansamycins (1-9) were identified from a recombinant mutant strain S35-LAL by overexpressed the large-ATP-binding regulator of the LuxR family (LAL) gene aas1 in Streptomyces sp. S35. All the compounds showed no anti-microbial, anti-T3SS and cytotoxic activities.

A new polyketide from marine-derived Streptomyces sp.MDW-06 / 中国中药杂志

To isolate and identify secondary metabolites of marine-derived Streptomyces sp.MDW-06,the isolations and purifications of compounds were performed by means of column chromatography over silica gel. And their structures were elucidated through the spectroscopic analysis of MS,NMR and specific rotations. The bioactivities were assayed by paper diffusion and DPPH method. From the fermentation broth of marine-derived Streptomyces sp.MDW-06,five compounds( 1-5) were isolated and identified as streptopentanoic acid( 1),germicidin A( 2),germicidin B( 3),isogermicidin A( 4),isogermicidin B( 5) and oxohygrolidin( 6),respectively. Compound 1 is a new compound. Compound 1 shows DPPH radical scavenging activity with 36. 4% at 100 mg·L~(-1).

Secondary Metabolites from Streptomyces sp CSDX076 for the First Time / Metabolitos secundarios de Streptomyces SP CSDX076 por primera vez

ABSTRACT Objective: To investigate the secondary metabolites from the cultures of Streptomyces sp CSDX076. Methods: The compounds were isolated using column chromatography and RP-18 medium-pressure liquid chromatography. Their structures were elucidated by one-dimensional and two-dimensional nuclear magnetic resonance spectroscopic methods in combination with mass spectrometry experiments. Results: Four compounds were isolated from the cultures of Streptomyces sp CSDX076 and identified as aurantiamide benzoate, deoxytryptoquivaline, 2-acetyl-3,5-dihydroxyl-benzene acetic acid, and 2-acetyl-3,5-dihydroxyl-benzene ester. Conclusion: It was the first time that the four isolated compounds were obtained from the Streptomyces genus.

RESUMEN Objetivo: Investigar los metabolitos secundarios de los cultivos de Streptomyces sp CSDX076. Métodos: Los compuestos fueron aislados usando la cromatografía de columna y cromatografía líquida RP-18 de presión media. Sus estructuras fueron dilucidadas mediante métodos espectroscópicos de resonancia magnética nuclear unidimensional y bidimensional, combinados con experimentos de espectrometría de masa. Resultados: Cuatro compuestos de culturas de Streptomyces sp CSDX076 fueron aislados e identificados como benzoato de aurantiamida, deoxitriptoquivalina, ácido acético 2-acetil-3,5-dihidroxil-benzeno, y éster 2-acetil-3,5-dihidroxil-benzeno. Conclusión: Fue la primera vez que los cuatro compuestos aislados se obtuvieron del género Streptomyces.

Secondary metabolites of Streptomyces sp. A1693 / 中国中药杂志

By means of various chromatographic methods such as Sephadex LH-20，ODS，and semi-preparative HPLC，ten compounds were isolated from Streptomyces sp. A1693 and their structures were elucidated on the basis of spectroscopic data and physico-chemical methods. The compounds comprised 5 butenolides，2 diketopiperazines，and 3 antimycin antibiotics. The structures were identified as (5)-5-(11-hydroxymethyloctyl)furan-2(5)-one (1), (5)-5-(11-hydroxy-11-methylheptyl)furan-2(5)-one (2), (5)-5-(11-methyl-12-oxooctyl) furan-2(5)-one (3), (5)-5-(11-hydroxy-11-methyloctyl)furan-2(5)-one (4), (5)-5-(11-hydroxy-12-methyloctyl)furan-2(5)-one（5），cyclo-Phe-Val (6)，cyclo-Phe-Ile (7)，uranchimycin A (8)，uranchimycin B (9)，and deisovalerylblastomycin (10). Among them，1 was defined as a new compound. All the compounds didn't show the cytotoxic activity against A549 cell line (IC₅₀>50 mg·L⁻¹).

