Endophytic Actinomycetes Research Articles

Diversity and Antimicrobial Activity of Endophytic Actinomycetes Isolated from Plant Roots in Thailand

In this study, 37 endophytic actinomycetes were isolated from roots of 14 plant species obtained in Thailand, where biological diversity is known to be high. The isolates were identified as members of the genera Micromonospora (21 isolates), Streptomyces (14 isolates), Plantactinospora (1 isolate), and Polymorphospora (1 isolate) based on their phenotypic characteristics and the 16S rRNA gene sequence . The most frequent isolates (56.8%) were members of the genus Micromonospora. The antimicrobial activity screening showed that seven Micromonospora isolates and eight Streptomyces isolates were active against Bacillus subtilis ATCC 6633, Kocuria rhizophila ATCC 9341 and Staphylococcus aureus ATCC 6538p. To our knowledge, the isolates showed the difference of antimicrobial activity based on the production media. Moreover, this study suggested Thai plant species to be valuable sources of interesting endophytic actinomycetes producing antimicrobial metabolites.

Pseudonocardia broussonetiae sp. nov., an endophytic actinomycete isolated from the roots of Broussonetia papyrifera.

A novel endophytic actinomycete, designated strain Gen 01T, was isolated from the roots of Broussonetia papyrifera and characterized by using a polyphasic approach. The predominant cellular fatty acids were iso-C16 : 0, summed feature 3, iso H-C16 : 1, C16 : 0 and iso-C14 : 0. The polar lipid profile consisted of diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol mannosides, phospholipids of unknown structure containing glucosamine inositol, phosphatidylinositol and unidentified phospholipids. The major menaquinone was MK-8 (H4). The DNA G+C content of the genome sequence, consisting of 7 177 725 bp, was 74.5 mol%. Phylogenetic analysis of the full-length 16S rRNA gene sequences showed that strain Gen 01T belongs to the genus Pseudonocardia with the highest sequence similarity to Pseudonocardia petroleophila CGMCC 4.1532T (98.9 %) and lower than 98.7 % similarity to other species of the genus Pseudonocardia with validly published names. The average nucleotide identity and digital DNA-DNAhybridization values between strain Gen 01T and P. petroleophila CGMCC 4.1532T were 84.6 and 30.9 %, respectively. Furthermore, the morphological, physiological and biochemical characteristics were sufficient to categorize strain Gen 01T as being distinct from P. petroleophila CGMCC 4.1532T. Consequently, based on phenotypic and genotypic characteristics, strain Gen 01T represents a novel species of the genus Pseudonocardia, for which the name Pseudonocardia broussonetiae sp. nov. is proposed. The type strain is Gen 01T (=CICC 24820T=JCM 33840T).

Characterisation of Two Polyketides from Streptomyces sp. SKH1-2 Isolated from Roots of Musa (ABB) cv. 'Kluai Sao Kratuep Ho'.

An endophytic actinomycete strain SKH1-2 isolated from Musa (ABB) cv. 'Kluai Sao Kratuep Ho' collected in Suphan Buri province (14° 54' 22.5″ N/100° 04' 50″ E), Thailand, was identified as Streptomyces pseudovenezuelae based on phenotypic and chemotaxonomic characteristics, and 16S rRNA sequence analyses. A chemical investigation led to the isolation of two polyketide molecules from the n-butanol crude extract of the strain SKH1-2 culture broth. The compounds were purified using various chromatographic techniques and identified using spectroscopic methods compared with earlier published data. Compound 1, chartreusin, is known as an anti-Gram (+) bacterial compound and was active against Bacillus subtilis ATCC 6633, Kocuria rhizophila ATCC 9341 and Staphylococcus aureus ATCC 6538p with MIC values of 3.1, 1.6 and 12.5μg/mL, respectively. Compound 2, lumichrome, did not show activity against all tested microbes. To our knowledge, this is the first report of chartreusin and lumichrome isolated from S. pseudovenezuelae. Taken together, it could be proved that Thai plant species are valuable reservoirs of interesting endophytic actinomycetes producing several interesting biologically active compounds.

Gordonia oryzae sp. nov., isolated from rice plant stems (Oryza sativa L.).

