Forgot your password?
New user?
New Window Opens on the Secret Life of Microbes: Scientists Develop First Microbial Profiles of Ecosystems
CAS announces the release of SciFinder Scholar for Mac OS X
Press Releases
Press Releases
| Check out our New Publishers' Select for Free Articles |
Archives of Microbiology (v.176, #5)
No Title by Yoichi Ogawa; Masahiro Mii (pp. 315-322).
The genetic similarity between pTiBo542 and pTiCNI5, which are harbored, respectively by the supervirulent Agrobacterium tumefaciens strain A281 and by the highly tumorigenic wild-type strain CNI5 isolated from chrysanthemum was investigated by Southern hybridization. pTiCNI5 and pTiBo542 exhibited highly similar hybridization patterns in both BamHI- and EcoRI-digested plasmids by using four vir-region-specific probes, whereas similarity in these two plasmids was not observed by probing with five TL-DNA-specific probes. The characteristics related to tumor formation of cryptic plasmids carried by strain CNI5 were investigated by using single-plasmid-cured derivatives. pTiCNI5-cured derivatives predictably failed to utilize agropine and mannopine and failed to induce tumors on chrysanthemum and tobacco leaf explants, while pAtCNI5a-, pAtCNI5c- and pAtCNI5d-cured derivatives could utilize these opines similar to the parent strain CNI5. Interestingly, pAtCNI5c- and pAtCNI5d-cured derivatives showed low tumorigenicity in comparison with strain CNI5 or with the pAtCNI5a-cured derivative. These results suggest that the highly virulent phenotype of strain CNI5 may be due to one or more vir genes, which exhibit cartographic similarity to those of pTiBo542. The results also suggest that the gene(s) related to tumor formation of strain CNI5 may exist not only on pTiCNI5 but also on cryptic plasmids pAtCNI5c and pAtCNI5d.
Keywords: Agrobacterium tumefaciens Chromosomal background Cryptic plasmid Vir genes T-DNA Ti plasmid Tumorigenicity
No Title by Ulysses Lins; Marcos Farina (pp. 323-328).
Magnetotactic multicellular aggregates consist of several bacteria that produce iron sulfide magnetosomes through a complex and poorly understood process. We observed new amorphous mineral particles within the cytoplasm of magnetotactic multicellular aggregates. Elemental mapping and electron energy loss spectroscopy detected iron and oxygen, but not sulfur, in these particles. These amorphous particles were about the same size as mature magnetosomes, around 50–70 nm in diameter. No membranes were observed surrounding the amorphous minerals. Partially crystalline inclusions composed of a crystalline core and an amorphous region around them similar in texture to the amorphous particles were also present. The shape of these amorphous regions followed the shape of the crystalline cores they enveloped. These regions also contained oxygen and iron. The crystalline phase, as previously reported, contained sulfur and iron. The presence of independent amorphous particles has not been reported before in magnetotactic multicellular aggregates.
Keywords: Magnetotactic bacteria Magnetosomes Iron sulfides Magnetotactic multicellular aggregates Biomineralization
No Title by Antje Labes; Peter Schönheit (pp. 329-338).
The hyperthermophilic, sulfate-reducing archaeon Archaeoglobus fulgidus strain 7324, rather than the type strain VC16, was found to grow on starch and sulfate as energy and carbon source. Fermentation products and enzyme activities were determined in starch-grown cells and compared to those of cells grown on lactate and sulfate. During exponential growth on starch, 1 mol of glucose-equivalent was incompletely oxidized with sulfate to approximately 2 mol acetate, 2 mol CO2 and 1 mol H2S. Starch-grown cells did not contain measurable amounts of the deazaflavin factor F420 (<0.03 nmol/mg protein) and thus did not show the F420-specific green-blue fluorescence. In contrast, lactate (1 mol) was completely oxidized with sulfate to 3 mol CO2 by strain 7324, and lactate-grown cells contained high amounts of F420 (0.6 nmol/mg protein). In extracts of starch-grown cells, the following enzymes of a modified Embden-Meyerhof pathway were detected: ADP-dependent hexokinase (ADP-HK), phosphoglucose isomerase, ADP-dependent 6-phosphofructokinase (ADP-PFK), fructose-1,6-phosphate aldolase, glyceraldehyde-3-phosphate:ferredoxin oxidoreductase (GAP:FdOR), phosphoglycerate mutase, enolase, and pyruvate kinase (PK). Specific activities of ADP-HK, ADP-PFK, GAP:FdOR, and PK were significantly higher in starch-grown cells than in lactate-grown cells, indicating induction of these enzymes during starch catabolism. Pyruvate conversion to acetate involved pyruvate:ferredoxin oxidoreductase and ADP-forming acetyl-CoA synthetase. The findings indicate that the archaeal sulfate reducer A. fulgidus strain 7324 converts starch to acetate via a modified Embden-Meyerhof pathway and acetyl-CoA synthetase (ADP-forming). This is the first report of growth of a sulfate reducer on starch, i.e. on a polymeric sugar.
Keywords: Hyperthermophiles Archaea Sulfate reducer Archaeoglobus fulgidus Sugar utilization Starch Modified Embden-Meyerhof pathway 5-Deazaflavin coenzyme F420 ADP-dependent hexokinase ADP-dependent 6-phosphofructokinase Glyceraldehyde-3-phosphate:ferredoxin oxidoreductase Acetyl-CoA synthetase (ADP-forming)
No Title by Ralf Geiben-Lynn; Klaus Sauber; Frieder Lutz (pp. 339-346).
