Actinobacteria Basics and Biotechnological Applications

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The screening was focused on the actinomycetes that showed the ability to produce antibacterial, antifungal, and extracellular enzymes such as amylase, cellulase, and protease. The antagonistic assay was tested against Escherichia coli, Staphylococcus aureus, Sclerotium rolfsii and Fusarium oxysporum. Isolate JKP-8 was an isolate that showed the highest activity in inhibiting the growth of E.

Isolate JKP-5 showed the highest activity in inhibiting the growth of F. There were no actinomycetes isolates that showed an ability to inhibit the growth of S. Antifungal production of a strain of Actinomycetes spp. Indonesian Journal of Biotechnology 16 1 :1— Actinomycetes from Capsicum annuum L. Characterization of rhizospheric soil streptomycetes from Moroccan habitats and their antimicrobial activities. World Journal of Microbiology and Biotechnology — Amplification of DNA from native populations of soil bacteria by using the polymerase chain reaction.

Applied and Environmental Microbiology — Chemical composition of hyphal and sclerotial walls of Sclerotium rolfsii Sacc. Canadian Journal of Microbiology — Antagonistic bacteria against Schizophyllum commune Fr. Biotropia — Evaluation of actinomycete isolates obtained from herbal vermicompost for the biological control of Fusarium wilt of chickpea.

Crop Protection — Growth promotion and protection against damping-off of wheat by two rock phosphate solubilizing actinomycetes in a P-deficient soil under greenhouse conditions. Applied Soil Ecology — Hayakawa M, Nonomura H. Humic acid-vitamin agar, a new medium for the selective isolation of soil actinomycetes. Journal of Fermentation Technology — Rhizosphere interactions: root exudates, microbes, and microbial communities. Botany — Identification of actinomycetes from plant rhizospheric soils with inhibitory activity against Colletotrichum spp.

BMC Research Notes Proteolytic actinomycetes from Indian habitats. Journal of Culture Collections — Kanti A. Biodiversitas — Isolation and screening of cellulolytic actinomycetes from diverse habitats. Seperation, identification and analysis of pigment melanin production in Streptomyces. Embrapa Solos. Rio de Janeiro. Antimicrobial properties of actinomycetes from the soil of Himalaya.

Non-streptomycete actinomycetes as biocontrol agents of soil-borne fungal plant pathogens and as plant growth promoters. Streptomyce morphogenetics: Dissecting differentiation in a filamentous bacterium. Identification and use of actinomycetes for enhanced nodulation of soybean co-inoculated with Bradyrhizobium japonicum. Different pre-treatment methods in selective isolation of Actinomycetes from mangrove sediments of Ariyankuppam back water estuary, Puducherry. Actinomycetes as antagonists of litter decomposer fungi.

Karlovsky P Secondary Metabolites in Soil Ecology.


Editoral Premier. Kishore P Isolation, characterization and identification of Actinobacteria of Mangrove ecosystem, Bhitarkanika, Odisha. Dissertation, National Institute of Technology Rourkela. Isolation and screening of soil Actinomycetes as source of antibiotics active against bacteria. Antibacterial activity of some actinomycetes from Tamil Nadu, India. A comparative study of the activity of fifty antibiotic actinomycetes against a variety of soil bacteria. Pyrosequencing-based assessment of soil pH as a predictor of soil bacterial community structure at the continental scale.

Microbial populations cultivable litter soil and of a storage unit in brazilian semi-arid. Molecular identification and characterization of antimicrobial active actinomycetes strains from some Egyptian soils. Marine actinobacterial metabolites: current status and future perspectives. Rhizobium tropici, a novel species nodulating Phaseolus vulgaris L.

Actinobacteria Basics And Biotechnological Applications

Beans and Leucaena sp. BMC Mol. Isolation of rhizospheric and roots endophytic actinomycetes from Leguminosae plant and their activities to inhibit soybean pathogen, Xanthomonas campestris pv. FEMS Microbiol. Amplicons were confirmed on 1.

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Big Dye sequencing reaction was then performed followed by a secondary clean-up prior to submission to UW Biotech for analysis University of Wisconsin-Madison. Hits with the best bitscore were assigned to that clade. Isolates were inoculated along the center of wells.

Actinomycetes (KINGDOM MONERA )

Inhibition was scored in binary, as 0 no inhibition or 1 inhibition. For a subset of isolates, experimental wells were assigned a rating from 0 to 3 depending on the level of inhibition 0—no inhibition, 1—slight inhibition, 2—presence of a zone of inhibition, 3—complete inhibition.

Inhibition fraction was determined by taking the average inhibition of Strain X vs. Pathogen Y to account for variance if multiple X by Y challenges were run. This was done for all pathogens against X. Gram-positive, gram-negative, and fungal averages were then computed to find the inhibition fraction for X. Fuzzy clustering was used to group inhibitory profiles using Mfuzz Cultures were grown in rich medium supplemented with 0. Cells were washed with Neutral phenol and chloroform were added, and tubes were gently shaken until uniformly white. Tubes were gently mixed until DNA appeared.

DNA was pelleted, supernatant was removed, and the pellet was resuspended in TE with 0. DNA was quantified, checked for purity, and run on a gel to verify high molecular weight. Genes were called with prodigal v2. HMMER v3. Alignments were then converted to codon alignments and concatenated. The multi-locus phylogeny was generated using RAxML v8. BGCs were determined to be full-length if 0. Shannon entropy with respect to phylogeny, strain source e.

