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  • Artikel  (640)
  • 2010-2014  (640)
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  • Artikel  (640)
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  • 11
    Publikationsdatum: 2014-11-19
    Beschreibung: The aim of this work was to determine the genetic structure of Rhizobium leguminosarum bv. trifolii population isolated from root nodules of Trifolium repens growing in heavy metal contaminated Bolesław waste-heap area and compare it with that of an unpolluted control Bolestraszyce population. The 684-bp long dinitrogenase reductase ( nifH ) gene fragments were amplified in a PCR reaction and then sequenced. An analysis of nifH gene amplicons of 21 rhizobial strains from each of the studied populations revealed substantially reduced genotype ( h ) and nucleotide ( π ) diversities in the metallicolous Bolesław population in comparison to the non-metallicolous Bolestraszyce one, and showed a significant genetic differentiation between these populations ( F ST  = 0.159, p  = 0.018). Among the strains under investigation, six genotypes (A–F) with 95–99% nifH gene sequence identities were distinguished. Studied T. repens nodule isolates indicated the highest nifH gene sequence similarities (95–100%) with R. leguminosarum bv. trifolii reference strains and on nifH phylogram all these strains formed monophyletic, highly supported clade (100%). The decreased genotype and nucleotide diversities of the waste-heap R. leguminosarum bv. trifolii population, compared to that from the control area and substantial genetic differentiation between populations of nifH gene, is arguably the consequence of the random genetic drift (Tajima's D  = 2.042, p  = 0.99).
    Print ISSN: 0233-111X
    Digitale ISSN: 1521-4028
    Thema: Biologie
    Publiziert von Wiley-Blackwell
    Standort Signatur Einschränkungen Verfügbarkeit
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  • 12
    Publikationsdatum: 2014-11-19
    Beschreibung: An endo-β-1,4-xylanase gene xynA of a thermophilic Geobacillus sp. WBI from “hot” compost was isolated by PCR amplification. The gene encoding 407 residues were overexpressed in E. coli and purified by Ni-NTA chromatography. The purified enzyme (47 kDa) had a broad pH optimum of 6.0 to 9.0, and was active between 50 and 90 °C. The enzyme retained 100% of its activity when incubated at 65 °C for 1 h under alkaline condition (pH 10.0) and retained 75% activity at pH 11.0. The K m and V max of the enzyme were 0.9 mg ml −1 and 0.8 µmol ml −1  min −1 , respectively. In molecular dynamics simulation at 338 K (65 °C), the enzyme was found to be stable. At an elevated temperature (450 K) specific α-helix and β-turns of the proteins were most denatured. The denaturation was less in WBI compared with its highest homolog G. stearothermophilus T-6 xylanase with difference of six residues. The results predict that these regions are responsible for the improved thermostability observed over related enzymes. The present work encourages further experimental demonstration to understand how these regions contribute thermostability to WBI xylanase. The study noted that WBI produces a xylanase with unique characteristics, specifically alkali-thermostability.
    Print ISSN: 0233-111X
    Digitale ISSN: 1521-4028
    Thema: Biologie
    Publiziert von Wiley-Blackwell
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  • 13
    facet.materialart.