Control Efficacy of Streptomyces sp. A501 against Ginseng Damping-off and Its Antifungal Substance

Ginseng damping-off, caused by the fungal pathogens Rhizoctonia solani and Pythium sp., is a critical disease in ginseng seedling. In a continuing effort to find microorganisms with the potential of acting as a biocontrol agent against Rhizoctonia damping-off, we found that a Streptomyces sp. A501 showed significant antifungal activity against Rhizoctonia solani. In field experiment to test the efficacy of Streptomyces sp. A501 in controlling ginseng damping-off, the incidence of damping-off disease was meaningfully reduced when ginseng seeds were soaked in the culture broth of Streptomyces sp. A501 before sowing. To perform characterization of the antifungal compound, we isolated it from the culture broth of strain A501 through Diaion HP-20 and silica gel column chromatographies and preparative high-performance liquid chromatography. The structure of the antifungal compound was assigned as fungichromin by spectroscopic methods, mainly nuclear magnetic resonance and electrospray ionization-mass analysis.

A new benzamide derivative from rice fermentation of Streptomyces sp. CPCC 202950 / 中国中药杂志

Eight compounds were isolated from the rice fermentation of Streptomyces sp. CPCC 202950 by a combination of various chromatographic techniques including column chromatography over silica, Sephadex LH-20, flash C₁₈, and reversed-phase HPLC. Their structures were identified as 3-[(3'-amino-3'-oxoprop-1'-en-2'-yl)oxy]benzamide (1), m-hydroxybenzamide (2), leptosphaepin (3), 5-methyluracil (4), feruloylamide (5), p-hydroxyphenylacetoamide (6), vanillamide (7), cyclo (L-val-L-ala) (8). Among them, 1 was a new benzamide analogue, and 2 was a new natural product. In the preliminary assays, none of the compounds 1-8 exhibited obvious inhibition of HIV-1 protease activity, and toxic with the Hela, HepG2, and U2OS cells. (IC₅₀ > 10 μmol•L⁻¹).

In vitro thrombolytic potential of bioactive compounds from marine Streptomyces sp. VITJS4 / Potencial trombolítico in vitro de compostos bioativos de Streptomyces marinhos sp. VITJS4

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.

Antibacterial and Antioxidant Activities of Acetogenins from Streptomyces sp. VE2; An Endophyte in Vernonia cinerea (L.) Less.

Strain VE2 was isolated from the stem tissue of Vernonia cinerea (L.) Less. and identified as Streptomyces sp. on the basis of morphology, chemotaxonomy and 16SrDNA sequencing. The fractionation of the crude ethyl acetate (CEA) extract from VE2 cultures led to the isolation of two acetogenins; squamocin and rollidecin B; these compounds and CEA extract had potential in antibacterial and antioxidant activities. The crude extract showed the highest activity against Salmonella typhi ATCC19430 and Bacillus cereus ATCC7064, with MIC values of 32 µg/ml. Squamocin also showed the lowest MIC (32 µg/ml) and Minimum Bactericidal Concentration (MBC) (128 µg/ml) against S. Typhi and B. cereus with corresponding large diameter of the zone of inhibitions (27.5 and 28.2 mm, respectively). Rollidecin B showed the highest DPPH antioxidant activity with SC50 value of 58.92 µg/ml.

Production, purification and characterization of L-Glutaminase from Streptomyces sp. isolated from soil.

