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  • Articles  (26,588)
  • Blackwell Publishing Ltd  (26,588)
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  • Articles  (26,588)
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  • 1
    Electronic Resource
    Electronic Resource
    Evidence for past integration of IncP-1 plasmids into bacterial chromosomes (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 241 (2004), S. 0 
    Details
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Plasmids of the IncP-1 incompatibility group are self-transmissible between and stably maintained in a very broad range of Gram-negative bacteria. A characteristic feature of IncP-1 genomes is the existence of multiple binding sites (OB) for the KorB protein which plays a dual role in active partitioning of plasmid and coordinate regulation of expression of genes for replication, maintenance and transfer. A search of the available bacterial genome sequences revealed a significant number (70 out of 322) with one or more putative KorB binding sites. Binding of KorB to such a site was demonstrated by chromatin immunoprecipitation (ChIP) for Pseudomonas putida KT2440. While such a site may arise by chance, this is unlikely for Pseudomonas aeruginosa UCBPP-PA14 whose genome sequence contains four clustered OB sites and several regions have more than 80% nucleotide identity to traJ, trbJ and trbL of IncP-1 plasmids. A number of other bacterial genomes also contain integrated partial IncP-1 genomes or their remnants. These data provide evidence for multiple past integration events of IncP-1 plasmids into bacterial chromosomes and provide new evidence for IncP-1 plasmids being important elements in gene mobility.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1016/j.femsle.2004.10.016
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  • 2
    Electronic Resource
    Electronic Resource
    Inhibition of methicillin-resistant Staphylococcus aureus by an in vitro continuous-flow culture containing human stool microflora (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 241 (2004), S. 0 
    Details
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: We used an in vitro continuous-flow culture model of human stool microflora to examine the ability of human stool microflora to inhibit growth of two methicillin-resistant S. aureus (MRSA) strains. Continuous-flow cultures consistently eliminated MRSA inocula of 106 cfu/mL within 4 days, and addition of continuous-flow culture resulted in elimination of a pre-established MRSA culture (∼108 cfu/mL) within 6–8 days. Anaerobic or “aerobic” (i.e., continuous bubbling of room air to eliminate obligate anaerobes) cultures eliminated MRSA at similar rates. The MRSA strains were unable to replicate under anaerobic conditions in sterile filtrates produced from the continuous-flow culture, but rapid growth occurred when glucose was added. These data demonstrate that indigenous stool microflora efficiently eliminate MRSA colonization and obligate anaerobes are not essential for inhibition. Our findings also suggest that inhibition of MRSA in continuous-flow cultures is due to depletion of nutrients rather than production of inhibitory conditions.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1016/j.femsle.2004.10.021
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  • 3
    Electronic Resource
    Electronic Resource
    Non-exhaustive extraction techniques (NEETs) for the prediction of naphthalene mineralisation in soil (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 241 (2004), S. 0 
    Details
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Non-exhaustive extraction techniques (NEETs) have been shown to measure the putatively bioavailable fraction of hydrophobic compounds in soil. To date, these studies have only considered bioavailability in a single soil type. In this study, naphthalene was amended into five different soil types and mineralisation, bacterial biosensor response and the number of indigenous microbial naphthalene degraders were determined. Two NEETs were used to extract the naphthalene from soil; hydroxypropyl-β-cyclodextrin (HPCD) and XAD-4. The HPCD extractable fraction correlated closely (R2= 0.917) with the portion that was mineralised, but the XAD-4 extract did not (R2= 0.044). HPCD may be ideal for the rapid assessment of the fraction of a hydrophobic organic contaminant that is available for biodegradation. A NEET that complements environmental microbial analysis will enhance our understanding of soil pollution interactions and equip us better in designing risk assessment models that integrate biological parameters. This application, although refined for soil samples, should be transferable to other environmental matrices.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1016/j.femsle.2004.10.023
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  • 4
    Electronic Resource
    Electronic Resource
    Differential gene expression during interactions between Heterobasidion annosum and Physisporinus sanguinolentus (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 241 (2004), S. 0 
    Details
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Using mRNA differential display we have identified differentially expressed genes in non-self-interacting vs. single mycelia of the conifer pathogen Heterobasidion annosum and the wood decomposing basidiomycete Physisporinus sanguinolentus. Altogether 39 differentially displayed bands were cloned and sequenced, corresponding to 21 unique genes, which were confirmed by semi-quantitative RT-PCR to be differentially expressed. Further confirmation of differential gene expression was made by real time RT-PCR. All 10 genes identified from P. sanguinolentus had lower expression, while in H. annosum three genes had higher and eight lower expression in non-self-interacting mycelia vs. single mycelia. One of the induced genes showed high similarity to the Coprinus cinereus recA/RAD51 homolog (rah1) which is essential for homologous recombination, DNA repair and stress responses.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1016/j.femsle.2004.10.007
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  • 5
    Electronic Resource
    Electronic Resource
    Rapid detection of Mycoplasma dispar and M. bovirhinis using allele specific polymerase chain reaction protocols (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 241 (2004), S. 0 
    Details
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: We describe an allele specific PCR based approach for the rapid detection of two bovine Mycoplasma species associated with respiratory disease. Specific and universal oligonucleotides were used in combination to detect the presence of single nucleotide polymorphisms within the 16S ribosomal DNA sequence. Presence of Mycoplasma 16S rDNA is indicated by the production of a single control fragment, whilst positive samples generate an alternative smaller specific product over the same region. This technique provides a reliable and sensitive method which, although widely used in human genetic screening, has not been documented for diagnosis of bacterial infection.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1016/j.femsle.2004.10.010
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  • 6
    Electronic Resource
    Electronic Resource
    Mitochondrial DNA organisation of the mtDNA type 2b of Aspergillus tubingensis compared to the Aspergillus niger mtDNA type 1a (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 241 (2004), S. 0 
    Details
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: A physical and functional map of Aspergillus tubingensis mtDNA type 2b was constructed and compared to Aspergillus niger type 1a mtDNA. The gene content and order, as well as the patterns of restriction sites, were similar. Two unidentified ORFs and several repeat elements were found in the region between the ndh1 and ndh4 genes on both mtDNAs. The sizes of the A. niger and A. tubingensis mtDNAs were 31.23 and 33.06 kb, respectively, the difference was principally attributed to the altered intron content of their cox1 genes.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1016/j.femsle.2004.10.025
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  • 7
    Electronic Resource
    Electronic Resource
    Release of transforming plasmid DNA from actively growing genetically engineered Escherichia coli (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 240 (2004), S. 0 
    Details
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: We studied the transforming ability of the extracellular plasmid DNA released from a genetically engineered Escherichia coli pEGFP and the culturing conditions for the release of transforming DNA. The transforming ability was evaluated by transformation of competent cells with filtrates of E. coli pEGFP cultures. The number of transformants increased with time when E. coli pEGFP cells grew exponentially in rich medium, but not in stationary phase or when inoculated in freshwater. These results suggested that crude extracellular plasmid DNA had transforming ability and this transforming DNA was mainly released by actively growing bacteria.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1016/j.femsle.2004.09.024
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  • 8
    Electronic Resource
    Electronic Resource
    Molecular hydrogen from water radiolysis as an energy source for bacterial growth in a basin containing irradiating waste (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 240 (2004), S. 0 
    Details
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Although being deionized, filtered and therefore normally deeply oligotrophic, the water from a basin containing irradiating waste presented relatively high bacterial concentrations (ca 105 cfu ml−1) and biofilm development at its surface and on the walls. This water was characterized by a high concentration of molecular H2 due to water radiolysis, while its electrochemical potential was around +400 mV due the presence of dissolved O2 and active oxygen compounds. This combination of H2 availability and of an oxidant environment is completely original and not described in nature. From surface and wall biofilms, we enumerated the autotrophic populations (∼105 bacteria ml−1) able to grow in presence of H2 as energy source and CO2 as carbon source, and we isolated the most abundant ones among cultivable bacteria. They efficiently grew on a mineral medium, in the presence of H2, O2 and CO2, the presence of the three gases being indispensable. Two strains were selected and identified using their rrs gene sequence as Ralstonia sp. GGLH002 and Burkholderia sp. GGLH005. In pure culture and using isotope exchange between hydrogen and deuterium, we demonstrated that these strains are able to oxidize hydrogen as energy source, using oxygen as an electron acceptor, and to use carbon dioxide as carbon source. These chemoautotroph hydrogen-oxidizing bacteria probably represent the pioneer bacterial populations in this basin and could be primary producers in the bacterial community.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1016/j.femsle.2004.09.025
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  • 9
    Electronic Resource
    Electronic Resource
    Genetic diversity of small eukaryotes from the coastal waters of Nansha Islands in China (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 240 (2004), S. 0 
    Details
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Population structures and genetic diversity of the small eukaryotic plankton from the coastal waters of the Nansha Islands in China were investigated. Two genes libraries using 18S rDNA of the marine small eukaryotes were constituted, and 323 clones were identified within alveolates (more than 43%), acanthareas, viridiplantaes, and stramenopiles. Many novel clones were detected in the two libraries, including two groups of alveolates and two clades related to both acanthareas and polycystineas. Several sequences unrelated to any other known eukaryotes may represent early branches in the phylogenetic tree. Our results reveal that there is a high diversity and abundance of small eukaryotes in the marine regions of China.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1016/j.femsle.2004.09.030
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  • 10
    Electronic Resource
    Electronic Resource
    Single mutations of residues outside the active center of the xylanase Xys1Δ from Streptomyces halstedii JM8 affect its activity (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 240 (2004), S. 0 
    Details
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Mutagenesis of the xylanase Xys1 of Streptomyces halstedii JM8 has been done by error prone PCR. Mutants with modified hydrolytic activity were isolated, the recombinant variant proteins purified and the catalytic activities of each one determined and compared with the wild type enzyme. Two of the isolated single point mutants, m1 (G133D) and m8 (N148D), showed 22–25% increase in specific activity towards xylan compared to wild type xylanase. Two other mutants, m5a (D175A) and m7 (T160A), showed a significant reduction in specific activity of 40–50% with respect to the wild type enzyme. These residues are mainly located in the βα-loops of the xylanase, the region showing the main structural divergences within family 10 of xylanases. This study shows the usefulness of random mutagenesis to point out some key residues not directly involved in the active center, but in which mutation produces subtle structural rearrangements affecting the enzymatic function.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1016/j.femsle.2004.09.032
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  • 11
    Electronic Resource
    Electronic Resource
    A divIVA null mutant of Staphylococcus aureus undergoes normal cell division (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 240 (2004), S. 0 
    Details
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: DivIVA is involved in placement of the division septum and chromosome segregation in Bacillus subtilis and it plays important roles in cell division or morphogenesis in diverse Gram-positive bacteria. In Staphylococcus aureus, DivIVA is localized at the division septum, but it does not colocalize with the chromosomal origin of replication, as labeled with SpoOJ protein. Unexpectedly, a divIVA null mutant is not impaired in growth, nor is it affected in chromosome segregation or cell morphology.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1016/j.femsle.2004.09.038
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  • 12
    Electronic Resource
    Electronic Resource
    Characterization of DegU, a response regulator in Listeria monocytogenes, involved in regulation of motility and contributes to virulence (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 240 (2004), S. 0 
    Details
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: The degU (lmo2515) gene encodes a putative response regulator in the food-borne pathogen Listeria monocytogenes. It has 63% amino acid identity to the DegU response regulator of Bacillus subtilis. We have characterized the degU gene product in L. monocytogenes EGD by generation of a deletion mutant. The ΔdegU mutant was found to be non-motile in motility plate assay and no flagellin was detected. The mutant was attenuated in challenge of mice. Northern blot analysis suggested that the degU gene product is a transcriptional activator of the flagellin gene, flaA, at 25 °C. However, the degU gene product had no influence on the transcription of prfA encoding the major virulence regulator, PrfA. The results indicate that the putative DegU response regulator is a pleiotropic regulator involved in expression of both motility at low temperature and in vivo virulence in mice.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1016/j.femsle.2004.09.039
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  • 13
    Electronic Resource
    Electronic Resource
    Electron microscopy analysis of Mycobacterium tuberculosis cell division (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 240 (2004), S. 0 