Streptomyces were cultured on YPM agar media and grown for up to 14 days or until sporulation. Extracts were transferred to new vials and vacuum dried. Both instruments were operated using Bruker Hystar software. The column was set to initial conditions in 0. The flow rate was 0. Automatic internal calibration was conducted after each run by introducing Tune Mix Agilent, ESI-L low concentration through a divert valve during re-equilibration. Buckets were normalized to the sum of buckets values in the analysis. Assays were performed in well plates using sheep blood 0.

Thirty microliters of supernatant were transferred to a clear plate and OD was read. An increase in OD indicated the red blood cell lysis and hemolytic activity. All mouse experiments and protocols received ethical approval from the University of Wisconsin Institutional Animal Care and Use Committee. Animals were maintained in accordance with the criteria of the Association for Assessment and Accreditation of Laboratory Animal Care.

Middleton Memorial Veterans Hospital. Previous studies have shown neutropenia neutrophils, per mm 3 in this model for the h study period The final inoculum was adjusted to a 0. Fungal counts of the inoculum determined by viable counts of C. Disseminated infection with the Candida was achieved by injection of 0. Animals were sacrificed by CO 2 asphyxiation.

Kidneys of each mouse were removed and placed in sterile 0. Results were expressed as the mean number of CFU per kidney for three mice. No-treatment and zero-hour controls were included in all experiments. Animals for the present studies were maintained in accordance with the criteria of the Association for Assessment and Accreditation of Laboratory Animal Care. Middleton Memorial VA Hospital. Previous studies have shown that this regimen produces neutropenia in this model for 5 days After dilution into fresh Mueller—Hinton broth, the bacterial counts of the inoculum ranged from 10 7.

Thigh infections with each of the isolates were produced by injection of 0. No-treatment controls were included in all experiments. To analyze fractions as candidates for in vivo study, we used Core-shell technology UPLC columns which provide extensive sensitivity and dynamic range for small molecules 60 Fig. Additionally, NMR data for each fraction were acquired using a 1.

Prior to in vivo testing, samples were analyzed by NMR and LCMS to ensure characteristics of each sample matched to the original sample that exhibited in vitro activity. Acquisitions were performed such that ion counts of 10 6 or greater were observed for the highest abundance ions, which facilitated detection of minor components. As such, ion counts in the millions maintain accuracy and appropriate peak shape.

Some lipids e. Filtered HP20 was washed with water and extracted with acetone.

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These four fractions were further purified by HPLC to generate 80 fractions for each strain in a well plate. High-throughput screening was used to evaluate the antimicrobial activity of these fractions. Dereplication based on 1. Select strains were re-grown in the large scale in the three media, and followed the same processor mentioned before to generate the fractions containing potential novel active compounds. These fractions were formulated and tested in vivo activity Fig. MIC values of cyphomycin were defined as the lowest concentration at which a prominent decrease in growth turbidity i.

Median MIC from replicate assays is reported. The HP20 was filtered and washed with distillated water and soaked with acetone. Organic phase was separated and dried to give the crude extract 2. The 13 C NMR spectrum showed 77 signals assigned to 12 methyl, 18 methylene, 36 methine, 5 carbonyl carbons, 4 tertiary carbons sp 2 and 2 quaternary carbon groups by multiplicity-edited g HSQC experiment.

Geometry of the two double bonds of macrolide were determine to be E configured by coupling constants of 3 J H-2,H-3 Compound 2 was named cyphomycin. Genomic data can be found at DOI: A modified version of the southern pine beetle Fig. Vallery is used under the Creative Commons Attribution 3. Photos of Cyphomyrmex Figs. Sprenger, M. New mechanisms, new worries. Science , — Brown, E. Antibacterial drug discovery in the resistance era.

Nature , — Newman, D. Natural products as sources of new drugs from to Lewin, G. Evolution and ecology of actinobacteria and their bioenergy applications. Baltz, R. Marcel Faber Roundtable: is our antibiotic pipeline unproductive because of starvation, constipation or lack of inspiration? Medema, M. A systematic computational analysis of biosynthetic gene cluster evolution: lessons for engineering biosynthesis. PLoS Comput. Doroghazi, J. A roadmap for natural product discovery based on large-scale genomics and metabolomics. Smanski, M. Udwary, D. Genome sequencing reveals complex secondary metabolome in the marine actinomycete Salinispora tropica.

Natl Acad. USA , — Ziemert, N. Diversity and evolution of secondary metabolism in the marine actinomycete genus Salinispora. USA , E—E Jang, K. Anthracimycin, a potent anthrax antibiotic from a marine-derived actinomycete. Schulze, C. Genome-directed lead discovery: biosynthesis, structure elucidation, and biological evaluation of two families of polyene macrolactams against Trypanosoma brucei.


ACS Chem. Ling, L. A new antibiotic kills pathogens without detectable resistance. Pye, C. Retrospective analysis of natural products provides insights for future discovery trends. Miller, I. Interpreting microbial biosynthesis in the genomic age: biological and practical considerations. Drugs 15 , Clardy, J. The natural history of antibiotics. Adnani, N. Coculture of marine invertebrate-associated bacteria and interdisciplinary technologies enable biosynthesis and discovery of a new antibiotic, keyicin.

Zipperer, A. Human commensals producing a novel antibiotic impair pathogen colonization. Oh, D.

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Dentigerumycin: a bacterial mediator of an ant-fungus symbiosis. Currie, C. Ancient tripartite coevolution in the attine ant-microbe symbiosis. VanArnam, E.

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Selvamicin, an atypical antifungal polyene from two alternative genomic contexts. Kroiss, J. Symbiotic streptomycetes provide antibiotic combination prophylaxis for wasp offspring. Scott, J. Bacterial protection of beetle-fungus mutualism. Science , 63—63 Blodgett, J.