A novel endophytic actinomycete, designated strain RS15-1ST, was isolated from surface-sterilized stems of Oryza sativa L. collected from Sisaket province, Thailand. The colony of strain was strong orange, catalase-positive and oxidase-negative. Growth occurred at a temperature range of 17-37 °C, at pH 4.0-9.0 and in the presence of 0-13 % (w/v) NaCl. Phylogenetic analyses based on the 16S rRNA sequences showed that strain RS15-1ST belonged to the genus Gordonia and was closely related to Gordonia polyisoprenivorans DSM 44302T (98.8 %) and Gordonia rhizosphera DSM 44383T (98.4 %). The major cellular fatty acids were C16 : 0, C18 : 0 10-methyl (tbsa), C16 : 1ω7c/C16 : 1ω6c and C18 : 1ω9c. The menaquinones were MK-9(H2) and MK-8(H2). Strain RS15-1ST contained meso-diaminopimelic acid, arabinose, galactose, mannose and ribose in whole-cell hydrolysates. The polar lipids of the strain were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol mannosides, an unidentified polar lipid and two unidentified phospholipids. The DNA G+C content was 66.3 mol%. In silico DNA-DNA hybridization of strain RS15-1ST showed 48.3 and 20.5 % relatedness to its closest neighbours, Gordonia polyisoprenivorans DSM 44302T and Gordonia rhizosphera DSM 44383T, respectively. Based on data of genotypic, phenotypic, phylogenetic and chemotaxonomic analysis, strain RS15-1ST represents a novel species of the genus Gordonia, for which the name Gordonia oryzae sp. nov. is proposed. The type strain is RS15-1ST (=TBRC 8485T=NBRC 113446T).

Endophytic actinomycetes Streptomyces spp mediated biosynthesis of copper oxide nanoparticles as a promising tool for biotechnological applications.

In this study, two endophytic actinomycetes isolates Oc-5 and Acv-11, were isolated from healthy leaves of medicinal plant Oxalis corniculata L. These isolates were identified as Streptomyces zaomyceticus Oc-5 and Streptomyces pseudogriseolus Acv-11 using 16S rRNA gene sequence. Biomass extract of these strains were used as a greener attempt for synthesis of copper oxide nanoparticles (CuO-NPs). The synthesized NPs were characterized by UV-Vis spectroscopy, Fourier transform infra-red (FT-IR) spectroscopy, X-ray diffraction (XRD)' transmission electron microscopy (TEM), energy dispersive X-ray (EDX) and X-ray photoelectron spectroscopy (XPS). Green synthesized NPs showed surface plasmon resonance (SPR) absorption band at 400nm, crystalline nature, spherical-shaped with an average size of 78nm and 80.0nm for CuO-NPs synthesized using strain Oc-5 and Acv-11, respectively. The bioactivities of CuO-NPs were evaluated. Results revealed that CuO-NPs exhibited promising antimicrobial activity against prokaryotic and eukaryotic microbial cells (Gram positive bacteria, Gram negative bacteria, unicellular and multicellular fungi). In addition, it showed antimicrobial potential against phyto-pathogenic fungal strains Fusarium oxysporum, Pythium ultimum, Aspergillus niger and Alternaria alternata. We further explored the in vitro antioxidant activity and cytotoxicity for biosynthesized CuO-NPs. The results revealed that' scavenging and total antioxidant activity for NPs synthesized using Streptomyces pseudogriseolus Acv-11 was better than those synthesized by Streptomyces zaomyceticus Oc-5. Also, the morphological changes and cell viability for Vero and Caco-2 cell line due to NPs treatments were assessed using MTT assay method. Furthermore, Larvicidal efficacy against Musca domestica and Culex pipiens was evaluated. The results obtained in this study clearly showed that biosynthesized CuO-NPs exhibited effective bioactivity and, therefore, provide a base for the development of versatile biotechnological applications soon.