    Details
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: The ultrastructure of Mycobacterium tuberculosis cells undergoing division was examined by electron microscopy. Two features of cell division were observed and are described here. First, cells are capable of undergoing a type of “snapping” postfission movement. This movement is likely due to a multi-layered cell wall in which the inner layer participates in septum formation while the outer layer ruptures first on one side. A second feature related to cell division is the ability of dividing cells to form transient branching structures.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1016/j.femsle.2004.09.004
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  • 14
    Electronic Resource
    Electronic Resource
    Multicellular life cycle of magnetotactic prokaryotes (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 240 (2004), S. 0 
    Details
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Most multicellular organisms, prokaryotes as well as animals, plants, and algae have a unicellular stage in their life cycle. Here, we describe an uncultured prokaryotic magnetotactic multicellular organism that reproduces by binary fission. It is multicellular in all the stages of its life cycle, and during most of the life cycle the cells organize into a hollow sphere formed by a functionally coordinated and polarized single-cell layer that grows by increasing the cell size. Subsequently, all the cells divide synchronously; the organism becomes elliptical, and separates into two equal spheres with a torsional movement in the equatorial plane. Unicellular bacteria similar to the cells that compose these organisms have not been found. Molecular biology analysis showed that all the organisms studied belong to a single genetic population phylogenetically related to many-celled magnetotactic prokaryotes in the delta sub-group of the proteobacteria. This appears to be the first report of a multicellular prokaryotic organism that proliferates by dividing into two equal multicellular organisms each similar to the parent one.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1016/j.femsle.2004.09.035
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  • 15
    Electronic Resource
    Electronic Resource
    Microbial degradation of the organophosphate pesticide, Ethion (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 240 (2004), S. 0 
    Details
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: The organophosphate pesticide, Ethion, remains a major environmental contaminant in rural Australia and poses a significant threat to environmental and public health. The aerobic degradation of Ethion by mesophilic bacteria isolated from contaminated soils surrounding disused cattle dip sites was investigated. Two isolates, identified as Pseudomonas and Azospirillum species, were capable of biodegrading Ethion when cultivated in minimal salts medium. The abiotic hydrolytic degradation products of Ethion such as Ethion Dioxon and O,O-diethylthiosphosphate were not detected. The data suggest the rapid degradation of Ethion to support microbial growth. The results have implications for the development of a bioremediation strategy.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1016/j.femsle.2004.09.010
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  • 16
    Electronic Resource
    Electronic Resource
    Single channel evidence for innate pore-formation by Vibrio parahaemolyticus thermostable direct haemolysin (TDH) in phospholipid bilayers (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 240 (2004), S. 0 
    Details
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Vibrio parahaemolyticus thermostable direct haemolysin (TDH) is widely considered to be a pore-forming toxin. The protein has no significant homology to other known pore-forming toxins and its mechanism of action in vivo remains undefined. We demonstrate single channel pore-forming activity of V. parahaemolyticus TDH in planar lipid bilayers. Channel conductance ranged between 30–450 pS in 0.5 M KCl with a calculated cation selectivity (PK/PCl) of 2.7. Channels were formed in NaCl and choline-Cl with and without cholesterol present and in the presence of neutral or negatively charged phospholipids. Zinc ions did not block pore formation. Whilst various techniques have previously suggested that TDH is a pore-forming toxin, the data in this study provide direct single channel evidence and indicate several features of pore formation in synthetic phospholipid membranes.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1016/j.femsle.2004.09.015
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  • 17
    Electronic Resource
    Electronic Resource
    Enzymatic reactions in anaerobic 2-methylnaphthalene degradation by the sulphate-reducing enrichment culture N 47 (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 240 (2004), S. 0 
    Details
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: The upper pathway of anaerobic degradation of 2-methylnaphthalene was studied with a sulphate-reducing enrichment culture, which is able to grow with naphthalene or 2-methylnaphthalene as sole carbon source and electron donor. Anaerobic degradation of 2-methylnaphthalene is initiated by an addition of fumarate to the methyl-group producing the first intermediate, naphthyl-2-methyl-succinate. In a subsequent β-oxidation of the original methyl atom, the central metabolite 2-naphthoic acid is generated. In the following pathway, the aromatic ring system is reduced, cleaved, and finally oxidised to CO2. Here, we present two new enzymatic reactions of the 2-methylnaphthalene degradation pathway that were measured in crude cell extracts. All metabolites were identified with HPLC by co-elution with synthesised reference substances. The first enzyme, succinyl-CoA:naphthyl-2-methyl-succinate CoA-transferase, catalyses the activation of naphthyl-2-methyl-succinic acid to the corresponding CoA ester. The average specific activity of this enzyme was 19.6 nmol × min−1× mg of protein−1. The CoA-transfer was not inhibited by sodium borohydride and only partially by hydroxylamine, indicating that this enzyme belongs to the family III of CoA-transferases like the corresponding enzyme in the anaerobic toluene degradation pathway. The product of this CoA-transfer reaction, naphthyl-2-methyl-succinyl-CoA is then oxidised in a reaction to naphthyl-2-methylene-succinyl-CoA by the enzyme naphthyl-2-methyl-succinyl-CoA dehydrogenase. The specific activity of this enzyme was 0.115 nmol × min−1× mg of protein−1. The enzymatic activity could only be detected using phenazine methosulphate as electron acceptor. No activity was observed with natural electron acceptors such as nicotinamide adenine dinucleotide or flavin adenine dinucleotide. The two novel reactions presented here demonstrate that the original methyl-group of 2-methylnaphthalene is oxidised to the carboxyl group of 2-naphthoic acid in the upper part of the anaerobic degradation pathway.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1016/j.femsle.2004.09.014
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  • 18
    Electronic Resource
    Electronic Resource
    Glycopeptide resistance determinants from the teicoplanin producer Actinoplanes teichomyceticus (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 240 (2004), S. 0 
    Details
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: In enterococci and other pathogenic bacteria, high-level resistance to vancomycin and other glycopeptide antibiotics requires the action of the van genes, which direct the synthesis of peptidoglycan terminating in the depsipeptide d-alanyl-d-lactate, in place of the usual d-Ala-d-Ala. The Actinoplanes teichomyceticus tcp cluster, devoted to the biosynthesis of the glycopeptide antibiotic teicoplanin, contains van genes associated to a murF-like sequence (murF2). We show that A. teichomyceticus contains also a house-keeping murF1 gene, capable of complementing a temperature sensitive Escherichia coli murF mutant. MurF1, expressed in Streptomyces lividans, can catalyze the addition of either d-Ala-d-Ala or d-Ala-d-Lac to the UDP-N-acetyl-muramyl-l-Ala-d-Glu-d-Lys. However, similarly expressed MurF2 shows a small enzymatic activity only with d-Ala-d-lactate. Introduction of a single copy of the entire set of van genes confers resistance to teicoplanin-type glycopeptides to S. coelicolor.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1016/j.femsle.2004.09.017
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  • 19
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    An endo-polygalacturonase (PG) of Fusarium moniliforme escaping inhibition by plant polygalacturonase-inhibiting proteins (PGIPs) provides new insights into the PG–PGIP interaction (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 240 (2004), S. 0 
    Details
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Polygalacturonase-inhibiting proteins (PGIPs) are plant defence molecules inhibiting the activity of fungal endo-polygalacturonases (endo-PGs). We found that soybean and bean PGIPs inhibited the endo-PG activity produced by the isolate FC-10 of Fusarium moniliforme but not the enzyme activity produced by the isolate PD of F. moniliforme. The bean PGIP proved to be ineffective against all the PG isoforms produced by the PD isolate. Deduced amino acid sequence comparison between PGs from PD, FC-10 and 62264 isolates identified the structural regions of the enzyme possibly related to its resistance to PGIP inhibition. These include one region at the N-terminal portion of the enzyme and a few single amino acid substitutions along the entire sequence, two of which surrounding the active site.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1016/j.femsle.2004.09.019
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  • 20
    Electronic Resource
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    S-Nitrosoglutathione cytotoxicity to Mycobacterium smegmatis and its use to isolate stationary phase survival mutants (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 239 (2004), S. 0 
    Details
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: We report that stationary phase Mycobacterium smegmatis is more sensitive than exponential phase cells to the nitric oxide donor S-Nitrosoglutathione (GSNO). This finding was used to select for both spontaneous and transposon mutants of M. smegmatis with increased resistance to GSNO in stationary phase. Some of these mutants were also defective in stationary phase survival, demonstrating a link between sensitivity to GSNO and stationary phase survival. Transduction of the disrupted region from seven selected mutants indicated that the transposon insertion was linked to the GSNO-resistance and stationary phase survival phenotypes. For five mutants, the disrupted genes were identified. Three were homologous to genes with possible roles in nutrient scavenging, including: (i) a putative amino acid efflux pump, (ii) a putative thioesterase and (iii) an enoyl-CoA-hydratase. One mutant was disrupted in the atpD gene, encoding the β chain of F1 F0 ATP synthase. We independently isolated a stationary phase survival mutant disrupted in the atpA gene (encoding the α chain) of the F1 F0 ATP synthase of the same operon, suggesting an important role for efficient ATP synthesis in stationary phase survival.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1016/j.femsle.2004.08.030
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  • 21
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    Regulation system for protease production in Vibrio vulnificus (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 240 (2004), S. 0 
    Details
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Vibrio vulnificus is a causative agent of serious food-borne diseases in humans related to consumption of raw seafoods. This human pathogen secretes a metalloprotease (VVP) that evokes enhancement of the vascular permeability and disruption of the capillaries. Production of microbial proteases is generally induced at early stationary phase of its growth. This cell density dependent regulation of VVP production in V. vulnificus known to be the quorum-sensing. When V. vulnificus was cultivated in Luria–Bertani (LB) medium, accumulation of the autoinducer, the signal molecule operating the quorum-sensing system, was detected. Moreover, expression of the vvp gene encoding VVP was found to be closely related with expression of the luxS gene that encode the synthase of the autoinducer precursor (luxS). These findings may indicate VVP production is controlled by the quorum-sensing system in LB medium. Futhermore, this system functioned more effectively at 26 °C than at 37 °C. When incubated at 37 °C in human serum supplemented with ferric chloride, production of VVP and expression of vvp increased in proportion to the concentration of ferric ion; whereas, expression of luxS was not increased. This suggests that VVP production in human serum containing ferric ion may be regulated mainly by the system other than the quorum-sensing system.
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    Subject index volume 238 (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 238 (2004), S. 0 
    Details
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
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    URL: http://dx.doi.org/10.1111/j.1574-6968.2004.tb09792.x
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    Insertional mutagenesis and characterization of a polyketide synthase gene (PKS1) required for melanin biosynthesis in Bipolaris oryzae (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 238 (2004), S. 0 
    Details
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: A polyketide synthase gene named PKS1, involved in the melanin biosynthesis pathway of the phytopathogenic fungus Bipolaris oryzae, was isolated using restriction enzyme-mediated integration. Sequence analysis showed that the PKS1 encodes a putative protein that has 2155 amino acids and significant similarity to other fungal polyketide synthases. Targeted disruption of the PKS1 gene showed that it is necessary for melanin biosynthesis in B. oryzae. Northern blot analysis showed that PKS1 transcripts were specifically enhanced by near-ultraviolet radiation (300–400 nm) and that its temporal transcriptional patterns were similar to those of THR1 and SCD1 genes involved in the melanin biosynthesis pathway of B. oryzae.
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    Identification, characterisation and specificity of a cell wall lytic enzyme from Lactobacillus fermentum BR11 (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 238 (2004), S. 0 
    Details
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Screening of a genomic library with an antiserum raised against whole Lactobacillus fermentum BR11 cells identified a clone expressing an immunoreactive 37-kDa protein. Analysis of the 3010-bp DNA insert contained within the clone revealed four open reading frames (ORFs). One ORF encodes LysA, a 303 amino acid protein which has up to 35% identity with putative endolysins from prophages Lj928 and Lj965 from Lactobacillus johnsonii and Lp1 and Lp2 from Lactobacillus plantarum as well as with the endolysin of Lactobacillus gasseri bacteriophage Φadh. The immunoreactive protein was shown to be encoded by a truncated ORF downstream of lysA which has similarity to glutamyl-tRNA synthetases. The N-terminus of LysA has sequence similarity with N-acetylmuramidase catalytic domains while the C-terminus has sequence similarity with putative cell envelope binding bacterial SH3b domains. C-terminal bacterial SH3b domains were identified in the majority of Lactobacillus bacteriophage endolysins. LysA was expressed in Escherichia coli and unusually was found to have a broad bacteriolytic activity range with activity against a number of different Lactobacillus species and against Lactococcus lactis, streptococci and Staphylococcus aureus. It was found that LysA is 2 and 8000 times more active against L. fermentum than L. lactis and Streptococcus pyogenes, respectively.