L-Glutaminase, an amidohydrolase enzyme has been a choice of interest in the treatment of lymphoblastic leukaemia. The present study reports production, purification and characterization of extracellular glutaminase enzyme from Actinomycetes. Screening was performed for twenty Actinomycetes isolates from soil; one isolate (Isolate 2) was finally selected based on the activity of glutaminase (32.5 U/ml).The isolate was identified as Streptomyces sp. Effect of physicochemical factors namely temperature, pH, NaCl concentration, and supplementary carbon & nitrogen sources on the production of L-glutaminase from the Streptomyces sp. was carried out. The enzyme production was found to be optimum with glucose as carbon source (33 U/ml), L-glutamine as nitrogen source (33.1 U/ml), at 7 pH (32.8 U/ml), temperature 30oC (32.4 U/ml) and for 0.1% NaCl concentration (32.5 U/ml). The L-glutaminase produced from Streptomyces sp. was purified by ammonium sulphate precipitation, dialysis method and ion exchange chromatography. After the purification of the enzyme by ion exchange chromatography, it has been purified 46-fold from cell-free extract and yield was 3.25%. Characterization of extracellular L-glutaminase showed that the enzyme shown optimal activity at temperature of 30°C, pH 7, at 2% NaCl and for 0.04M substrate and the Km value was calculated to be 2.8mM and Vmax was 7.57 U/ml. The molecular weight of enzyme as determined by sodium dodecyl sulphate polyacrylamide electrophoresis (SDS-PAGE) was found to be 50 kDa.

Purification and characterization of a novel thermo stable L-methioninase from Streptomyces sp. DMMMH4 and its evaluation for anticancer activity.

L-methioninase has been purified 2.55-fold from the crude extract of Streptomyces sp. DMMMH4. The purification procedure was carried out by heat treatment and gel filtration on Sephadex G-200 column chromatography. SDS-PAGE electrophoresis showed a migrating protein band molecular mass of 47 kDa. The kinetic properties determined for the purified enzyme displayed optimum activity at 70OC and thermal stability were 70OC for 30 min. The enzyme showed maximum activity at pH 6 using acetate buffer 0.05M and was relatively stable across a broad range of pH values (5.5-8 pH). The enzyme strongly inhibited by Cr+2, Fe+2, Ni+2, Cd+2, PMSF, β-mercaptoethanol and SDS while Hg+2,Cu+2 and iodoacetate completely inhibited the enzyme activity at a final concentration of 10mM. The purified enzyme exhibited a Km of 0.7, 0.15 and 0.25 mM for L-methionine, DL-ethionine and L-cystine respectively. Cytotoxicity test demonstrate that enzyme was active against liver HepG2, breast MCF-7, lung A549, prostate PC3 and colon HCT116 cancer cell lines and has negligible toxicity toward a normal melanocyte cell line HFB4.

Antimicrobial and Anticancer Activities of Ethyl Acetate Extract of Co-culture of Streptomyces sp. ANAM-5 and AIAH-10 Isolated From Mangrove Forest of Sundarbans, Bangladesh.

In this study we investigated the antimicrobial and anticancer activities of ethyl acetate extract of co-culture of Streptomyces sp. ANAM-5 and AIAH-10 isolated from soil of mangrove forest Sundarbans, Bangladesh. The antimicrobial activity of ethyl acetate extract was determined using broth-dilution method against Candida albicans, Saccharromyces cerevaceae and Aspergillus niger whereas anticancer activity was evaluated against Ehrlich Ascites Carcinoma (EAC) cells in Swiss albino mice with the dose of 50 mg/kg and 100 mg/kg body weight (i.p). The Minimum Inhibitory Concentrations (MIC) of ethyl acetate extract was found 32μg/ml against Candida albicans while 64 μg/ml against Saccharromyces cerevaceae and Aspergillus niger. The antineoplastic activity of the crude extract was increased in dose dependent manner with a significant value (p<0.01). Bacterial crude extract enhanced the mean survival time (MST) of tumor bearing mice at 71.79% and maximum cell growth inhibition was found 75.75 % with dose of 100 mg/kg body weight (i.p.). Our study revealed that ethyl acetate extract of co-culture of Streptomyces sp. ANAM-5 and AIAH-10 is an excellent source of antimicrobial and anticancer compounds which may become helpful to treat infections and cancer.