Applying Experimental Design for Low-Cost and Eco-Friendly Biosynthesis of MgONPs from Local Endophytic Actinomycetes as an Antiphytopathogens Agent

Applying Experimental Design for Low-Cost and Eco-Friendly Biosynthesis of MgONPs from Local Endophytic Actinomycetes as an Antiphytopathogens Agent

Effects of Biofertilizer Produced from Bradyrhizobium and Streptomyces griseoflavus on Plant Growth, Nodulation, Nitrogen Fixation, Nutrient Uptake, and Seed Yield of Mung Bean, Cowpea, and Soybean

The use of biofertilizers is important for sustainable agriculture, and the use of nodule bacteria and endophytic actinomycetes is an attractive way to enhance plant growth and yield. This study tested the effects of a biofertilizer produced from Bradyrhizobium strains and Streptomyces griseoflavus on leguminous, cereal, and vegetable crops. Nitrogen fixation was measured using the acetylene reduction assay. Under N-limited or N-supplemented conditions, the biofertilizer significantly promoted the shoot and root growth of mung bean, cowpea, and soybean compared with the control. Therefore, the biofertilizer used in this study was effective in mung bean, cowpea, and soybean regardless of N application. In this study, significant increments in plant growth, nodulation, nitrogen fixation, nitrogen, phosphorus, and potassium (NPK) uptake, and seed yield were found in mung beans and soybeans. Therefore, Bradyrhizobium japonicum SAY3-7 plus Bradyrhizobium elkanii BLY3-8 and Streptomyces griseoflavus are effective bacteria that can be used together as biofertilizer for the production of economically important leguminous crops, especially soybean and mung bean. The biofertilizer produced from Bradyrhizobium and S. griseoflavus P4 will be useful for both soybean and mung bean production.

Extraction and partial purification of secondary metabolites from endophytic actinomycetes of marine green algae Caulerpa racemosa against multi drug resistant uropathogens

Abstract The aim of this study is to identify potential antibacterial compounds of endophytic actinomycetes (EA) from marine algae Caulerpa racemosa for inhibit the multi drug resistant (MDR) Gram negative bacteria (GNB) including Eschericia coli (E. coli), Pseudomonas aeruginosa (P. aeruginosa), Proteus mirabilis (P. mirabilis), Klebsiella pnumonia (K. pnumoniae) and Enterobacter sp., which were confirmed by UTI panel antibiotic discs. Among the 100 EA, five strains were showed better antogonistic activity against all the selected uropathogens. In the ethyl acetate extract, the NMS 5 strains of EA showed excellent anti-bacterial activity against all the MDR uropathogen. Further, the NMS 5 extract was partially purified from thin layar chramatogaraphy (TLC) and spot 3 of TLC crude compound showed 14, 12 mm zone of inhibition against both the uropathogens. In addition, the minimum inhibition concentration (MIC) of crude exhibited the inhibition percentage was 87 and 84 at 120 µg/mL concentration. The MIC was confirmed by minimum bactericidal concentration (MBC) and it was indicated same as MIC. The treated NMS 5 crude compound of P. aeruginosa and K. pneumonia cells were showed more dead cells, intracellular damage and morphological damage with disaggregated cells by confocal laser scanning electron microscope (CLSM) and scanning electron microscope (SEM). Hence, the present study was revealed that the algae derived EA have the potential anti-bacterial metabolites for inhibition of MDR bacteria and it can be applied for various other infections after further purification.

Endophytic Actinomycetes as Potential Producers of Hemicellulases and Related Enzymes for Plant Biomass Degradation

Abstract Tailor made enzymatic preparation must be design to hydrolyze efficiently plant biomass, once that each plant biomass possesses a distinct cell wall composition. Most of actinomycetes used for plant cell wall degradation are focused on the cellulases and xylanases production. However, a wide range of enzymes must be produced for an efficient degradation of lignocellulose materials. During the last decade several unusual environments were studied to obtain strains that produce glycohydrolases with innovator characteristics. In this context, the present work concerned the selection of endophytic actinomycetes as producers of hemicellulases and related enzymes with different enzymatic profiles, for use in the deconstruction of lignocellulosic biomass. A total of 45 Brazilian actinomycetes previously isolated from plants (endophytics) and soil were prospected for hemicellulases and β-glucosidase production. Four strains highlighted for hemicellulase production (DR61, DR63, DR69 and DR66) and were selected for cultivation under other inductors substrates (xylan and pectin). All strains belong to Streptomyces genera and have their extracts tested for degradation of several hemicellulolytic substrates. The strains presented different glicohydrolyse enzymes profiles mainly for xylans and glucans that can be used for specific formulations of enzymes applied on the biomass deconstruction, principally on sugar cane bagasse.

Determining the Anti-malarial potential of an unknown Endophytic Streptomyces species BJSG4 isolated from Kalanchoe pinnata (Lam.) Pers.