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    Rapid identification of pickle yeasts by fluorescent PCR and microtemperature-gradient gel electrophoresis (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 238 (2004), S. 0 
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    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Monitoring the yeast populations within pickle soaking fluid is imperative for ensuring optimum taste, but these analyses have proven time-consuming and expensive, limiting their industrial application. Here, yeasts were identified in the soaking fluid from Japanese radish pickles using fluorescent PCR amplification of the variable D1/D2 region of the 26S rDNA, followed by analysis with microtemperature-gradient gel electrophoresis (μ-TGGE). This smaller version of the normal TGGE apparatus is capable of analyzing samples 10- to 20-fold faster without sacrificing data quality. Each primer set was labeled with a different fluorescent dye, allowing easy isolation of the various PCR products and identification of the bands corresponding to the various yeasts. The results indicate that fluorescent PCR and μ-TGGE may be a useful new method for rapid, easy monitoring of yeast flora in various food industries. This new method can be used on a daily basis to provide overviews of yeast flora during pickle production, allowing producers to quickly grasp pickle readiness at a single glance.
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    The N-terminal family 22 carbohydrate-binding module of xylanase 10B of Clostridium themocellum is not a thermostabilizing domain (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 238 (2004), S. 0 
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    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Xylanase Xyn10B from Clostridium thermocellum is a modular enzyme that contains two family 22 carbohydrate binding modules N- (CBM22-1) and C- (CBM22-2) terminal of the family 10 glycoside hydrolase catalytic domain (GH10). In a previous study, we showed that removal of CBM22-1 reduces the resistance to thermoinactivation of the enzyme suggesting that this module is a thermostabilizing domain. Here, we show that it is the module border on the N-terminal side of GH10 that confers resistance to thermoinactivation and to proteolysis. Therefore, CBM22-1 does not function as a thermostabilizing domain and the role for this apparently non-functional CBM remains elusive.
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    Influence of ethyl acetate production and ploidy on the anti-mould activity of Pichia anomala (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 238 (2004), S. 0 
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    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: A diploid and a haploid strain of Pichia anomala were tested for their biocontrol ability against the spoilage mould Penicillium roqueforti in glass tubes filled with grain at two water activities (aw). At aw 0.98, the two yeast strains grew and inhibited mould growth equally well and showed similar patterns of ethyl acetate production, reaching maximum values of 10–14 μg ml−1 headspace. At aw 0.95, both growth and biocontrol performance of the haploid strain were reduced. Ethyl acetate formation was also substantially reduced, with maximum headspace concentrations of 4 μg ml−1. We conclude that ethyl acetate is a major component of the anti-mould activity. The inhibitory effect of ethyl acetate was confirmed in a bioassay where the pure compound reduced biomass production of P. roqueforti.
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    Genetic diversity (RAPD-PCR) of lactobacilli isolated from “Almagro” eggplant fermentations from two seasons (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 238 (2004), S. 0 
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    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Genetic diversity of 323 strains of lactobacilli isolated from an Almagro eggplant manufacturing plant was analyzed by using random amplified polymorphic DNA (RAPD). Thirty-four distinct RAPD patterns were obtained with 95% of isolates grouped into 18 main clusters. Genetic diversity was higher in brines from season II, Lactobacillus plantarum and Lactobacillus fermentum/cellobiosus being the species with the greatest number of genotypes. A single L. fermentum/cellobiosus genotype comprised isolates from both seasons and could be considered endemic to that factory.
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    Involvement of cyclic AMP in the production of the acid protease Acp1 by Sclerotinia sclerotiorum (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 237 (2004), S. 0 
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    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: The acid protease Acp1 is produced by Sclerotinia sclerotiorum during plant infection. We explored the mechanism involved in the triggering of that production and found that cyclic AMP played a positive role. Acp1 could be produced in the sole presence of exogenous cyclic AMP. The use of molecules known to increase or decrease the intracellular cyclic AMP levels confirmed the impact of this nucleotide on the protease production and suggested its endogenous site of action. Further pharmacological studies showed the specific effect of cyclic AMP on Acp1 production and suggested that protein kinase A would be its likely target. Together, these results provide the first indication that the production of a pathogenesis-related fungal protease could depend on a cyclic AMP/Protein kinase A signalling pathway.
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    Stress responses of linear plasmids from Debaryomyces hansenii (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 237 (2004), S. 0 
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    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: The triplet linear plasmids pDHL1/2/3 from the salt-tolerant yeast Debaryomyces hansenii TK are localized in the cytoplasm and characterized by a unique feature that they require environmental stressors (0.3 M NaCl or solutes such as sorbitol with equivalent osmolarity) for stable replication and maintenance. The degree of osmolarity dependence of pDHLs was greatly affected by growth temperature of the host cells: the stability of pDHLs was maintained in the absence of osmolarity in cells growing at 25 °C, and required osmorarity equivalent to 0.3–1.0 M NaCl on shifting to 30–35 °C. Although to less extent, similar osmolarity dependence at high temperatures was observed with another system of D. hansenii linear plasmids. Short-term conditioning of cells to heat or high osmolarity resulted in significant improvement in the plasmid stability, suggesting possible involvement of stress proteins and/or high glycerol level in the stabilization process.
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    Evaluation of polygalacturonase activity in Saccharomyces cerevisiae wine strains (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 237 (2004), S. 0 
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    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: A total of 61 S. cerevisiae strains, 60 of them isolated from wine ecosystems, were evaluated for the presence of the gene encoding endopolygalacturonase (PGU1) and for polygalacturonase (PG) activity. Nine strains lack the gene PGU1 and did not exhibit PG activity on plate assays. Of the 52 strains showing an amplified band corresponding to the size of PGU1 gene, only 36 degraded polygalacturonic acid (PGA) and 17 did not degrade it at any of the pH values used. The coding region of the PGU1 gene (ORF YJR153w) was not present in some PG activity negative strains. The S. cerevisiae UCLMS-39 strain was selected for its specific activity at different pHs, temperatures and oenological parameters. The temperature and pH optima were 50 °C and 3.5–5.5 respectively and it was only affected by ethanol. The PGU1 gene was cloned and sequenced. The production of a biologically functional endoPG in S. cerevisiae UCLMS-39 brings us a step closer to improving the qualities of outstanding enological yeasts naturally lacking PG activity.
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    Expression of the Staphylococcus aureus surface proteins HtrA1 and HtrA2 in Lactococcus lactis (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 237 (2004), S. 0 
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    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Staphylococcus aureus encodes two HtrA-like serine surface proteases, called HtrA1 and HtrA2. The roles of these HtrA homologs were distinguished by expression studies in a heterologous host, Lactococcus lactis, whose single extracellular protease, HtrALl, was absent. HtrALl is involved in stress resistance, and processing and/or degradation of extracellular proteins. Controlled expression of staphylococcal htrA1 and htrA2 was achieved in L. lactis strain NZ9000 ΔhtrA, as confirmed with anti-HtrA1 and anti-HtrA2 specific antibodies. HtrA1 fully restored thermo-resistance to the htrA-defective L. lactis strain, despite a poor capacity to degrade abnormal or truncated proteins. We therefore propose that activities of HtrA1 other than proteolysis may be sufficient for high-temperature growth complementation. HtrA2 is 36% identical to HtrALl, and was highly expressed in L. lactis; nevertheless, it displayed nearly no detectable activities. The poor proteolytic activities of staphylococcal HtrA proteins in L. lactis may reflect a requirement for S. aureus-specific co-factors.
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    A novel glutamate-dependent acid resistance among strains belonging to the Proteeae tribe of Enterobacteriaceae (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 237 (2004), S. 0 
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    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Morganella, Providencia and Proteus strains were capable of surviving pH 2.0 for 1 h if glutamate was present. These strains did not have glutamic acid decarboxylase activity and the gadAB genes were not detected in any of these bacteria. When exposed to pH 2.0 acid shocks, the survival rate of these bacteria was significantly increased with glutamate concentrations as low as 0.3 mM in the acid media. Escherichia coli cells incubated at pH 3.4 consumed four times more glutamate and produced at least 7-fold more γ-amino butyric acid than Morganella, Providencia and Proteus strains. These results indicate that strains belonging to the Proteeae tribe might have novel glutamate dependent acid-resistance mechanisms.
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    Hydrogen peroxide concentrations detected in Agaricus bisporus sporocarps and relation with their susceptibility to the pathogen Verticillium fungicola (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 237 (2004), S. 0 
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    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: A dry bubble is an undifferentiated structure that forms in place of mushrooms when cultures of Agaricus bisporus are contaminated by Verticillium fungicola. Hydrogen peroxide concentrations were measured in lyophilised samples of bubbles and healthy sporocarps from cultures of genetically related strains of A. bisporus. The strains the more resistant to the pathogen had the higher levels of H2O2 concentration measured in the bubbles, but the differences in the healthy sporocarps were not significant. That is an indication of a higher reaction to the pathogen in the forming sporocarps of A. bisporus strains associated with their partial resistance to V. fungicola.
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    Aspergillus flavus expressed sequence tags for identification of genes with putative roles in aflatoxin contamination of crops (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 237 (2004), S. 0 
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    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Aflatoxins, produced primarily by Aspergillus flavus and A. parasiticus, are among the most toxic and carcinogenic naturally occurring compounds. In an attempt to identify genes potentially involved in aflatoxin contamination of crops, and to better understand the biology of A. flavus, a large scale sequencing of A. flavus expressed sequence tags (EST) was conducted. The 5′ ends of 26,110 cDNA clones from a normalized cDNA expression library were sequenced. After annotation, a total of 7218 unique ESTs in A. flavus were assembled into 3749 tentative concensus sequences and 3469 singleton sequences. The functional classifications of the genes or Gene Ontology (GO) terms were assigned to these ESTs. Genes potentially involved in the aflatoxin contamination process were identified in the ESTs sequenced. These include the aflatoxin biosynthetic pathway, signal transduction, global regulation, pathogenicity of the fungus, and stress response.
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    Nisin-controlled expression of Norwalk virus VP60 protein in Lactobacillus casei (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 237 (2004), S. 0 
    Details
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: A food-grade strain with nisRK stably integrated into the genome, was constructed in order to implement the nisin-controlled expression system (NICE) in Lactobacillus casei ATCC393. Expression of β-glucuronidase (gus) reporter gene was employed to optimize the system, which has been successfully used to produce the main antigenic protein from Norwalk virus, opening new perspectives for producing edible vaccines.
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    Molecular and functional characterization of the Azorhizobium caulinodans ORS571 hydrogenase gene cluster (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 237 (2004), S. 0 
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    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: In this work, we report the cloning and sequencing of the Azorhizobium caulinodans ORS571 hydrogenase gene cluster. Sequence analysis revealed the presence of 20 open reading frames hupTUVhypFhupSLCDFGHJK hypABhupRhypCDEhupE. The physical and genetic organization of A. caulinodans ORS571 hydrogenase system suggests a close relatedness to that of Rhodobacter capsulatus. In contrast to the latter species, a gene homologous to Rhizobium leguminosarum hupE was identified downstream of the hyp operon. A hupSL mutation drastically reduced the high levels of hydrogenase activity induced by the A. caulinodans ORS571 wild-type strain in symbiosis with Sesbania rostrata plants. However, no significant effects on dry weight and nitrogen content of S. rostrata plants inoculated with the hupSL mutant were observed in plant growth experiments.
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    FLO11 is essential for flor formation caused by the C-terminal deletion of NRG1 in Saccharomyces cerevisiae (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 237 (2004), S. 0 
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    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: The flor strains of Saccharomyces cerevisiae form a flor on the surface of wine after alcoholic fermentation. High hydrophobicity of the cell surface is suggested to be important for flor formation by the flor wine yeasts. However, the molecular mechanism of flor formation is not clear. We found that expression of C-terminal deleted NRG1 lacking its two C2H2 zinc finger motifs (NRG11–470) on the multicopy plasmid conferred the ability to form a flor to a non-flor laboratory strain. The cell surface hydrophobicity of NRG11–470 was higher than of the non-flor strain. Disruption of the Nrg1p-repressed gene FLO11, which encodes a cell surface glycoprotein that functions as a flocculin or an adhesin, abolished flor formation. Moreover, expression of FLO11 on a multicopy plasmid could also cause flor formation. These results indicate that FLO11 is essential for flor formation by NRG11–470. In addition, the results suggest that the C-terminal truncated form of Nrg1p exerts a dominant negative effect on FLO11 repression, resulting in FLO11 expression and, thus, flor formation.