    Unbekannt
    Wiley-Blackwell
    Publikationsdatum: 2014-11-14
    Beschreibung: Cover illustration: The leaf-cutter ants are growing fungi on which they feed. For this, they provide the plant biomass to sustain fungal growth. Here, the fungus garden of the dicot-cutting ant (Name kursiv gesetzt) Atta sexdens rubropilosa is shown. the queen and workers can be seen as well. (Photo: Luiz Carlos Forti, UNESP - São Paulo State University, Botucatu, SP, Brazil)
    Print ISSN: 0233-111X
    Digitale ISSN: 1521-4028
    Thema: Biologie
    Publiziert von Wiley-Blackwell
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  • 14
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    Unbekannt
    Wiley-Blackwell
    Publikationsdatum: 2014-11-14
    Print ISSN: 0233-111X
    Digitale ISSN: 1521-4028
    Thema: Biologie
    Publiziert von Wiley-Blackwell
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  • 15
    Publikationsdatum: 2014-11-12
    Beschreibung: Aminopeptidase is an important flavorsome especially in protein hydrolysate debittering by removing hydrophobic amino acid residue at the N-terminal end. Besides, it is also applied to preparation of active peptides and analysis of protein sequence. In this study, leucine aminopeptidase from Bacillus subtilis was cloned and expressed in Pichia pastoris , a widely used heterologous protein expression host. Then it was purified and characterized. After methanol induction for 96 h, the aminopeptidase activity in culture supernatant reached 28.4 U ml −1 , which was 7.1 times that of wild strain B. subtilis Zj016. The optimal temperature and pH of the purified recombinant enzyme were 60 °C and 8.5, respectively. The purified aminopeptidase was stable within 30–60 °C and pH 8.0–9.0. It was intensively inhibited by Ni 2+ , Ca 2+ , DL-dithiothreitol (DTT) and ethylene diamine tetraacetic acid (EDTA), but activated by Co 2+ . The K m toward leucine- p -nitroanilines (Leu- p NA) of the enzyme was 0.97 mM. The sequence analysis of aminopeptidase indicated three potential N -glycosylation sites and it was further verified via MALDI-TOF-MS analysis. Consequently, the N -glycosylated aminopeptidase exhibited higher thermostability and catalytic efficiency. The purified enzyme exhibited two bands through sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS–PAGE) while a single band can be identified when the enzyme was deglycosylated. Circular dichroism spectroscopy indicated that the secondary structure of recombinant aminopeptidase was similar to the wild-type.
    Print ISSN: 0233-111X
    Digitale ISSN: 1521-4028
    Thema: Biologie
    Publiziert von Wiley-Blackwell
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  • 16
    Publikationsdatum: 2014-11-12
    Beschreibung: Lactobacillus sakei is a heterofermentative species of lactic acid bacteria that is used in industrial meat fermentation. To investigate adaptation in a meat environment, whole-genome DNA microarrays were used to analyze the gene expression related to growth and survival of L. sakei strain La22 when grown in sarcoplasmic (S-) or myofibrillar (M-) protein-supplemented chemically defined medium (CDM). Differential expression was detected in 551 genes. Genes encoding enzymes involved in peptide hydrolysis were differentially upregulated in M-CDM or/and S-CDM, and only oppB and oppC , involved in the amino acid and peptide transport system, were upregulated. Most genes related to metabolism of peptides, amino acids and related molecules were over-expressed in M-CDM and S-CDM, except for glnA and metK . Expression of certain genes was according to the differential substrate environment. The expression of genes involved in the stress response was not induced by growth in M-CDM.
    Print ISSN: 0233-111X
    Digitale ISSN: 1521-4028
    Thema: Biologie
    Publiziert von Wiley-Blackwell
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  • 17
    Publikationsdatum: 2014-11-12
    Beschreibung: Many strains of Amycolatopsis , such as Amycolatopsis orientalis, A. balhimycina , and A. mediterranei , are important antibiotic producers. Three indigenous plasmids, pMEA100, pMEA300, and pA387, found in this genus have been sequenced. However, only some vectors based on pA387 have been widely applied in Amycolatopsis research. An indigenous plasmid, pXL100, was isolated from the vancomycin producer A. orientalis HCCB10007. Sequence analysis of pXL100 revealed its total length to be 33,499 bp and GC content to be 68.9%. A 2830-bp fragment containing three ORFs has been identified as essential for replication in A. orientalis , but it has no significant similarity to any known replicons. A vector, pLYZW7-3, was constructed based on the pXL100 replicon and could be transferred into A. mediterranei and A. orientalis by electroporation or conjugation with high frequency. A mutant with a disrupted gene was successfully complemented with the pLYZW7-3 vector, indicating that the vector is potentially useful in Amycolatopsis research.
