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  • 1
    Publication Date: 2016-03-19
    Description: Lignin-derived (e.g. phenolic) compounds can compromise the bioconversion of lignocellulosic biomass to fuels and chemicals due to their toxicity and recalcitrance. The lipid-accumulating bacterium Rhodococcus opacus PD630 has recently emerged as a promising microbial host for lignocellulose conversion to value-added products due to its natural ability to tolerate and utilize phenolics. To gain a better understanding of its phenolic tolerance and utilization mechanisms, we adaptively evolved R. opacus over 40 passages using phenol as its sole carbon source (up to 373% growth improvement over wild-type), and extensively characterized two strains from passages 33 and 40. The two adapted strains showed higher phenol consumption rates (~20 mg/l/h) and ~2-fold higher lipid production from phenol than the wild-type strain. Whole-genome sequencing and comparative transcriptomics identified highly-upregulated degradation pathways and putative transporters for phenol in both adapted strains, highlighting the important linkage between mechanisms of regulated phenol uptake, utilization, and evolved tolerance. Our study shows that the R. opacus mutants are likely to use their transporters to import phenol rather than export them, suggesting a new aromatic tolerance mechanism. The identified tolerance genes and pathways are promising candidates for future metabolic engineering in R. opacus for improved lignin conversion to lipid-based products.
    Print ISSN: 0305-1048
    Electronic ISSN: 1362-4962
    Topics: Biology
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  • 2
    Publication Date: 2012-09-01
    Description: Soil microbiota represent one of the ancient evolutionary origins of antibiotic resistance and have been proposed as a reservoir of resistance genes available for exchange with clinical pathogens. Using a high-throughput functional metagenomic approach in conjunction with a pipeline for the de novo assembly of short-read sequence data from functional selections (termed PARFuMS), we provide evidence for recent exchange of antibiotic resistance genes between environmental bacteria and clinical pathogens. We describe multidrug-resistant soil bacteria containing resistance cassettes against five classes of antibiotics (beta-lactams, aminoglycosides, amphenicols, sulfonamides, and tetracyclines) that have perfect nucleotide identity to genes from diverse human pathogens. This identity encompasses noncoding regions as well as multiple mobilization sequences, offering not only evidence of lateral exchange but also a mechanism by which antibiotic resistance disseminates.〈br /〉〈br /〉〈a href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4070369/" target="_blank"〉〈img src="https://static.pubmed.gov/portal/portal3rc.fcgi/4089621/img/3977009" border="0"〉〈/a〉   〈a href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4070369/" target="_blank"〉This paper as free author manuscript - peer-reviewed and accepted for publication〈/a〉〈br /〉〈br /〉〈span class="detail_caption"〉Notes: 〈/span〉Forsberg, Kevin J -- Reyes, Alejandro -- Wang, Bin -- Selleck, Elizabeth M -- Sommer, Morten O A -- Dantas, Gautam -- T32 GM007067/GM/NIGMS NIH HHS/ -- New York, N.Y. -- Science. 2012 Aug 31;337(6098):1107-11. doi: 10.1126/science.1220761.〈br /〉〈span class="detail_caption"〉Author address: 〈/span〉Center for Genome Sciences and Systems Biology, Washington University School of Medicine, St. Louis, MO 63108, USA.〈br /〉〈span class="detail_caption"〉Record origin:〈/span〉 〈a href="http://www.ncbi.nlm.nih.gov/pubmed/22936781" target="_blank"〉PubMed〈/a〉
    Keywords: Aminoglycosides/pharmacology ; Anti-Bacterial Agents/*pharmacology ; Bacteria/*drug effects/*genetics/pathogenicity ; Base Sequence ; Drug Resistance, Multiple, Bacterial/*genetics ; High-Throughput Screening Assays ; Humans ; Metagenome/*drug effects/*genetics ; Metagenomics ; Molecular Sequence Data ; *Soil Microbiology ; Sulfonamides/pharmacology ; Tetracyclines/pharmacology ; beta-Lactams/pharmacology
    Print ISSN: 0036-8075
    Electronic ISSN: 1095-9203
    Topics: Biology , Chemistry and Pharmacology , Computer Science , Medicine , Natural Sciences in General , Physics
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  • 3
    Publication Date: 2014-05-23
    Description: Ancient and diverse antibiotic resistance genes (ARGs) have previously been identified from soil, including genes identical to those in human pathogens. Despite the apparent overlap between soil and clinical resistomes, factors influencing ARG composition in soil and their movement between genomes and habitats remain largely unknown. General metagenome functions often correlate with the underlying structure of bacterial communities. However, ARGs are proposed to be highly mobile, prompting speculation that resistomes may not correlate with phylogenetic signatures or ecological divisions. To investigate these relationships, we performed functional metagenomic selections for resistance to 18 antibiotics from 18 agricultural and grassland soils. The 2,895 ARGs we discovered were mostly new, and represent all major resistance mechanisms. We