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    Regulation of polyhydroxyalkanoate biosynthesis in Pseudomonas putida and Pseudomonas aeruginosa (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 237 (2004), S. 0 
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    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Regulation of medium-chain-length polyhydroxyalkanoate biosynthesis in Pseudomonas aeruginosa and Pseudomonas putida was studied conducting PHA accumulation experiments and transcriptional analysis of PHA biosynthesis genes with wild type strains and rpoN-negative mutants. In P. putida PHA accumulation was RpoN-independent, whereas in P. aeruginosa PHA accumulation was RpoN-dependent. Transcriptional analysis applying reverse transcriptase-polymerase chain reaction showed strong induction of phaG, encoding the transacylase, under nitrogen starvation in P. putida KT2440 and the respective rpoN-negative mutant, indicating an RpoN-independent regulation of phaG. No transcription of phaG and no PHA accumulation was detected in the rpoN-negative mutant of P. aeruginosa neither from gluconate nor from octanoate as carbon source. Alginate-overproducing mutant P. aeruginosa FRD1 showed strongly decreased PHA accumulation from gluconate but no difference in phaC1 (encoding the PHA synthase) transcription, indicating that alginate biosynthesis competes with PHA biosynthesis regarding acetyl-CoA as precursor for both biopolymers. Transcription of phaF and phaI-F was nitrogen independent.
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    Mobile DNA elements in the gas vesicle gene cluster of the planktonic cyanobacteria Microcystis aeruginosa (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 237 (2004), S. 0 
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    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Insertion sequences (IS) have been characterized in Microcystis aeruginosa gas vesicle-deficient mutants. ISMae4, a homolog of the cyanobacterial IS702, belongs to the IS5 family, subgroup ISL2. ISMae2 and ISMae3 display typical IS features and express a transposase of the IS4 and IS1 family, respectively. ISMae1 exhibits a more complex genetic structure and harbours a degenerated transposase gene distantly related to IS1 elements. Hybridizations with IS-specific DNA probes suggest that transposition of ISMae2 and ISMae3 occurred by a replicative-type mechanism. To our knowledge this is the first report showing that IS1 elements can be mobile in cyanobacteria.
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    Isolation and characterization of the Rhodococcus opacus thiostrepton-inducible genes tipAL and tipAS: application for recombinant protein expression in Rhodococcus (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 237 (2004), S. 0 
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    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: We cloned the Rhodococcus opacus (strain DSM 44193) tipA gene, which encodes two translation products, TipAL and TipAS. The gene products are homologous to the Streptomyces spp. TipAL and TipAS proteins, respectively. The tipA promoter is highly active and TipAS protein is predominantly accumulated in R. opacus cells when the inducer of transcription, thiostrepton, was presented in culture medium. We found that thiostrepton is also induced the expression of an endogenous TipA-family protein in Rhodococcus erythropolis (strain JCM3201). The minimal tipA promoter region was defined (57 bp) and the conserved nucleotide sequence of the putative TipAL protein binding site (TipA-box) was identified in that region. The tipA gene is presumed to be transcribed into a leaderless mRNA. We applied the tipA promoter successfully for recombinant protein expression in R. erythropolis cells.
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    Fluorescence measurements of free [Mg2+] by use of mag-fura 2 in Salmonella enterica (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 237 (2004), S. 0 
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    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: The Mg2+ fluorescent dye mag-fura 2, entrapped in cells or organelles, has frequently been used for dual excitation ratio-metric determinations of free ionic Mg2+ concentrations in eukaryotic, mostly mammalian cells. Here we report its successful application to measure free Mg2+ concentrations ([Mg2+]i) in Salmonella enterica cells. When kept in nominally Mg2+ free buffer (resting conditions), the [Mg2+]i of wild-type cells has been determined to be 0.9 mM. An increase in the external Mg2+ concentration ([Mg2+]e) resulted in a rapid increase of [Mg2+]i, saturating within a few seconds at about 1.5 mM with [Mg2+]e of 20 mM. In contrast, cells lacking the Mg2+ transport proteins CorA, MgtA, MgtB failed to show this rapid increase. Instead, their [Mg2+]i increased steadily over extended periods of time and saturated at concentrations below those of wild-type cells. Mg2+ uptake rates increased more than 15-fold when corA was overexpressed in these mutant cells. Uptake of Mg2+ into corA expressing cells was strongly stimulated by nigericin, which increased the membrane potential ΔΨ at the expense of ΔpH, and drastically reduced by valinomycin, which decreased the membrane potential ΔΨ. These results reveal mag-fura 2 as a useful indicator to measure steady-state [Mg2+]i values in resting bacterial cells and to determine Mg2+ uptake rates. They confirm the role of CorA as the major Mg2+ transport protein and reveal the membrane potential as driving force for Mg2+ uptake into S. enterica cells.
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    Extensive set of mitochondrial LSU rDNA-based oligonucleotide probes for the detection of common airborne fungi (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 237 (2004), S. 0 
    Details
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Fungi exist in every indoor and outdoor environment. Many fungi are toxigenic or pathogens that may cause various public health concerns. Rapid and accurate detection and identification of fungi require specific markers. In this study, partial mitochondrial large subunit rDNA was amplified and sequenced from 32 fungal strains representing 31 species from 14 genera. Based on the sequence variation pattern, 26 oligonucleotide probes were designed for their discrimination. The specificity of the probes was evaluated through homology search against GenBank database and hybridization examination on 38 fungal strains. The 26 probes were verified as highly specific to 20 fungal species. A two-step detection procedure through PCR followed by probe hybridization gave ten-fold increase in detection sensitivity than single-step PCR assay and would be a practical approach for environmental sample screening. The probes developed in this study can be applied in clinical diagnosis and environmental monitoring of fungal agents.
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    In vivo induced antigenic determinants of Actinobacillus actinomycetemcomitans (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 237 (2004), S. 0 
    Details
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Actinobacillus actinomycetemcomitans is a Gram-negative capnophilic rod and the etiological agent of localized aggressive periodontitis. The genome-wide survey of A. actinomycetemcomitans using in vivo induced antigen technology (IVIAT) has previously resulted in the discovery of antigenic determinants expressed specifically in diseased patients. The present study evaluated the potential of these antigens as putative disease markers, and investigating their contribution to the pathogenesis of the microorganism. Sera from patients had a significantly greater antibody titer than sera from healthy controls against six antigens, which supports the in vivo expression of these antigens, and suggests their usefulness as disease markers. A. actinomycetemcomitans invasion of epithelium-derived HeLa cells resulted in the induction of all three genes tested, as evidenced by real-time PCR. Isogenic mutants of these three genes were constructed and the adhesion and intracellular survival of the mutants was assayed in a competition assay with the wild-type strain. A significant defect in the intracellular survival of two of these mutant strains (orf1402 and orf859) was found. This defect could not be attributed to an adhesion defect. In contrast, a mutation in vapA, a homologue of a novel putative transcriptional regulator, out-competed the wild-type strain in the same assay. The virulent phenotype was restored for a mutant strain in orf859 upon complementation. This data provided new insight into the pathogenic personality of A. actinomycetemcomitans in vivo and supported the use of HeLa cells as a valid in vitro host–pathogen interactions model for that microorganism. IVIAT is applicable to most pathogens and will undoubtedly lead to the discovery of novel therapies, antibiotics and diagnostic tools.
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    Novel protein domains and motifs in the marine planctomycete Rhodopirellula baltica (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 236 (2004), S. 0 
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    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: The planctomycetes are a phylum of bacteria that have a unique cell compartmentalisation and yeast-like budding cell division and peptidoglycan-less proteinaceous cell walls. We wished to further our understanding of these unique organisms at the molecular level by searching for conserved amino acid sequence motifs and domains in the proteins encoded by Rhodopirellula baltica. Using BLAST and single-linkage clustering, we have discovered several new protein domains and sequence motifs in this planctomycete. R. baltica has multiple members of the newly discovered GEFGR protein family and the ASPIC C-terminal domain family, whilst most other organisms for which whole genome sequence is available have no more than one. Many of the domains and motifs appear to be restricted to the planctomycetes. It is possible that these protein domains and motifs may have been lost or replaced in other phyla, or they may have undergone multiple duplication events in the planctomycete lineage. One of the novel motifs probably represents a novel N-terminal export signal peptide. With their unique cell biology, it may be that the planctomycete cell compartmentalisation plan in particular needs special membrane transport mechanisms. The discovery of these new domains and motifs, many of which are associated with secretion and cell-surface functions, will help to stimulate experimental work and thus enhance further understanding of this fascinating group of organisms.
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    Presence of drug resistance in intestinal lactobacilli of dairy and human origin in Turkey (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 236 (2004), S. 0 
    Details
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: The prevalence of different resistance genes was investigated in lactobacilli of human and dairy origin by PCR. The presence of erm, van, tet, and cat-TC genes were determined in 16 raw milk, 15 cream, 10 yogurt, 50 hand-made cheese, and 20 industrially produced white-cheese samples of dairy origin and 16 mouth, 32 fecal, and 36 vaginal samples from different subjects of human origin. Lactobacilli of dairy and human origin were found to carry only erm(B) and tet(M) genes. The majority of the isolates, Lactobacillus crispatus (61), Lactobacillus gasseri (49), Lactobacillus plantarum (80) studied were found to harbor either erm(B) or tet(M) gene or both. No resistant lactobacilli was found in raw-milk and cream samples. All the human fecal samples and the majority of vaginal (29 of 36) and mouth (10 of 14) samples were found to carry the resistance genes. While a third of the hand-made cheeses carried resistant lactobacilli only one industrially produced cheese was found to carry resistant lactobacilli. Furthermore, the genes were found in the non-starter species, Lactobacillus acidophilus and Lb. plantarum, indicating that industrially produced cheeses in this respect could be considered more favorable. These results indicate that drug resistance seems to be very common in Turkey. Even though the number of dairy samples harboring the resistance genes (17 of 111) is smaller in regards to human samples, 10% of them were still found to carry the resistance genes as well. The presence of the resistance genes in majority of the samples of human origin and in minority of the samples of dairy origin indicates that drug resistance may be acquired in the intestinal tract during passage and spread to dairy products by the hands of workers during production.
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    Sinorhizobium medicae genes whose regulation involves the ActS and/or ActR signal transduction proteins (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 236 (2004), S. 0 
    Details
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: ActS–ActR proteins belong to a highly conserved family of two-component signal transduction systems involved in global regulation in the α-proteobacteria; they were first identified in Sinorhizobium medicae (previously Sinorhizobium meliloti) as essential for acid-tolerance. This paper reports on the identification of genes regulated by ActS and/or ActR in S. medicae. To do this, random gusA fusions were created in S. medicae to follow gene transcription in an actS chromosomal knockout mutant containing plasmid-borne actS. Plasmid borne actS was cured from the mutants and β-glucuronidase (GUS) activity compared between the different genetic backgrounds. We detected actS-dependent regulation of the genes gst1 (detoxification), hyuA (hydantoin utilization) and fixN2 (microaerobic respiration). We show that ActR is involved in regulating cbbS (CO2 fixation), narB (nitrate assimilation) and required for low pH and microaerobic induction of the nitrogen fixation regulators fixK and nifA. In particular, we demonstrate that the transcriptional activation of fixN2 is regulated by ActR through FixK.
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    Characterization of the replicon of a 51-kb native plasmid from the gram-positive bacterium Leifsonia xyli subsp. cynodontis (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 236 (2004), S. 0 
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    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: The 4992-bp replicon of a large cryptic plasmid in the gram-positive bacterium Leifsonia xyli subsp. cynodontis was identified and sequenced. The replicon encoded two proteins essential for plasmid replication and stability. The putative replication protein (RepA) is homologous to that of the plasmids in mycobacterial pLR7 family, while the putative ParA protein immediately downstream of RepA is significantly homologous to the Walker-type ATPase required for partition of plasmid and chromosome of the gram-positive bacteria. These two proteins and other ORFs are clustered with the putative promoters and other regulatory sequences, illustrating an efficient organization of the replicon for this novel plasmid.