Malaria is the horrible disease caused by Plasmodium species. Present anti-malarial drug resistant issues alarming the felt need of new drug molecules and research to control the causative agents. So, the present study investigates the antimalarial potential of the Streptomyces sp. BJSG4 isolated from the ethnobotanical medicinal plant Kalanchoe pinnata (Lam.)Pers. Stem of Kalanchoepinnata (Lam.) Pers. was used as an isolation source. The isolates were confirmed by traditional and conventional methods. Anti-malarial screening was done against Plasmodium falciparum 3D7 strain under in-vitro condition and the activity was analyzed in GraphPad Prism software (demo version 6. 0). A new endophytic Streptomyces species named BJSG4 was isolated from Kalanchoe pinnata (Lam.) Pers., an ethnobotanical medicinal plant and a host of the endophyte. The presence of the compound 3-hydroxy-1-(4-{13-(4-(3-hydroxy-3-phenylacryloyl) phenyl) tridecyl}-phenyl)-3-phenylprop-2-en-1-one in the endophyte BJSG4was confirmed by gas chromatography-mass spectrometry. The purified compound exhibited in-vitro activity against the malaria parasite Plasmodiumfalciparum3D7 (log IC50 value: 3. 47 nM). A statistical correlation coefficient (R2) value of 0. 9 confirmed the highest level of positive in vitro interaction between the compound and the malaria parasite. Culture-dependent and -independent methods confirmed that the active compound-producing endophyte BJSG4 belongs to the genus Streptomyces. The present study proved that ethnobotanical medicinal plants can be the source of potentially novel drugs produced by endophytic actinomycetes.

Secondary Metabolites of Endophytic Actinomycetes: Isolation, Synthesis, Biosynthesis, and Biological Activities.

Endophytic Actinobacteria are a microbial group that is still poorly investigated. Their association with plants constitutes a unique trait conferring specific biological and chemical features to endophytic Actinobacteria. This contribution discusses aspects of endophytic actinobacterial biology and chemistry comprehensively, including the biosynthesis and total synthesis of secondary metabolites produced in culture. It also presents perspectives for the future of microbial bioactive natural products discovery, with emphasis on the secondary metabolism of endophytic Actinobacteria.

Secondary metabolites produced by Microbacterium sp. LGMB471 with antifungal activity against the phytopathogen Phyllosticta citricarpa.

The citrus black spot (CBS), caused by Phyllosticta citricarpa, is one of the most important citrus diseases in subtropical regions of Africa, Asia, Oceania, and the Americas, and fruits with CBS lesions are still subject to quarantine regulations in the European Union. Despite the high application of fungicides, the disease remains present in the citrus crops of Central and South America. In order to find alternatives to help control CBS and reduce the use of fungicides, we explored the antifungal potential of endophytic actinomycetes isolated from the Brazilian medicinal plant Vochysia divergens found in the Pantanal biome. Two different culture media and temperatures were selected to identify the most efficient conditions for the production of active secondary metabolites. The metabolites produced by strain Microbacterium sp. LGMB471 cultured in SG medium at 36°C considerably inhibited the development of P. citricarpa. Three isoflavones and five diketopiperazines were identified, and the compounds 7-O-β-D-glucosyl-genistein and 7-O-β-D-glucosyl-daidzein showed high activity against P. citricarpa, with the MIC of 33μg/mL and inhibited the production of asexual spores of P. citricarpa on leaves and citrus fruits. Compounds that inhibit conidia formation may be a promising alternative to reduce the use of fungicides in the control of CBS lesions, especially in regions where sexual reproduction does not occur, as in the USA. Our data suggest the use of Microbacterium sp. LGMB471 or its metabolites as an ecological alternative to be used in association with the fungicides for the control of CBS disease.