demonstrate that distinct soil types harbour distinct resistomes, and that the addition of nitrogen fertilizer strongly influenced soil ARG content. Resistome composition also correlated with microbial phylogenetic and taxonomic structure, both across and within soil types. Consistent with this strong correlation, mobility elements (genes responsible for horizontal gene transfer between bacteria such as transposases and integrases) syntenic with ARGs were rare in soil by comparison with sequenced pathogens, suggesting that ARGs may not transfer between soil bacteria as readily as is observed between human pathogens. Together, our results indicate that bacterial community composition is the primary determinant of soil ARG content, challenging previous hypotheses that horizontal gene transfer effectively decouples resistomes from phylogeny.〈br /〉〈br /〉〈a href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4079543/" target="_blank"〉〈img src="https://static.pubmed.gov/portal/portal3rc.fcgi/4089621/img/3977009" border="0"〉〈/a〉   〈a href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4079543/" target="_blank"〉This paper as free author manuscript - peer-reviewed and accepted for publication〈/a〉〈br /〉〈br /〉〈span class="detail_caption"〉Notes: 〈/span〉Forsberg, Kevin J -- Patel, Sanket -- Gibson, Molly K -- Lauber, Christian L -- Knight, Rob -- Fierer, Noah -- Dantas, Gautam -- DP2 DK098089/DK/NIDDK NIH HHS/ -- DP2-DK-098089/DK/NIDDK NIH HHS/ -- GM 007067/GM/NIGMS NIH HHS/ -- T32 GM007067/GM/NIGMS NIH HHS/ -- T32 HG000045/HG/NHGRI NIH HHS/ -- Howard Hughes Medical Institute/ -- England -- Nature. 2014 May 29;509(7502):612-6. doi: 10.1038/nature13377. Epub 2014 May 21.〈br /〉〈span class="detail_caption"〉Author address: 〈/span〉1] Center for Genome Sciences and Systems Biology, Washington University School of Medicine, St Louis, Missouri 63108, USA [2]. ; 1] Center for Genome Sciences and Systems Biology, Washington University School of Medicine, St Louis, Missouri 63108, USA [2] Department of Pathology and Immunology, Washington University School of Medicine, St Louis, Missouri 63110, USA [3]. ; Center for Genome Sciences and Systems Biology, Washington University School of Medicine, St Louis, Missouri 63108, USA. ; Cooperative Institute for Research in Environmental Sciences, University of Colorado, Boulder, Colorado 80309, USA. ; 1] Department of Chemistry and Biochemistry and BioFrontiers Institute, University of Colorado, Boulder, Colorado 80309, USA [2] Howard Hughes Medical Institute, Boulder, Colorado 80309, USA. ; 1] Cooperative Institute for Research in Environmental Sciences, University of Colorado, Boulder, Colorado 80309, USA [2] Department of Ecology and Evolutionary Biology, University of Colorado, Boulder, Colorado 80309, USA. ; 1] Center for Genome Sciences and Systems Biology, Washington University School of Medicine, St Louis, Missouri 63108, USA [2] Department of Pathology and Immunology, Washington University School of Medicine, St Louis, Missouri 63110, USA [3] Department of Biomedical Engineering, Washington University, St Louis, Missouri 63130, USA.〈br /〉〈span class="detail_caption"〉Record origin:〈/span〉 〈a href="http://www.ncbi.nlm.nih.gov/pubmed/24847883" target="_blank"〉PubMed〈/a〉
    Keywords: Agriculture ; Anti-Bacterial Agents/pharmacology ; Bacteria/classification/drug effects/*genetics/*isolation & purification ; Drug Resistance, Microbial/drug effects/*genetics ; *Ecosystem ; Fertilizers ; Gene Transfer, Horizontal/genetics ; Genes, Bacterial/drug effects/genetics ; Genome, Bacterial/drug effects/genetics ; Integrases/genetics ; Metagenome/drug effects/*genetics ; Metagenomics ; Models, Genetic ; Molecular Sequence Data ; Nitrogen/metabolism/pharmacology ; Open Reading Frames/genetics ; *Phylogeny ; Poaceae/growth & development ; RNA, Ribosomal, 16S/genetics ; *Soil Microbiology ; Synteny/genetics ; Transposases/genetics
    Print ISSN: 0028-0836
    Electronic ISSN: 1476-4687
    Topics: Biology , Chemistry and Pharmacology , Medicine , Natural Sciences in General , Physics
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  • 4
    Publication Date: 2005-11-27
    Description: Assessment of techniques for determining eigenvalues of large systems
    Keywords: STRUCTURAL MECHANICS
    Type: NASA. LEWIS RES. CENTER SPACE TRANSPORTATION SYSTEM TECHNOL. SYMP., VOL. 2 JUL. 1970; P 4-27
    Format: text
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  • 5
    Publication Date: 2014-09-09
    Description: Silica reusable surface insulation is analyzed for its cold soak properties, bond properties at low temperature, gap and step heating effects, and aerodynamic surface roughness effects to determine space shuttle weight and cost sensitivity to structure subsystem weights. Computerized simulation studies show that total program cost savings of silica (vs. mullite) exceed thermal protection system design and development costs and that an early introduction of silica increases total program cost savings.
    Keywords: MATERIALS, NONMETALLIC
    Type: NASA. Ames Res. Center Symp. on Reusable Surface Insulation for Space Shuttle, Vol. 3; p 1121-1183
    Format: text
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