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    Transformation of colonial Escherichia coli on solid media (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 236 (2004), S. 0 
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    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: We report that colonial Escherichia coli cells on various solid media can develop modest genetic competence. Using an on-filter culture system, we found that E. coli colonies on CaCl2-containing agar were transformed in the presence of plasmid DNA. Interestingly, transformation also occurred on LB-agar, various moist foods and even on H2O-agar. These results suggest that some populations of colonial E. coli in various environments could become transformable regardless of the surrounding Ca2+ concentration.
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    A new vector, pGID052, for genetic transfer in Oenococcus oeni (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 236 (2004), S. 0 
    Details
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Despite the large number of techniques available for the transformation of bacteria, several species are still resistant to the introduction of foreign DNA. Oenococcus oeni are among the organisms that are particularly refractory to transformation. However, conjugal experiments from Lactococcus lactis to O. oeni with a new plasmid, pGID052, were performed via mobilization with success. This plasmid, a derivative of pORI19, encompasses: (i) the oriT of pIP501, which permitted the transfer to O. oeni, (ii) the replication genes of a native Leuconostoc citreum plasmid. Frequencies of 10−7 conjugants per recipient were found. The transfer did not affect the structure of this low-copy-number plasmid. Moreover, pGID052 seems segregationally stable and could be used in the future as an expression vector.
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    The 3′ termini of small RNAs in Trypanosoma brucei (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 236 (2004), S. 0 
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    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Post-transcriptional 3′ end formation is an essential step in the maturation of most small RNAs. Knowledge of the precise 3′ ends of mature RNAs is essential for defining 3′-processing activities. We have mapped the mature 3′ ends of Spliced Leader RNA, the U1, U2, U4, U5, and U6 small nuclear RNAs, the U3 small nucleolar RNA, and the 5S and 5.8S ribosomal RNAs by ligation-mediated PCR in Trypanosoma brucei. With the exception of U5, two classes of 3′ ends were observed: flush with the base of a stem–loop structure, and 1–2 nt extended from a stem–loop. Multiple mechanisms and structural features are likely to influence 3′ end maturation of RNAs.
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    Methylthioacetaldehyde, a possible intermediate metabolite for the production of volatile sulphur compounds from l-methionine by Lactococcus lactis (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 236 (2004), S. 0 
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    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Volatile sulphur compounds (VSCs) production from l-methionine was studied in Lactococcus lactis. In vitro studies with radiolabelled l-methionine and resting cells of L. lactis revealed that l-methionine was initially converted to α-keto-γ-methylthiobutyrate (KMBA) by a transamination reaction. A part of KMBA was subsequently chemically converted to methylthioacetaldehyde, methanethiol and dimethylsulphides. Chemical conversion of KMBA to methylthioacetaldehyde was dependent on pH, Mn(II) and oxygen. Since methanethiol and dimethylsulphide production was highly related to that of methylthioacetaldehyde, the latter compound was proposed as being an intermediate in VSCs production by L. lactis.
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    Inhibition of Salmonella enterica serovars by microcin J25 (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 236 (2004), S. 0 
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    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Escherichia coli microcin J25 (MccJ25) is a 2107-Da peptide antibiotic whose uptake into E. coli is mediated by the outer-membrane receptor FhuA and the inner membrane proteins TonB, ExbB, ExbD, and SbmA. A survey of the sensitivity of several Salmonella enterica serovars showed that the antibiotic was highly active against some serovars, while S. Typhimurium, S. Derby, and some S. Enteritidis strains were completely resistant. Resistant strains became hypersensitive to MccJ25 when given the fhuA gene of E. coli, indicating that insensitivity is due to the inability of the FhuA protein to mediate penetration of MccJ25. Whereas in E. coli MccJ25 targets RNA polymerase, in S. Typhimurium it inhibits not only RNA synthesis but also cell respiration. Fluorescence viability staining showed that S. Typhimurium cells exposed to MccJ25 remain viable but are unable to form colonies.
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    Promoted proliferation of an SOD-deficient mutant of Escherichia coli under oxidative stress induced by photoexcited TiO2 (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 236 (2004), S. 0 
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    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: An SOD null mutant of Escherichia coli (IM303) and its wild-type strain (MM294) were cultivated with or without sublethal oxidative stress generated from photoexcited TiO2. Concerning maximum specific growth rate of the cells, μm, measured under various conditions, the μm value of IM303 cells increased notably in the presence of TiO2 illuminated with light (I=12.5 W m−2), being about two times higher than that of the cells grown in the absence of TiO2 and light. The μm value of IM303 cells under the oxidative condition restored to a level comparable to that of wild-type MM294 cells, which coincided with the finding that the content of reactive oxygen species lowered in IM303 cells under the oxidative stress. Colony isolation was conducted to obtain the cells prevailing in the early culture phase of IM303 cells in the presence of TiO2 and light. It was found that the isolates exhibited the outgrowing properties with the increased μm values under both the conditions with and without TiO2 and light. It was also indicated that in the culture of typically selected isolate, the cells started to grow with a relatively short lag in a threonine-minus medium.
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    Metalloregulation in Bacillus subtilis: the copZ chromosomal gene is involved in cadmium resistance (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 236 (2004), S. 0 
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    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: The copZ gene of Bacillus subtilis encodes a copper chaperon CopZ that donates copper to the copper transporter CopA. Both genes copZ and copA are clustered to an operon and its promoter is regulated by Cu ions and CueR, a Mer-like transcriptional activator. Here we show that cadmium ions activate copZA expression as strong as copper ions. Northern hybridization analysis showed that copper and cadmium both induce the synthesis of a 2.7 kb copZA transcript and a 250 bp copZ transcript. A copA deletion mutant was sensitive to copper, whereas a copZ deletion resulted in an increased sensitivity to cadmium and copper. Transcription of the cadmium resistance gene cadA, which is adjacent to the copZA cluster, is extremely reduced in a copZ deletion strain. Transformation of copZ in trans restores wild type resistance to cadmium and copper in a copZ deletion strain. This excludes any polar effect and proves that the copZ encoded protein is important for copper and cadmium resistance.
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    Partial purification and biochemical characterization of a soluble α-glucosidase II-like activity from Candida albicans (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 236 (2004), S. 0 
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    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: A soluble α-glucosidase was partially purified from Candida albicans cells by a three-step procedure consisting of size-exclusion, ion-exchange and adsorption chromatographies. After the last step, enzyme was enriched about 8.7-fold with a yield of 13% over the starting material and analysis of the purified preparation revealed two major polypeptides of 36 and 47 kDa. The latter was responsible for enzyme activity as visualized with a fluorescent substrate. Nigerose, an α-1,3-linked glucose disaccharide, was preferentially hydrolyzed by the purified enzyme over other glucosedisaccharides bearing distinct α-linkages. The purified α-glucosidase also converted the GlcMan9GlcNAc2 oligosaccharide into the Man9GlcNAc2 product in a time-dependent manner. These and other determined properties are consistent with a type GII α-glucosidase probably involved in N-glycan processing.
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    Corrigendum to: “An ffh mutant of Streptococcus mutans is viable and able to physiologically adapt to low pH in continuous culture” [FEMS Microbiol. Lett. 234 (2004) 315–324] (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 236 (2004), S. 0 
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    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
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    Developing a quantitative approach for determining the in vitro prebiotic potential of dietary oligosaccharides (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 236 (2004), S. 0 
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    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Prebiotics are nondigestible carbohydrates that beneficially affect the host by selectively stimulating the growth and/or activity of one, or a limited number of, bacteria present in the colon. The selected genera should have the capacity to improve host health (e.g. Bifidobacterium, Lactobacillus). To help identify preferred types, for inclusion into the diet, a quantitative equation [measure of the prebiotic effect (MPE)] is suggested. This will help evaluate, in vitro, the fermentation of dietary carbohydrates and compare their prebiotic effect. Although the approach is not meant to define health values, it is formulated to better inform the choice of prebiotic. It therefore, compares measurements of bacterial changes through the determination of maximum growth rates of predominant groups present in faeces, rate of substrate assimilation and the production of lactic, acetic, propionic and butyric acids. The equation will allow further in vitro comparisons of MPE, leading towards further studies (e.g. in humans) to determine the success of dietary intervention.
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    Macrofilamentous microbial communities in the metal-rich and acidic River Tinto, Spain (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 235 (2004), S. 0 
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    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: A novel type of macroscopic microbial community consisting of large dendritic filaments (up to 1.5 m) in a pH 2.0 dam of the River Tinto (South-western Spain) is described. The combined use of 16S rRNA-gene surveys and fluorescent in situ hybridisation (FISH) suggested that γ-proteobacteria and a relative large diversity of α-proteobacteria dominated these structures. β-Proteobacteria, Actinobacteria and Firmicutes were also detected. Whereas acidophilic bacteria of the genera Acidithiobacillus, Leptospirillum and Acidiphilium, and archaea belonging to the Thermoplasmatales dominate mine acid drainage waters and streamers (riverbed filamentous biofilms), none of the lineages identified in this study affiliate to typical acid mine drainage acidophilic bacteria. Bacteria of the Tinto macrofilaments might be heterotrophic, and could be feeding on the organic matter entrapped in the filamentous structure.
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    The Erp protein is anchored at the surface by a carboxy-terminal hydrophobic domain and is important for cell-wall structure in Mycobacterium smegmatis (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 231 (2004), S. 0 
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    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Erp (exported repetitive protein), also known as P36, Pirg and Rv3810, is a member of a mycobacteria-specific family of extracellular proteins. In pathogenic species, the erp gene has been described as a virulence factor. The Erp proteins comprise three domains. The N- and C-terminal domains are similar in all mycobacterial species, while the central domain consists of a repeated module that differs considerably between species. Here we show that the Erp protein is loosely attached to the surface and that the carboxy-terminal domain, which displays hydrophobic features, anchors Erp at the surface of the bacillus. The hydrophobic region is not necessary for the complementation of the altered colony morphology of a Mycobacterium smegmatis erp− mutant but proved to be necessary to achieve resistance to detergent at wild-type levels.
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    Disruption of response regulator gene, devR, leads to attenuation in virulence of Mycobacterium tuberculosis (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 231 (2004), S. 0 
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    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: The devR-devS two-component system of Mycobacterium tuberculosis was identified earlier and partially characterized in our laboratory. A devR::kan mutant of M. tuberculosis was constructed by allelic exchange. The devR mutant strain showed reduced cell-to-cell adherence in comparison to the parental strain in laboratory culture media. This phenotype was reversed on complementation with a wild-type copy of devR. The devR mutant and parental strains grew at equivalent rates within human monocytes either in the absence or in the presence of lymphocytic cells. The expression of DevR was not modulated upon entry of M. tuberculosis into human monocytes. However, guinea pigs infected with the mutant strain showed a significant decrease in gross lesions in lung, liver and spleen; only mild pathological changes in liver and lung; and a nearly 3 log lower bacterial burden in spleen compared to guinea pigs infected with the parental strain. Our results suggest that DevR is required for virulence in guinea pigs but is not essential for entry, survival and multiplication of M. tuberculosis within human monocytes in vitro. The attenuation in virulence of the devR mutant in guinea pigs together with DevR-DevS being a bona fide signal transduction system indicates that DevR plays a critical and regulatory role in the adaptation and survival of M. tuberculosis within tissues.
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    Survey of the number of two-component response regulator genes in the complete and annotated genome sequences of prokaryotes (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 231 (2004), S. 0 
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    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: The numbers of potential response regulator genes were determined from the complete and annotated genome sequences of Archaea and Bacteria. The numbers of each class of response regulators are shown for each organism, determined principally from BLASTP searches, but with reference to the gene category lists where available. The survey shows that for Bacteria there is a link between the total number of potential response regulator genes and both the genome complexity (number of potential protein-coding genes) and the organism's lifestyle/habitat. Increasingly complex lifestyles and genome complexities are matched by an increase in the average number of potential response regulator genes per genome, indicating that a higher degree of complexity requires a higher level of control of gene expression and cellular activity. Detailed results of this study are available online at and .