    Print ISSN: 0233-111X
    Digitale ISSN: 1521-4028
    Thema: Biologie
    Publiziert von Wiley-Blackwell
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  • 18
    Publikationsdatum: 2014-11-12
    Beschreibung: To analyze the pathogenesis at both physiological and molecular level using the model organism, Caenorhabditis elegans at different developmental stages in response to Shigella spp. and its pathogen associated molecular patterns such as lipopolysaccharide. The solid plate and liquid culture-based infection assays revealed that Shigella spp. infects C. elegans and had an impact on the brood size and pharyngeal pumping rate. LPS of Shigella spp. was toxic to C. elegans . qPCR analysis revealed that host innate immune genes have been modulated upon Shigella spp. infections and its LPS challenges. Non-destructive analysis was performed to kinetically assess the alterations in LPS during interaction of Shigella spp. with C. elegans . The modulation of innate immune genes attributed the surrendering of host immune system to Shigella spp. by favoring the infection. LPS appeared to have a major role in Shigella -mediated pathogenesis and Shigella employs a tactic behavior of modifying its LPS content to escape from the recognition of host immune system.
    Print ISSN: 0233-111X
    Digitale ISSN: 1521-4028
    Thema: Biologie
    Publiziert von Wiley-Blackwell
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  • 19
    Publikationsdatum: 2014-10-25
    Beschreibung: Bacillus sonorensis MBCU2 isolated from vermicompost-amended soil from Gujarat, India showed most antagonistic activity against Macrophomina phaseolina by dual culture screening. The culture supernatant of MBCU2 completely suppressed the mycelia growth of pathogen, indicating that suppression was due to the presence of allelochemicals in the culture filtrate. Results of scanning electron microscopy revealed that MBCU2 caused morphological alteration in mycelia of M. phaseolina as evident by hyphal lysis and perforation. Lipopeptides (iturin A and surfactin) produced by MBCU2 were detected and identified by MALDI-TOF-MS as well as liquid chromatography coupled with ESI-MS/MS. Pot trial studies conducted by seed bacterization with MBCU2 resulted in statistically significant increase in Arachis hypogaea L. vegetative growth parameters such as root length (91%), shoot length (252%), fresh weight (71%), dry weight (57%), number of pod (128%), and number of seed (290%) in M. phaseolina infested soil over control as well as decreased M. phaseolina disease severity. We suggest that allelochemicals production can be linked to the mechanism of protection of A. hypogaea L. from M. phaseolina by B. sonorensis MBCU2.
    Print ISSN: 0233-111X
    Digitale ISSN: 1521-4028
    Thema: Biologie
    Publiziert von Wiley-Blackwell
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  • 20
    Publikationsdatum: 2014-10-25
    Beschreibung: Enzymes capable of modifying the sulfated polymeric molecule of fucoidan are mainly produced by different groups of marine organisms: invertebrates, bacteria, and also some fungi. We have discovered and identified a new strain of filamentous fungus Fusarium proliferatum LE1 (deposition number in Russian Collection of Agricultural Microorganisms is RCAM02409), which is a potential producer of fucoidan-degrading enzymes. The strain LE1 (RCAM02409) was identified on the basis of morphological characteristics and analysis of ITS sequences of ribosomal DNA. During submerged cultivation of F. proliferatum LE1 in the nutrient medium containing natural fucoidan sources (the mixture of brown algae Laminaria digitata and Fucus vesiculosus ), enzymic activities of α- L -fucosidase and arylsulfatase were inducible. These enzymes hydrolyzed model substrates, para -nitrophenyl α- L -fucopyranoside and para -nitrophenyl sulfate, respectively. However, the α- L -fucosidase is appeared to be a secreted enzyme while the arylsulfatase was an intracellular one. No detectable fucoidanase activity was found during F. proliferatum LE1 growth in submerged culture or in a static one. Comparative screening for fucoidanase/arylsulfatase/α- L -fucosidase activities among several related Fusarium strains showed a uniqueness of F. proliferatum LE1 to produce arylsulfatase and α- L -fucosidase enzymes. Apart them, the strain was shown to produce other glycoside hydrolyses.
    Print ISSN: 0233-111X
    Digitale ISSN: 1521-4028
    Thema: Biologie
    Publiziert von Wiley-Blackwell
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