Classification and characterization of endophytic actinomycetes isolated from cinnamomum cassia presl

Currently, antibiotic resistance in pathogenic bacteria is a significant clinical problem with the increase of deseases and a serious public health concern. Thus, the identification of new antimicrobial agents, especially the secondary metabolites products by endophytic actinobacteria from medical plants could be promising sources of biologically active compounds in medical fields. This study focused on identification and evaluation of antimicrobial activity against pathogens; genes involved in their secondary metabolisms, and screening of anthracycline producing capacity (mainly presented in anti-cancer antibiotics) of YBQ75 isolated from Cinnamomum cassia Presl. plants in Yen Bai province. Based on manual of bacterial classification, method in International Streptomyces Project (ISP) and the 16S rRNA gene sequence (GenBank Acc. No. KR814822), the endophytic actinomycetes YBQ75 was named Streptomyces cavourensis YBQ75 with 100% identity. The strain S. cavourensis YBQ75 showed the remarkable antibacterial activities against 5 tested pathogens (Salmonella enterica ATCC 14028 (22.0 mm); Pseudomonas aeruginosa CNLM (19.3 mm); Staphylococcus epidermidis ATCC 12228 (19.3 mm); Enterobacter aerogenes ATCC 13048 (17.7 mm) and Proteus vulgaris CNLM (16.3 mm)) in the total of 9 tested pathogens. The detection of genes involved in antibiotic synthesis indicated that the strain S. cavourensis YBQ75 consists of all three genes related to antibiotic synthesis including polyketide synthase (pks-I) type I, polyketide synthase type II (pks-II) and nonribosomal peptide synthetase (nrps). Premarilly result showed that the strain S. cavourensis YBQ75 also present as an anthracycline productive actinomycetes. The resutls demonstrated that the endophytic actinomycetes S. cavourensis YBQ75 from medical plants could be promising sources for the production of antibiotics and anthracycline anticancer compounds.

Quadrisphaera oryzae sp. nov., an endophytic actinomycete isolated from leaves of rice plant (Oryza sativa L.).

A novel endophytic actinobacterium, designated strain RL12-1ST, was isolated from surface-sterilized leaves of Oryza sativa L. collected from Roi Et province, Thailand. Taxonomic position of strain RL12-1ST was studied using the polyphasic approach. Phylogenetic analyses base on the 16S rRNA gene sequences showed that strain RL12-1ST belonged to the genus Quadrisphaera and was closely related to Quadrisphaera granulorum AG019T (99.0%). The colony of strain RL12-1ST was vivid orange. Growth occurred at a temperature between 17-37 °C and pH between 6.0-8.0. The strain contained meso-diaminopimelic acid and galactose, glucose, mannose, ribose and rhamnose in whole-cell hydrolysates. The predominant menaquinone was MK-8(H2). The major fatty acids were C16:1ω7c/C16:1ω6c, C16:0 and C18:1ω7c. The polar lipids of the strain were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol and phosphatidylinositol mannosides. The DNA G + C content of genomic DNA was 76.1 mol%. Based on comparative analysis of physiological, biochemical and chemotaxonomic data, including DNA-DNA hybridization, strain RL12-1ST represents a novel species of the genus Quadrisphaera, for which the name Quadrisphaera oryzae sp. nov. is proposed. The type strain is RL12-1ST ( = TBRC 8486T = NBRC 113447T).

Research on rhizospheric and endophytic actinomycetes in medicinal tree peony(Paeonia suffruticosa) from five producing regions

In this study, Paeonia suffruticosa roots and rhizospheric soil in five geographic regions which were harvested in October were utilized as experimental materials, then the diversity of endophytic and rhizospheric actinomycetes were investigated by High-throughput sequencing technique. The 1 754 operational taxonomic units (OTUs) were obtained from 129 954 high quality sequences, 1 311 OTUs were detected in rhizospheric actinomycetes and belonged to four classes, four orders, twenty-seven families and ninety-seven genera, thirty-three genera such as Ilumatobacter were found in the five regions rhizospheric soil while three genera such as Longispora were only detected in the Dao-di regions, the dominant genera were Mycobacterium, Nocardioides, Streptomyces. 443 OTUs were obtained in roots and distributed in three classes, three orders, twenty-four families and fifty genera, thirteen genera such as Cryptosporangium were found in the five regions roots while Planosporangium, Luteococcus were only detected in the Dao-di regions, the dominant genera were Nocardioides. Alpha diversity analysis showed that the Shannon and Chao1 index in rhizospheric actinomycetes in Bozhou, Tongling and Nanling region were higher than Heze and Luoyang. Based on principal co-ordinates analysis (PCoA) analysis, the rhizospheric actinomycetes formations were similar in Tongling and Nanling region, at the same in Tongling and Luoyang endophytic actinomycetes. According to heatmap analysis, Bozhou, Tongling and Nanling region rhizospheric actinomycetes showed a close similarity in actinomycetes community structures on phylogenetic analysis, while Tongling, Luoyang and Nanling endophytic actinomycetes showed the same. Our results not only suggested that the rich and diverse actinomycetes resources in P. suffruticosa roots and rhizospheric soil but also revealed rhizospheric actinomycetes in the Dao-di regions had high similarity.