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    Disruption of dTDP-rhamnose biosynthesis modifies lipopolysaccharide core, exopolysaccharide production, and root colonization in Azospirillum brasilense (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 231 (2004), S. 0 
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    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: The interaction between Azospirillum brasilense and plants is not fully understood, although several bacterial surface components like exopolysaccharides (EPS), flagella, and capsular polysaccharides are required for attachment and colonization. While in other plant–bacteria associations (Rhizobium–legume, Pseudomonas–potato), lipopolysaccharides (LPS) play a key role in the establishment of an effective association, their role in the root colonization by Azospirillum had not been determined. In this study, we isolated a Tn5 mutant of A. brasilense Cd (EJ1) with an apparently modified LPS core structure, non-mucoid colony morphology, increased EPS production, and affected in maize root colonization. A 3790-bp region revealed the presence of three complete open reading frames designated rmlC, rmlB and rmlD. The beginning of a fourth open reading frame was found and designated rmlA. These genes are organized in a cluster which shows homology to the cluster involved in the synthesis of dTDP-rhamnose in other bacteria. Additionally, the analysis of the monosaccharide composition of LPSs showed a diminution of rhamnose compared to the wild-type strain.
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    Role of benzylsuccinate in the induction of the tutE tutFDGH gene complex of T. aromatica strain T1 (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 231 (2004), S. 0 
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    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Expression of the tutE tutFDGH gene cluster of Thauera aromatica strain T1 was examined by Northern and Western analysis in a wild-type strain and chromosomally deleted strains with or without in-frame deletion plasmids. While expression was observed when the wild-type strain was induced with toluene, various chromosomally deleted strains exhibited little or no expression of the tut genes. In contrast, both wild-type and chromosomally deleted strains expressed the tut genes when induced with benzylsuccinate. We conclude that benzylsuccinate is required for the full induction of the tutE tutFDGH gene cluster of T. aromatica strain T1.
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    Environmental genomics, the big picture? (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 231 (2004), S. 0 
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    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: The enormous sequencing capabilities of our times might be reaching the point of overflowing the possibilities to analyse data and allow for a feedback on where to focus the available resources. We have now a foreseeable future in which most bacterial species will have an annotated genome. However, we know also that most prokaryotic diversity would not be included there. On the one hand, there is the problem of many groups not being easily amenable to culture and hence not represented in culture-centred microbial taxonomy. On the other hand, the gene pools present in one species can be orders of magnitude larger that the genome of one strain (selected for genome sequencing). Contrasting with eukaryotic genomes, the repertoire of genes present in one prokaryotic cell genome does not correlate stringently with its taxonomic identity. Hence gene catalogues from one environment might provide more meaningful information than the classical species catalogues. Metagenomics or microbial environmental genomics provide a different tool that gravitates around the habitat rather than the species. Such tool could be just the right way to complement ‘organismal genomics’. Its potential to advance our understanding of microbial ecology and prokaryotic diversity and evolution is discussed.
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    Mycobacterium smegmatis displays the Mycobacterium tuberculosis virulence-related neutral red character when expressing the Rv0577 gene (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 231 (2004), S. 0 
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    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Neutral red staining is a cytochemical reaction that has been found to be related to Mycobacterium tuberculosis virulence and, therefore, the component involved in it is thought to be a virulence factor. To study the molecular basis of this reaction we constructed an M. tuberculosis cosmid library in Mycobacterium smegmatis and selected recombinant neutral red positive clones. Heterologous complementation identified Rv0577 as the gene responsible for this trait and we have also shown that it is expressed as a single polycistronic unit together with Rv0576 which could also be involved in the neutral red staining.
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    Insertion of an E. coli lacZ gene in Acetobacter xylinus for the production of cellulose in whey (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 231 (2004), S. 0 
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    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: A mini-Tn10:lacZ:kan was inserted into a wild-type strain of Acetobacter xylinus by random transposon mutagenesis, generating a lactose-utilising and cellulose-producing mutant strain designated ITz3. Antibiotic selection plate assays and Southern hybridisation revealed that the lacZ gene was inserted once into the chromosome of strain ITz3 and was stably maintained in non-selective medium after more than 60 generations. The modified strain had, on the average, a 28-fold increase in cellulose production and a 160-fold increase in β-galactosidase activity when grown in lactose medium. β-Galactosidase activity is present in either lactose or sucrose medium indicating that the gene is constitutively expressed. Cellulose and β-galactosidase production by the modified strain was also evaluated in pure and enriched whey substrates. Utilisation of lactose in whey substrate by ITz3 reached 17 g l−1 after 4 days incubation.
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    Chemotaxis proteins and transducers for aerotaxis in Pseudomonas aeruginosa (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 231 (2004), S. 0 
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    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: It was previously shown that the chemotaxis gene cluster 1 (cheYZABW) was required for chemotaxis. In this study, the involvement of the same cluster in aerotaxis is described and two transducer genes for aerotaxis are identified. Aerotaxis assays of a number of deletion–insertion mutants of Pseudomonas aeruginosa PAO1 revealed that the chemotaxis gene cluster 1 and cheR are required for aerotaxis. Mutant strains which contained deletions in the methyl-accepting chemotaxis protein-like genes tlpC and tlpG showed decreased aerotaxis. A double mutant deficient in tlpC and tlpG was negative for aerotaxis. TlpC has 45% amino acid identity with the Escherichia coli aerotactic transducer Aer. The TlpG protein has a predicted C-terminal segment with 89% identity to the highly conserved domain of the E. coli serine chemoreceptor Tsr. A hydropathy plot of TlpG indicated that hydrophobic membrane-spanning regions are missing in TlpG. A PAS motif was found in the N-terminal domains of TlpC and TlpG. On this basis, the tlpC and tlpG genes were renamed aer and aer-2, respectively. No significant homology other than the PAS motif was detected in the N-terminal domains between Aer and Aer-2.
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    Regulation of lantibiotic lacticin 481 production at the transcriptional level by acid pH (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 231 (2004), S. 0 
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    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: The lantibiotic lacticin 481 operon (lctAMTFEG) is mainly transcribed from P1 and P3, two promoters lying upstream of lctA. A weak additional promoter allows independent expression of the immunity genes (lctFEG). Lacticin 481 production by Lactococcus lactis is stimulated by the acidification due to lactic acid production, and by artificially lowering the pH of the medium. This regulation occurs at the transcriptional level, since P1 and P3 are both acid-induced. P1 is weaker but more tightly regulated than P3. As no specific regulator is encoded by the lacticin 481 operon, P1 and P3 are likely controlled by a general regulator.
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    The impact of feed composition on biodegradation of benzoate under cyclic (aerobic/anoxic) conditions (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 231 (2004), S. 0 
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    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: The response of a mixed microbial culture to different feed compositions, that is, containing benzoate and pyruvate as sole carbon sources at different levels, was studied in a chemostat with a 48-h hydraulic residence time under cyclic aerobic and anoxic (denitrifying) conditions. The cyclic bacterial culture was well adapted to different feed compositions as evidenced by the lack of accumulation of benzoate or pyruvate in the chemostat. Both the benzoate-degrading capabilities and the in vitro catechol 2,3-dioxygenase (C23DO) activities of the cyclic bacterial cultures were in direct proportion to the flux through the chemostat of the substrate degraded by the pathway containing C23DO, with some exceptions. The quantity of C23DO showed a transient decrease during the initial portion of the aerobic period before returning to the level present during the anoxic period. That decrease was most likely caused by the production of H2O2 by the cells upon being returned to aerobic conditions.
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    MprF-mediated biosynthesis of lysylphosphatidylglycerol, an important determinant in staphylococcal defensin resistance (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 231 (2004), S. 0 
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    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Frequently bacteria are exposed to membrane-damaging cationic antimicrobial molecules (CAMs) produced by the host's immune system (defensins, cathelicidins) or by competing microorganisms (bacteriocins). Staphylococcus aureus achieves CAM resistance by modifying anionic phosphatidylglycerol with positively charged l-lysine, resulting in repulsion of the peptides. Inactivation of the novel S. aureus gene, mprF, which is found in many bacterial pathogens, has resulted in the loss of lysylphosphatidylglycerol (L-PG), increased inactivation by CAM-containing neutrophils, and attenuated virulence. We demonstrate here that expression of mprF is sufficient to confer L-PG production in Escherichia coli, which indicates that MprF represents the L-PG synthase. L-PG biosynthesis was studied in vitro and found to be dependent on phosphatidylglycerol and lysyl-tRNA, two putative substrate molecules. Further addition of cadaverin, a competitive inhibitor of the lysyl-tRNA synthetases, or of RNase A abolished L-PG biosynthesis, thereby confirming the involvement of lysyl-tRNA. This study forms the basis for further detailed analyses of L-PG biosynthesis and its role in bacterial infections.
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    Effect of pH on the activity of bovicin HC5, a bacteriocin from Streptococcus bovis HC5 (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 231 (2004), S. 0 
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    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: The bacteriocin, bovicin HC5, catalyzed potassium efflux from Streptococcus bovis JB1, and this activity was highly pH dependent. When the pH was near neutral, glucose-energized cells were not affected by bovicin HC5, but the intracellular steady-state concentration of potassium decreased at acidic pH values. The idea that pH was affecting bovicin HC5 binding was supported by the observation that acidic pH also enhanced the efflux of potassium from non-energized cells that had been loaded with potassium. The relationship between bovicin HC5 concentration and potassium depletion was a saturation function, but cooperativity plots indicated that the binding of one bovicin molecule to the cell membrane facilitated the binding of another.
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    The Mig protein of Streptococcus dysgalactiae inhibits bacterial internalization into bovine mammary gland epithelial cells (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 231 (2004), S. 0 
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    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: The role of the Mig protein of Streptococcus dysgalactiae in bacterial adhesion and internalization of bovine mammary gland epithelial cells (MAC-T) was investigated with the wild-type and isogenic mig mutant strains. While there was no difference in adhesion between the strains, the wild-type strain exhibited a significantly lower level of invasion than the mutants. The lower level of internalization of the Mig+ strain is likely due to Mig-mediated interference with uptake of the microorganisms rather than the host protein binding properties of Mig. Avoidance of intimate interactions with the host cells might be an alternative strategy for S. dysgalactiae to survive and persist in the bovine mammary glands.
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    Oxalate metabolism by the acetogenic bacterium Moorella thermoacetica (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 231 (2004), S. 0 
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    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Whole-cell and cell-extract experiments were performed to study the mechanism of oxalate metabolism in the acetogenic bacterium Moorella thermoacetica. In short-term, whole-cell assays, oxalate consumption was low unless cell suspensions were supplemented with CO2, KNO3, or Na2S2O3. Cell extracts catalyzed the oxalate-dependent reduction of benzyl viologen. Oxalate consumption occurred concomitant to benzyl viologen reduction; when benzyl viologen was omitted, oxalate was not appreciably consumed. Based on benzyl viologen reduction, specific activities of extracts averaged 0.6 μmol oxalate oxidized min−1 mg protein−1. Extracts also catalyzed the formate-dependent reduction of NADP+; however, oxalate-dependent reduction of NADP+ was negligible. Oxalate- or formate-dependent reduction of NAD+ was not observed. Addition of coenzyme A (CoA), acetyl-CoA, or succinyl-CoA to the assay had a minimal effect on the oxalate-dependent reduction of benzyl viologen. These results suggest that oxalate metabolism by M. thermoacetica requires a utilizable electron acceptor and that CoA-level intermediates are not involved.
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    Survey of molecular methods for the typing of wine yeast strains (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 231 (2004), S. 0 
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    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: A survey of the genetic polymorphisms produced by distinct methods was performed in 23 commercial winery yeast strains. Microsatellite typing, using six different loci, an optimized interdelta sequence analysis and restriction fragment length polymorphism of mitochondrial DNA generated by the enzyme HinfI had the same discriminatory power: among the 23 commercial yeast strains, 21 distinct patterns were obtained. Karyotype analysis gave 22 patterns, thereby allowing the discrimination of one of the three strains that were not distinguished by the other methods. Due to the equivalence of the results obtained in this survey, any of the methods can be applied at the industrial scale.