Influence of glucose and stirring in the fermentation process in order to produce anti-Candida metabolites produced by Streptomyces sp

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.

17-16,17-Dihydroxycyclooctatinyl-hexaketide ester from Streptomyces sp. SR107 / 中国天然药物

A new hexaketide acid esterified by the 17-hydroxyl group of 16,17-dihydroxycyclooctatin, namely 17-[16,17-dihydroxycyclooctatinyl]-hexaketide ester (1), a member of the group of rare bacterial diterpenes with a fused 5-8-5 ring system was isolated from strain Streptomyces sp. SR107. The structure was determined on the basis of its spectral data (H NMR, C NMR, H-H COSY, HSQC, HMBC, NOESY, IR and HR-ESI-MS). The antibacterial activity was also evaluated in this paper.

2, 4-Di-tert-butylphenol, the bioactive compound produced by Streptomyces sp. KB1.

Streptomyces sp. KB1(TISTR 2304), identified by 16S rDNA gene, was collected from the air sample at Ao-nang, Krabi province, Thailand and deposited in the GeneBank database (accession number KF939581.1). It could produce bioactive compounds and excrete into liquid culture medium within 4 days of incubation period at 30 °C, 200 rounds per minute in shaking incubator. Bioactive compounds in culture broth supernatant were extracted with 3-times of ethyl acetate (EA), evaporated by rotary evaporator at 45 °C under reduced pressure, purified by silica gel column chromatography and eluted by gradient solvent system of ethyl acetate: hexane at ratio of 1:9, 3:7, 4:6, 6:4 and 10:0, v/v. Eight pooled fractions were obtained, name PF1 - PF8, from basis of their TLC profile. Only PF4 displayed as purified active compound which showed anti-methicillin resistant Staphylococcus aureus (MRSA) activity. The PF4 was predicted as 2, 4-Di-tert-butylphenol (C14H22O) with molecular weight 206.2.

Antifungal Substances from Streptomyces sp. A3265 Antagonistic to Plant Pathogenic Fungi

In a previous study, we identified a Streptomyces sp., A3265, as exhibiting potent antifungal activity against various plant pathogenic fungi, including Botrytis cinerea, Colletotrichum gloeosporioides, and Rhizoctonia solani. This strain also exhibited a biocontrolling effect against ginseng root rot and damping-off disease, common diseases of ginseng and other crops. In this study, we isolated two antifungal substances responsible for this biocontrolling effect via Diaion HP-20 and Sephadex LH-20 column chromatography, medium pressure liquid chromatography, and high-performance liquid chromatography. These compounds were identified as guanidylfungin A and methyl guanidylfungin A by spectroscopic methods. These compounds exhibited potent antimicrobial activity against various plant pathogenic fungi as well as against bacteria.

Antagonistic Effect of Streptomyces sp. BS062 against Botrytis Diseases

The use of microorganisms and their secreted molecules to prevent plant diseases is considered an attractive alternative and way to supplement synthetic fungicides for the management of plant diseases. Strain BS062 was selected based on its ability to inhibit the mycelial growth of Botrytis cinerea, a major causal fungus of postharvest root rot of ginseng and strawberry gray mold disease. Strain BS062 was found to be closely related to Streptomyces hygroscopicus (99% similarity) on the basis of 16S ribosomal DNA sequence analysis. Postharvest root rot of ginseng and strawberry gray mold disease caused by B. cinerea were controlled up to 73.9% and 58%, respectively, upon treatment with culture broth of Streptomyces sp. BS062. These results suggest that strain BS062 may be a potential agent for controlling ginseng postharvest root rot and strawberry gray mold disease.