Endophytic Actinomycetes from Tea Plants (Camellia sinensis): Isolation, Abundance, Antimicrobial, and Plant-Growth-Promoting Activities.

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.

Molecular identification and structural characterization of marine endophytic actinomycetes Nocardiopsis sp. GRG 2 (KT 235641) and its antibacterial efficacy against isolated ESBL producing bacteria

The present study was designed to identify the potential bioactive compound from endophytic actinomycetes (EA) Nocardiopsis sp. GRG 2 (KT 235641) against selected extended spectrum beta lactamase (ESBL) producing Pseudomonas aeruginosa (P. aeruginosa) and Klebsiella pneumoniae (K. pneumoniae). Initially, the multi drug resistance (MDR) effect of selected uropathogens was confirmed by respective UTI panel of Hexa antibiotics disc methods. The zone of inhibition ≤22 mm for ceftazidime, ≤ 27 mm for cefotaxime and ≤8 mm zone of MIC stripe against both the uropathogens of phenotypic methods confirmed, the selected strains were ESBL producer. Among the various EA extracts, GRG 2 extract showed excellent antibacterial activity against both ESBL producing P. aeruginosa and K. pneumonia by agar well diffution method. The molecular identification of selected GRG 2 strain was named as Nocardiopsis sp. GRG 2 (KT235641). The antibacterial metabolites present in the TLC elution was exhibited at 274 nm by UV visible spectrometer. The partial purification of preparative HPLC fraction 3 showed 14, 16 mm against P. aeruginosa and K. pneumoniae, respectively. Based on the antibacterial effect, the FT-IR, GC-MS and LC-MS analysis of fraction 3 was confirmed as 1, 4-diaza-2, 5-dioxo-3-isobutyl bicyclo[4.3.0]nonane (DDIBN). Further, the dose dependent inhibition of DDIBN against both ESBL producing pathogens was observed at 75 μg/mL by minimum inhibition concentration (MIC) and minimum bactericidal concentration (MBC). The increased cell death and disrupted cell membrane integrity were observed at MIC of DDIBN by confocal laser scanning electron microscope (CLSM) and scanning electron microscope (SEM). The results were proved that the DDIBN has potential antibacterial metabolites against ESBL producing pathogens and it can be applied for various other biomedical fields.

Antibacterial and anticancer potential of marine endophytic actinomycetes Streptomyces coeruleorubidus GRG 4 (KY457708) compound against colistin resistant uropathogens and A549 lung cancer cells

The aim of the current study is to identify bioactive compound from marine endophytic actinomycetes (MEA) isolated from Gulf of Mannar region, Southeast coast of India. Among the isolated actinomycetes, strain GRG 4 exhibited excellent ability to inhibit isolated colistin resistant (CR) Pseudomonas aeruginosa (P. aeruginosa) and Klebsiella pneumoniae (K. pneumoniae), which is a emerging threat to the world. The strain was identified as Streptomyces coeruleorubidus GRG 4 (KY457708), based on morphological, biochemical, phenotypic and genotypic characters. The bioactive metabolites present in the methanolic extract were partially purified by TLC and preparative HPLC. The active HPLC fraction 2 showed 15, 20 mm zone of inhibition against both CR P. aeruginosa and K. pneumoniae respectively. Analytical HPLC and FT-IR results of fraction 2 showed with carbonyl group. Both GC-MS and LC-MS results confirmed that the fraction 2 contained chemical constituents of Bis (2-Ethylhexyl) Phthalate (BEP). The compromised structure with loosely integrated and ruptured cell wall of BEP treated CR bacteria were observed by confocal laser scanning microscope (CLSM) and scanning electron microscope (SEM) at 75 μg/mL of minimum inhibitory concentration (MIC) dose. Further, cytotoxic effect of BEP against A549 human lung cancer cells revealed complete inhibition by cell proliferation and apoptosis was observed at 100 μg/mL in 24 h treatment. In addition, irreversible ROS dependent oxidative damage was clearly observed at the IC50 concentration of BEP. The toxicity of BEP was also studied against Vibrio fischeri (V. fischeri) and found to be highly toxic after 15 and 30 min of treatment. Based on the results it could be concluded that the identified compound BEP is a potent inhibitor for CR bacteria and A549 lung cancer cells.