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    Quinate oxidation in Gluconobacter oxydans IFO3244: purification and characterization of quinoprotein quinate dehydrogenase (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 241 (2004), S. 0 
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    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Quinoprotein quinate dehydrogenase (QDH) is a membrane-bound enzyme containing pyrroloquinoline quinone (PQQ) as the prosthetic group. QDH in Gluconobacter oxydans IFO3244 was found to be inducible by quinate and it is not constitutively expressed in the absence of quinate. The purification of holo-form of QDH to nearly homogeneity was achieved. The purified QDH appears to have two subunits of approximately 65 and 21 kDa, which could be the result of proteolysis of single polypeptide. Kinetic analysis indicated that the purified enzyme is much more specific to quinate than QDH from Acinetobacter calcoaceticus. The efficiency of the artificial electron acceptor was also determined.
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    Analysis of gyrA and parC mutations in enterococci from environmental samples with reduced susceptibility to ciprofloxacin (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 231 (2004), S. 0 
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    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: The quinolone resistance determining regions of gyrA and parC in four species of enterococci from environmental samples with reduced susceptibility to ciprofloxacin were sequenced. The nucleotide sequence variations of parC could be related to the different enterococcal species. Mutations in Enterococcus faecalis and Enterococcus faecium related to reduced susceptibility were identical to mutations detected in E. faecalis and E. faecium of clinical origin. A minimal inhibitory concentration of 8 μg ml−1 to ciprofloxacin was not associated with any mutations in the gyrA and parC gene of Enterococcus casseliflavus and Enterococcus gallinarum. These two species may be intrinsically less susceptible to ciprofloxacin.
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    Two Bacillus sphaericus binary toxins share the midgut receptor binding site: implications for resistance of Culex pipiens complex (Diptera: Culicidae) larvae (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 241 (2004), S. 0 
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    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: This work demonstrates that Bin1 and Bin2 toxins, produced by Bacillus sphaericus strains IAB59 and 2362, respectively, share a binding site in midgut brush border membranes (BBMF) from Culex pipiens complex larvae. However, a colony selected with strain IAB59, displaying a resistance ratio of only 42-fold to IAB59, but a 162,000-fold resistance to strain 2362, was found to miss receptors for Bin2 in the BBMF. This correlates with results showing that Bin1, produced in strain IAB59, failed to bind specifically to BBMF from other colony highly resistant to strain 2362. Data indicate the loss of the BBMF bound receptor as a general mechanism of resistance to binary toxins in mosquito.
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    The Saccharomyces cerevisiae gene ECM11 is a positive effector of meiosis (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 241 (2004), S. 0 
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    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Ecm11 is classified as a protein involved in yeast cell wall biogenesis and organization, but in this paper, we provide evidence that it is involved in meiosis as well. Mutants with deleted ECM11 exhibit complex defects in meiosis: replication, recombination and chromosome segregation are affected. The ecm11Δ diploid strains sporulate more slowly and less efficiently than parental strains with wild type copies of ECM11. Fluorescence activated cell sorter scans of DNA content during sporulation showed that meiotic DNA synthesis is initiated at the same time in parental and ecm11Δ strains, but is less efficient in the knockout strain. By recombination tests, we demonstrated that ECM11 is required for crossing-over, but not for gene conversion. In the absence of ECM11 gene product, viability of spores is reduced to 50% and predominantly two viable spores per tetrad are formed. Our results suggest that ECM11 is required in early stages of meiosis where its function is related to DNA replication and crossing-over.
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    Cytochrome c maturation proteins are critical for in vivo growth of Legionella pneumophila (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 241 (2004), S. 0 
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    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Legionella pneumophila, an intracellular parasite of macrophages and protozoa, requires iron for extra- and intracellular growth. In a new screen of a mutant library of L. pneumophila for strains defective for growth on agar media lacking supplemental iron, seven mutants were obtained. All of the mutants had a disruption in the cytochrome c maturation (ccm) locus; two had insertions in ccmB, two in ccmC, and three in ccmF. The ccm mutants were unable to multiply within macrophage-like cells (i.e., U937 and THP-1 cells) and Hartmannella vermiformis amoebae. A competition assay in A/J mice revealed that ccm mutants are severely defective for growth within the lung. Taken together, these data confirm that ccm and cytochrome c maturation proteins are required for L. pneumophila growth in low iron, intracellular infection, and virulence.
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    Cu/Zn superoxide dismutase in yeast mitochondria – a general phenomenon (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 230 (2004), S. 0 
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    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Fermentative and respiratory yeast strains of genera Saccharomyces, Kluyveromyces, Pichia, Candida and Hansenula have been investigated for mitochondrial localization of Cu/Zn superoxide dismutase (SOD). Pure mitochondrial fractions were obtained and the specific activities of Cu/Zn and Mn SODs were measured in comparison with those in the corresponding cell-free extracts. The Cu/Zn SOD: Mn SOD ratio in mitochondria and crude extracts was calculated and was considered a specific characteristic of all tested strains. Electrophoretical visualization of SOD patterns provided evidence for possible migration of cytosolic Cu/Zn SOD to mitochondria. The characteristic Cu/Zn SOD profile in mitochondria of all tested strains suggested its ubiquity within the fermentative and respiratory yeasts.
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    Biodiversity of cultivable psychrotrophic marine bacteria isolated from Terra Nova Bay (Ross Sea, Antarctica) (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 230 (2004), S. 0 
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    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: A set of 146 Antarctic marine isolates from the Ross Sea was characterized by a combination of molecular techniques in order to determine the degree of inter- and intraspecific variability. Isolates were analyzed by amplified rDNA restriction analysis (ARDRA) using the tetrameric enzyme AluI, resulting in 52 different groups, corresponding to at least 52 different bacterial species, indicating a high degree of interspecific variability. The phylogenetic position of bacteria belonging to some ARDRA groups was obtained by sequencing of 16S rDNA. Random amplified polymorphic DNA (RAPD) analysis, carried out on the largest ARDRA groups, revealed a high intraspecific genetic variability, too. The analysis of plasmid content revealed the existence of horizontal gene transfer between strains belonging to the same and to different species. A comparison of the whole body of morphological, physiological and biochemical data was finally carried out.
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    Cloning, sequencing and functional analysis of Magnaporthe grisea MVP1 gene, a hex-1 homolog encoding a putative ‘woronin body’ protein (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 230 (2004), S. 0 
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    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: A hex-1 homolog named MVP1 was isolated from an appressoria cDNA library of the rice blast fungus Magnaporthe grisea. The transcript of ∼1.6 kb contains 546 bp of coding sequence with a 3′ untranslated region about 168 bp long and a 5′ untranslated region about 870 bp long. Southern gel blot analysis of genomic DNA following digestion with three restriction enzymes (BamHI, EcoRI, HindIII) indicated that the gene exists as a single copy in M. grisea genome. RNA gel blot analyses showed that MVP1 was highly expressed in germinating conidia and the mycelial stage compared to appressoria or non-germinated conidia. MVP1 showed a high degree of homology to the hex-1 gene recently described to encode a major protein in the woronin bodies of Neurospora crassa. Double homologous recombination was used to replace MVP1 with the hygR gene. MVP1 knockout mutants showed apical swellings when grown on agar plates containing 2% sorbose but they were not impaired in any other vegetative or pathogenic properties evaluated. The pathological and other phenotypic consequences of gene disruption are discussed.
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    Expression of the landomycin biosynthetic gene cluster in a PKS mutant of Streptomyces fradiae is dependent on the coexpression of a putative transcriptional activator gene (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 230 (2004), S. 0 
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    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: The formation of landomycin A or one of its derivatives (5,6-anhydrolandomycin A) in a heterologous strain has never been achieved. It has now been made possible by the coexpression of a cosmid containing all biosynthetic genes necessary to produce landomycin A together with a pathway-specific regulatory gene. As host we used a polyketide synthase-defective mutant strain of Streptomyces fradiae Tü2717 which is not able to produce urdamycin A. Our results indicate that four glycosyltransferases are responsible for the formation of the hexasaccharide side chain of landomycin A.
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    URL: http://dx.doi.org/10.1016/S0378-1097(03)00861-9
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    Porphyromonas gingivalis induces its uptake by human macrophages and promotes foam cell formation in vitro (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 241 (2004), S. 0 
    Details
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Porphyromonas gingivalis is an etiologic agent of periodontal disease in humans, which has been linked to an increased risk for atherosclerosis-related events. In this study, we examined the effect of P. gingivalis infection on human macrophages with respect to foam cell formation, the hallmark of early atherogenesis, and the potential of P. gingivalis to induce its uptake by these cells. Human monocyte-derived macrophages were incubated with low density lipoprotein and infected with P. gingivalis FDC381 or its fimbriae deficient mutant, DPG3. Consistent with a role for fimbriae in this process, strain 381 significantly increased foam cell formation as compared to DPG3. Recovery of viable P. gingivalis in antibiotic protection experiments was significantly higher for strain 381 than for DPG3. By transmission electron microscopy, the wild-type strain was shown to adhere to and enter THP-1 cells. These results suggest that properties of P. gingivalis which render it capable of adhering to/invading other cell types may also be operative in macrophages and play an important role in its atherogenic potential.
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    URL: http://dx.doi.org/10.1016/j.femsle.2004.10.009
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    Heterologous production and characterization of bacterial nickel/cobalt permeases (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 230 (2004), S. 0 
    Details
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Nickel/cobalt permeases (NiCoTs, TC 2.A.52) are a rapidly growing family of structurally related membrane transporters whose members are found in Gram-negative and Gram-positive bacteria, in thermoacidophilic archaea, and in fungi. Previous studies have predicted two subclasses represented by HoxN of Ralstonia eutropha, a selective nickel transporter, and by NhlF of Rhodococcus rhodochrous, a nickel and cobalt transporter that displays a preference for the Co ion. In the present study, NiCoT genes of five Gram-negative bacteria and one Gram-positive bacterium were cloned and heterologously expressed in Escherichia coli. Based on substrate preference in metal-accumulation assays with the recombinant strains, two of the novel NiCoTs were assigned to the NhlF class. The remaining four NiCoTs belong to a yet unrecognized, third class. They transport both the nickel and the cobalt ion but have a significantly higher capacity for nickel. The observed substrate preferences correlate in many cases with the genomic localization of NiCoT genes adjacent to regions encoding nickel- or cobalt-dependent enzymes or enzymes involved in cobalamin biosynthesis. Alignment of 23 full-length NiCoT sequences and comparison with the available experimental data predict that substrate specificity of NiCoTs is an adaptation to specific transition metal requirements in various organisms from different taxa.
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    URL: http://dx.doi.org/10.1016/S0378-1097(03)00885-1
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    Notes to authors (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 230 (2004), S. 0 
    Details
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1016/S0378-1097(03)00947-9
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    Cloning and characterization of preferentially expressed genes in an aluminum-tolerant mutant derived from Penicillium chrysogenum IFO4626 (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 230 (2004), S. 0 
    Details
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: cDNAs expressed preferentially in an Al-tolerant microorganism were isolated by subtraction hybridization with cDNAs of Al-sensitive Penicillium chrysogenum IFO4626 as driver cDNA and cDNAs of the Al-tolerant mutant derived from the wild cells by UV irradiation as tester cDNA. Northern blot analysis revealed that mRNA levels of six genes were increased significantly in the Al-tolerant mutant after exposure to Al stress when compared with the wild cells. Two genes accumulated in both the presence and absence of Al stress and four genes were induced by Al stress in the Al-tolerant mutant. cDNA fragments were amplified by rapid amplification of cDNA ends and sequenced to obtain full-length cDNAs of the six genes. Two genes were novel or predicted ones and the others showed significant homology to known genes, ADP/ATP translocase, enolase, cysteine synthase, and glucoamylase, which are induced by environmental stresses in prokaryotic and eukaryotic cells. These enzyme activities increased in the Al-tolerant mutant when compared to those in the wild cells, showing that not only the levels of gene expression but also the levels of enzyme activities increased in the Al-tolerant mutant.
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    URL: http://dx.doi.org/10.1016/S0378-1097(03)00886-3
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  • 89
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    Lifestyle alternatives for rhizobia: mutualism, parasitism, and forgoing symbiosis (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 237 (2004), S. 0 
    Details
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Strains of rhizobia within a single species can have three different genetically determined strategies. Mutualistic rhizobia provide their legume hosts with nitrogen. Parasitic rhizobia infect legumes, but fix little or no nitrogen. Nonsymbiotic strains are unable to infect legumes at all. Why have rhizobium strains with one of these three strategies not displaced the others? A symbiotic (mutualistic or parasitic) rhizobium that succeeds in founding a nodule may produce many millions of descendants. The chances of success can be so low, however, that nonsymbiotic rhizobia can have greater reproductive success. Legume sanctions against nodules that fix little or no nitrogen favor more mutualistic strains, but parasitic strains that use plant resources only for their own reproduction may do well when they share nodules with mutualistic strains.