Antitumor compounds from Streptomyces sp. KML-2, isolated from Khewra salt mines, Pakistan

BACKGROUND: Actinomycetes are gram positive bacteria with high G + C content in their DNA and are capable of producing variety of secondary metabolites. Many of these metabolites possess different biological activities and have the potential to be developed as therapeutic agents. The aim of the present study was to screen actinomycetes inhabiting halophilic environment such as Khewra salt mines present in Pakistan for cytotoxic and antitumor compounds. RESULTS: An actiomycetes strain designated as Streptomyces sp. KML-2 was isolated from a saline soil of Khewra salt mines, Pakistan. The strain Streptomyces sp. KML-2 showed 84 % cytotoxic activity against larvae of Artemiasalina. In the screening phase, the strain exhibited significant antitumor activity with IC50 values of 12, 48 and 56 µg/ml against Hela, MDBK and Vero cell lines, respectively. After that extract from 20 l fermentation was used to purify secondary metabolites by several chromatographic techniques. Structure elucidation of isolated compounds revealed that it is highly stable producer of Chromomycin SA (1) and 1-(1H-indol-3-yl)-propane-1,2,3-triol (2). Both of the isolated compounds showed significant antitumor activity against Hela and MCF-7 cancer cell lines (IC50 values 8.9 and 7.8 µg/ml against Hela; 12.6 and 0.97 µg/ml against MCF-7, respectively). The 16S rRNA gene sequence (1437 bp) of the strain confirm its identity (99 %) with Streptomyces griseus. CONCLUSIONS: From this research work we were successful in isolating two potent antitumor compounds, Chromomycin SA and 1-(1H-indol-3-yl)-propane-1,2,3-triol from Streptomyces KML-2 strain, isolated from Khewra salt mine. As such this is the second report which confirms that S. griseus can produce Chromomycin SA without introducing any mutagenesis in its biosynthesizing gene cluster and isolated indole derivative is being reported first time from any member of actinomycetes group with having novel antitumor activity against Hela and MCF-7 cells Nucleotide sequences: Nucleotide sequence data reported are available in the GenBank database under the accession number: GenBank KJ009562.

Antimicrobial and antiproliferative prospective of kosinostatin-a secondary metabolite isolated from Streptomyces sp / 药物分析学报

Cancer is a communal health hazard worldwide. The present investigation attempts to evaluate anti-microbial and anticancer potential of kosinostatin on mammary carcinoma cell line (MCF-7). The an-ticancer and antiproliferative activities of kosinostatin were analyzed on MCF cell line by MTT assay and cytotoxicity assays like lactate dehydrogenase (LDH) and glutathione (GSH). The secondary metabolite kosinostatin exhibited its apoptotic nature by expressing p53 protein. Collectively, the results acquired from this study promise that kosinostatin shows the potent anticancer activity.

Anti-Inflammatory Effect of Violapyrones B and C from a Marine-derived Streptomyces sp

Recently, we reported violapyrones B, C, H and I, unusual 3, 4, 6-trisubstituted alpha-pyrone derivatives, from the culture broth of the marine Streptomyces sp. 112CH148. In previous studies, violapyrones have been shown to have antibacterial and antitumor activities. However, the anti-inflammatory effect of violapyrones has not been reported yet. As part of our ongoing study for the discovery of bioactive metabolites from marine microorganisms, we found that violapyrones also have anti-inflammatory activity. In this study, we investigated the effect of violapyrones on LPS-induced inflammatory responses in vitro. Violapyrones B and C did not affect the viability of RAW 264.7 cells at concentrations up to 25 microM. However, violapyrones B and C inhibited the production of NO compared to the LPS-induced control. In addition, violapyrones B and C down-regulated the expression of iNOS protein in LPS-stimulated RAW 264.7 cells. To the best of our knowledge, this is the first report on the anti-inflammatory activity of violapyrones B and C.

Violapyrone J, alpha-Pyrone Derivative from a Marine-derived Actinomycetes, Streptomyces sp

A new alpha-pyrone derivative, violapyrone J (1), and along with the two known violapyrones B (2) and C (3) were isolated from the fermentation broth of a marine actinomycete Streptomyces sp. SC0718. The structure of violapyrone J (1) was elucidated from 1D and 2D NMR spectroscopic analyses.