Transcriptome analysis of Valsa mali reveals its response mechanism to the biocontrol actinomycete Saccharothrix yanglingensis Hhs.015

BackgroundApple canker is a devastating branch disease caused by Valsa mali (Vm). The endophytic actinomycete Saccharothrix yanglingensis Hhs.015 (Sy Hhs.015) can effectively inhibit the growth of Vm. To reveal the mechanism, by which Vm respond to Sy Hhs.015, the transcriptome of Vm was analyzed using RNA-seq technology.ResultsCompared with normal growing Vm in the control group, 1476 genes were significantly differentially expressed in the Sy Hhs.015’s treatment group, of which 851 genes were up-regulated and 625 genes were down-regulated. Combined gene function and pathway analysis of differentially expressed genes (DEGs) revealed that Sy Hhs.015 affected the carbohydrate metabolic pathway, which is utilized by Vm for energy production. Approximately 82% of the glycoside hydrolase genes were down-regulated, including three pectinase genes (PGs), which are key pathogenic factors. The cell wall structure of Vm was disrupted by Sy Hhs.015 and cell wall-related genes were found to be down-regulated. Of the peroxisome associated genes, those encoding catalase (CAT) and superoxide dismutase (SOD) which scavenge reactive oxygen species (ROS), as well as those encoding AMACR and ACAA1 which are related to the β-oxidation of fatty acids, were down-regulated. MS and ICL, key genes in glyoxylate cycle, were also down-regulated. In response to the stress of Sy Hhs.015 exposure, Vm increased amino acid metabolism to synthesize the required nitrogenous compounds, while alpha-keto acids, which involved in the TCA cycle, could be used to produce energy by deamination or transamination. Retinol dehydrogenase, associated with cell wall dextran synthesis, and sterol 24-C-methyltransferase, related to cell membrane ergosterol synthesis, were up-regulated. The genes encoding glutathione S-transferase, (GST), which has antioxidant activity and ABC transporters which have an efflux function, were also up-regulated.ConclusionThese results show that the response of Vm to Sy Hhs.015 exposure is a complicated and highly regulated process, and provide a theoretical basis for both clarifying the biocontrol mechanism of Sy Hhs.015 and the response of Vm to stress.

Actinomycin-Producing Endophytic Streptomyces parvulus Associated with Root of Aloe vera and Optimization of Conditions for Antibiotic Production.

Endophytic actinomycetes are a rich source of novel antimicrobial compounds. The aim of this study was to evaluate the production of antimicrobial compound by endophytic Streptomyces sp. Av-R5 associated with root of Aloe vera against multidrug-resistant human pathogens. The 16S rRNA sequence of the isolate Av-R5 has been identified as Streptomyces parvulus NBRC 13193T (AB184326) and the sequence was submitted to the National Center for Biotechnology Information (NCBI) GenBank database (accession number KY771080). Streptomyces parvulus Av-R5 grown under submerged fermentation condition optimized by central composite design (glucose 11.16g/L, soybean meal 10.25g/L, sodium chloride 11.18g/L, calcium carbonate 1.32g/L at pH7.19 at 31.42°C with 6.04% seed inoculum for 10days of incubation) exhibited the highest activity against multidrug-resistant Staphylococcus aureus JNMC-3, Staphylococcus epidermidis JNMC-4, Klebsiella pneumoniae MTCC-3384, Klebsiella pneumoniae JNMC-6, Pseudomonas aeruginosa MTCC-741, Proteus vulgaris JNMC-7, Candida albicans MTCC-183, and Aspergillus niger MTCC-872. The structures of the active compounds were elucidated by UV-Vis spectroscopy, 1H and 13C NMR, FT-IR, and ESIMS. Actinomycin D and actinomycin X0β were detected in crude extracts and major components were eluted by HPLC at 10.96 and 6.81min, respectively. In this case, a high yield of actinomycin D and actinomycin X0β (400mg/L) was achieved with Streptomyces parvulus Av-R5, fermented in glucose soybean meal broth media, which can be used in industrial fermentation process to obtain high yields.