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    URL: http://dx.doi.org/10.1111/j.1574-6968.2004.tb09695.x
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  • 90
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    Microdiversity of phenol hydroxylase genes among phenol-degrading isolates of Alcaligenes sp. from an activated sludge system (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 237 (2004), S. 0 
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    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Enterobacterial repetitive intergenic consensus (ERIC)-PCR fingerprinting classified 97 phenol-degrading isolates with identical amplified ribosomal DNA restriction analysis (ARDRA) patterns into six genotypic groups. The 16S rRNA gene of the representative isolate of each group had higher than 99.47% common identity with each other and higher than 98% identity with the type strain of Alcaligenes faecalis. PCR-TGGE (temperature gradient gel electrophoresis) analysis of the genes of the largest subunit of the multi-component phenol hydroxylase (LmPH) in each isolate followed with sequencing showed that isolates within each ERIC-PCR group had identical LmPH gene sequences. Among the six different ERIC-PCR groups, two were found to harbor two different LmPH genes encoding low- and high-Ks (affinity constants) phenol hydroxylases, and the low-Ks type LmPH was identical in sequence with one predominant LmPH of the parental activated sludge. Three ERIC-PCR groups had only the high-Ks type and one had no sequence similar to the known LmPHs. Our work suggests that there is no correlation between the phylogenetic groupings of phenol-degrading bacteria and their LmPH genotypes possibly due to extensive horizontal gene transfer of this functional gene.
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    URL: http://dx.doi.org/10.1111/j.1574-6968.2004.tb09719.x
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    Lactoferricin B inhibits bacterial macromolecular synthesis in Escherichia coli and Bacillus subtilis (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 237 (2004), S. 0 
    Details
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Most antimicrobial peptides have an amphipathic, cationic structure, and an effect on the cytoplasmic membrane of susceptible bacteria has been postulated as the main mode of action. Other mechanisms have been reported, including inhibition of cellular functions by binding to DNA, RNA and proteins, and the inhibition of DNA and/or protein synthesis. Lactoferricin B (Lfcin B), a cationic peptide derived from bovine lactoferrin, exerts slow inhibitory and bactericidal activity and does not lyse susceptible bacteria, indicating a possible intracellular target. In the present study incorporation of radioactive precursors into DNA, RNA and proteins was used to demonstrate effects of Lfcin B on macromolecular synthesis in bacteria. In Escherichia coli UC 6782, Lfcin B induces an initial increase in protein and RNA synthesis and a decrease in DNA synthesis. After 10 min, the DNA-synthesis increases while protein and RNA-synthesis decreases significantly. In Bacillus subtilis, however, all synthesis of macromolecules is inhibited for at least 20 min. After 20 min RNA-synthesis increases. The results presented here show that Lfcin B at concentrations not sufficient to kill bacterial cells inhibits incorporation of radioactive precursors into macromolecules in both Gram-positive and Gram-negative bacteria.
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    URL: http://dx.doi.org/10.1111/j.1574-6968.2004.tb09720.x
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    Metabolism of octahydro-1,3,5,7-tetranitro-1,3,5,7-tetrazocine by Clostridium bifermentans strain HAW-1 and several other H2-producing fermentative anaerobic bacteria (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 237 (2004), S. 0 
    Details
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Several H2-producing fermentative anaerobic bacteria including Clostridium, Klebsiella and Fusobacteria degraded octahydro-1,3,5,7-tetranitro-1,3,5,7-tetrazocine (HMX) (36 μM) to formaldehyde (HCHO) and nitrous oxide (N2O) with rates ranging from 5 to 190 nmol h−1 g [dry weight] of cells−1. Among these strains, C. bifermentans strain HAW-1 grew and transformed HMX rapidly with the detection of the two key intermediates the mononitroso product and methylenedinitramine. Its cellular extract alone did not seem to degrade HMX appreciably, but degraded much faster in the presence of H2, NADH or NADPH. The disappearance of HMX was concurrent with the release of nitrite without the formation of the nitroso derivative(s). Results suggest that two types of enzymes were involved in HMX metabolism: one for denitration and the second for reduction to the nitroso derivative(s).
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    URL: http://dx.doi.org/10.1111/j.1574-6968.2004.tb09679.x
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    The rapid identification of European Armillaria species from soil samples by nested PCR (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 237 (2004), S. 0 
    Details
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: New specific primers AR1 and AR2 were successfully used for the amplification of a specific part of internal transcribed spacer (ITS) of rDNA of Armillaria isolated from soil samples. DNA was isolated from 0.5 g of forest soil and ITS region was amplified by nested PCR reaction with external primers ITS1 and ITS4 and internal primers AR1 and AR2. The individual species were distinguished by restriction fragment length polymorphisms (RFLPs) analysis with restriction endonuclease HinfI. The fragments were analysed by ion-exchange HPLC that is more sensible and more rapid than electrophoresis. The amplicons were sequenced to improve the discrimination between the species. The method enables the identification of Armillaria species within one day directly from soil samples without the need for previous isolation and cultivation of mycelium of Armillaria.
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    URL: http://dx.doi.org/10.1111/j.1574-6968.2004.tb09684.x
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    A hydrogen-evolving enzyme is present in Frankia sp. R43 (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 236 (2004), S. 0 
    Details
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: The ability to evolve hydrogen using methyl viologen as an electron donor was assayed in the nitrogen-fixing actinomycetes Frankia sp. R43 and Frankia sp. KB5. To further examine the nature of hydrogen-evolving enzymes that may be present in these organisms immunological studies were performed. Under anaerobic conditions (both nitrogen-limiting and nitrogen-containing) Frankia sp. R43 but not Frankia sp. KB5 evolved hydrogen, which was not linked to NAD-reducing activity. Immunological analysis of total protein from Frankia sp. R43 and Frankia sp. KB5 using an antiserum raised against Ralstonia eutropha HoxF, recognized an antigen in Frankia sp. R43 but not in Frankia sp. KB5. Immunogold labeling using antibodies raised against the R. eutropha HoxH recognized sites in both hyphae and vesicles of Frankia sp. R43, but not in Frankia sp. KB5. Based on these physiological and immunological findings, we conclude that Frankia sp. R43 has a hydrogen-evolving hydrogenase.
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    URL: http://dx.doi.org/10.1111/j.1574-6968.2004.tb09652.x
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    Development of PCR assay to identify Pseudomonas fluorescens and its biotype (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 236 (2004), S. 0 
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    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: The paper provides a simple protocol that uses the polymerase chain reaction to amplify a specific portion of the 16S gene, allowing the recognition of Pseudomonas fluorescens from other group I Pseudomonas. The amplified DNA patterns of 16S rRNA and ITS1, from the restriction fragment length polymorphisms VspI, HaeIII and TaqI digestion, produced band patterns that distinguished the biotypes of Ps. fluorescens. In addition to distinguishing the biotypes C and 3 we used a phenotypical method for levan production.
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    URL: http://dx.doi.org/10.1111/j.1574-6968.2004.tb09655.x
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    Sustained glycolytic oscillations – no need for cyanide (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 236 (2004), S. 0 
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    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Using fluorescence spectroscopy we detected long trains of macroscopic oscillations in the glycolytic pathway, in whole cell suspensions of Saccharomyces cerevisiae, without addition of cyanide. Such oscillations may be induced if argon or another inert gas is bubbled through the yeast cell suspension. This supports that the synchronizing agent is a volatile compound secreted by the yeast cells, e.g. CO2 and/or acetaldehyde. Our results show that the rate of acetaldehyde removal is not a crucial parameter to the synchronization of the yeast cells. The sample cell was connected to a membrane inlet mass spectrometer (MIMS) for online determination of extracellular non-polar compounds. Oscillations in the secretion of CO2 were detected using the MIMS.
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    URL: http://dx.doi.org/10.1111/j.1574-6968.2004.tb09656.x
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    Gene cloning, expression and characterization of novel phytase from Obesumbacterium proteus (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 236 (2004), S. 0 
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    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: The gene phyA encoding phytase was isolated from Obesumbacterium proteus genomic library and sequenced. The cleavage site of the PhyA signal peptide was predicted and experimentally proved. The PhyA protein shows maximum identity of 53% and 47% to phosphoanhydride phosphorylase from Yersinia pestis and phytase AppA from Escherichia coli, respectively. Based on protein sequence similarity of PhyA and its homologs, the phytases form a novel subclass of the histidine acid phosphatase family. To characterize properties of the PhyA protein, we expressed the phyA gene in E. coli. The specific activity of the purified recombinant PhyA was 310 U mg−1 of protein. Recombinant PhyA showed activity at pH values from 1.5 through 6.5 with the optimum at 4.9. The temperature optimum was 40–45 °C at pH 4.9. The Km value for sodium phytate was 0.34 mM with a Vmax of 435 U mg−1.
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    URL: http://dx.doi.org/10.1111/j.1574-6968.2004.tb09659.x
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    Over-production of dihydrofolate reductase leads to sulfa-dihydropteroate resistance in yeast (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 236 (2004), S. 0 
    Details
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Dihydropteroate synthase (DHPS) can metabolise sulfa drugs into sulfa-dihydropteroate (sulfa-DHP), which inhibits cell growth through competition with dihydrofolate (DHF), possibly indicating dihydrofolate reductase (DHFR) as the target of sulfa-DHP. The effect of over-production of DHFR on sulfa-DHP resistance was examined in Saccharomyces cerevisiae using a strain that requires DHF for growth. This strain was transformed with a plasmid which encodes over-production of DHFR in the presence of CuSO4. Over-production led to resistance to sulfa-DHP suggesting that sulfa-DHP targets DHFR. Spontaneous mutants hyper-resistant to sulfa-DHP did not show any changes within DHFR.
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    URL: http://dx.doi.org/10.1111/j.1574-6968.2004.tb09661.x
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    Cloning, expression, and fibrin (ogen)olytic properties of a subtilisin DJ-4 gene from Bacillus sp. DJ-4 (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 236 (2004), S. 0 
    Details
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Previously, we purified a strong fibrinolytic enzyme (subtilisin DJ-4) from Bacillus sp. DJ-4 and characterized its enzymatic activity. Here, we cloned the gene subtilisin DJ-4, and determines its nucleotide sequence, which showed 97% identity with subtilisin BPN′ from B. amyloliquefacens. Recombinant full-subtilisin DJ-4 (rf-subDJ-4) and mature-subtilisin DJ-4 (rm-subDJ-4) were expressed using a pET29 vector system, and their fibrin (ogen)olytic and plasminogen activator activities were studied. rf-subDJ-4 was found to have a higher stability to heat (60 °C) and to acidic conditions (pH 3.0–4.0) than the native subtilisin DJ-4 of Bacillus sp. DJ-4. The plasminogen activator activity of rf-subDJ-4 was 2.75 times greater than that of plasmin on a molar basis. And its specific activity (F/C, the ratio of fibrinolytic activity to caseinolytic activity) was 2.67 and 3.97 times higher than those of subtilisin BPN′ and subtilisin Carlsberg, respectively. rf-subDJ-4 rapidly hydrolyzed the Aα-, Bβ-, and γ-chains of fibrinogen within 5 min. But, unlike subtilisin BPN′ at a very low concentration (50 ng), the γ-chain was not cleaved. On the other hand, rm-subDJ-4 did not show enzyme activity.
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    URL: http://dx.doi.org/10.1111/j.1574-6968.2004.tb09665.x
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    Mechanisms of G protein-coupled receptor-mediated degranulation (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 236 (2004), S. 0 
    Details
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Mast cells and neutrophils play a major role in the innate immune response. Following invasion of the host by microorganisms, these immune cells become activated and release anti-microbial cytotoxic granules in an effort to destroy invading microorganisms in a process termed degranulation. By-products from the degradation of microorganisms can also activate G-protein-coupled receptors (GPCRs), which can further activate immune cells. While degranulation of basophils has been extensively characterized for IgE receptors, the signaling pathways initiated by GPCRs that lead to degranulation and the regulation of these pathways during the degranulation response are areas of active study. This review summarizes the current understanding of the mechanisms involved in the regulation of degranulation through GPCRs.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1574-6968.2004.tb09619.x
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