φCTX is a double-stranded DNA phage of the Myoviridae family that converts Pseudomonas aeruginosa into a cytotoxin producer. A 42-kDa φCTX-inhibiting protein was purified from the outer membrane fraction of P. aeruginosa strain GuA18 by octyl-β-glucoside extraction, DEAE-chromatography, and mono-Q HPLC. This protein had an isoelectric point of 5.4 and bound specifically [125I]-labeled φCTX. The N-terminal amino acid sequence of six out of seven Lys-C fragments was highly similar (87%) to that of the entire of type-a flagellin of P. aeruginosa strain PAK. At a concentration of 14 nM, purified flagellin protein caused a 50% decrease in the phage titer after a 20-min incubation at 37 °C (PhI50). The presence of ethanol was necessary to reconstitute the inhibitory activity. In contrast, no ethanol treatment was necessary for the inhibitory activity of the sheared flagellin filaments from P. aeruginosa strain GuA18, which consists of the 42-kDa flagellin subunits and the synthesized 17-mer phage-binding-peptide NGSNSDSERTALNGEAK, representing flagellin residues 100–116 of P. aeruginosa strain PAK. The PhI50 was 10 nM and 200 nM, respectively. Antisera against the flagellin filament protein as well as against the 17-mer peptide neutralized phage infection. These results indicated that the amino acid region 100–116 of the flagellin subunit of strain GuA18 is involved in φCTX binding. This region might play a role in phage attachment.
Keywords: Pseudomonas aeruginosa Phage φCTX Attachment site Flagellin
No Title by Claudia Schabereiter-Gurtner; Guadalupe Piñar; Dietmar Vybiral; Werner Lubitz; Sabine Rölleke (pp. 347-354).
A molecular approach was chosen to analyse the correlation between bacterial colonisation and rosy discolouration of masonry and lime wall paintings of two historically important buildings in Austria and Germany. The applied molecular method included PCR amplification of genes encoding the small subunit rRNA of bacteria (16S rDNA), genetic fingerprinting by denaturing gradient gel electrophoresis (DGGE), construction of 16S rDNA clone libraries, and comparative phylogenetic sequence analyses. The bacterial community of one red-pigmented biofilm sampled in Herberstein (Austria) contained bacteria phylogenetically related to the genera Saccharopolyspora, Nocardioides, Pseudonocardia, Rubrobacter, and to a Kineococcus-like bacterium. The bacterial community of the second red-pigmented biofilm sampled in Herberstein contained bacteria related to Arthrobacter, Comamonas, and to Rubrobacter. Rubrobacter-related 16S rDNA sequences were the most abundant. In the red-pigmented biofilm sampled in Burggen (Germany), only Rubrobacter-related bacteria were identified. No Rubrobacter-related bacteria were detected in non-rosy biofilms. The majority of sequences (70%) obtained from the bacterial communities of the three investigated rosy biofilms were related to sequences of the genus Rubrobacter (red-pigmented bacteria), demonstrating a correlation between Rubrobacter-related bacteria and the phenomenon of rosy discolouration of masonry and lime wall paintings.
Keywords: Denaturing gradient gel electrophoresis 16S rDNA Rosy biofilms Wall paintings and masonry Rubrobacter-related organism
No Title by Michael F. Dunn; Gisela Araíza; Turlough M. Finan (pp. 355-363).
The gene encoding pyruvate carboxylase (pyc) was isolated from a Sinorhizobium meliloti Rm1021 cosmid bank by complementation of a Rhizobium tropici pyc mutant. PYC-negative mutants of S. meliloti Rm1021 were isolated by transposon mutagenesis and were unable to grow with glucose or pyruvate as sole carbon sources, but were symbiotically competent in combination with alfalfa plants. PYC activity assays, pyc::lacZ gene fusion studies and an in vivo biotinylation assay showed that PYC activity in S. meliloti was dependent mainly on biotin availability and not on changes in gene transcription. The subunit and holo-enzyme molecular masses of the S. meliloti PYC indicated that the enzyme was an α4 homotetramer. The S. meliloti PYC had a high apparent K a (0.23 mM) for the allosteric activator acetyl-CoA and was product-inhibited by sub-millimolar concentrations of oxaloacetate. In contrast to other bacterial α4-PYCs which have been characterized, the S. meliloti enzyme was not strongly inhibited by L-aspartate.
Keywords: Pyruvate carboxylase Anaplerotic reactions Carbon metabolism Rhizobia Sinorhizobium meliloti Biotin
No Title by Akihiro Hara; Mami Arie; Tamotsu Kanai; Toru Matsui; Hitoshi Matsuda; Keizo Furuhashi; Mitsuyoshi Ueda; Atsuo Tanaka (pp. 364-369).
We established a novel and convenient method to construct a ura3 strain (ura3/ura3) of the asporogenous and diploid yeast, Candida tropicalis, that produces dicarboxylic acid. One copy of the URA3 gene was disrupted using a mutated hygromycin B resistance gene (HYG#). The obtained hygromycin-resistant strain was further transformed with a URA3 disruption cassette and selected on a plate containing 5-fluoroorotic acid. The obtained strains were analyzed and the disruption of the gene was confirmed by PCR and Southern blot analysis. The results showed that the strains were obtained in which allelic URA3 genes were simultaneously disrupted. Furthermore, we established a cotransformation method for this gene disruption, using HYG# in C. tropicalis. In order to disrupt the allelic POX4 genes (encoding acyl-CoA oxidase) of dicarboxylic acid-producing strains, the ARS plasmid (which contained HYG#) and a POX4 disruption cassette (which carried the LAC4 gene encoding β-galactosidase of Kluyveromyces lactis) were simultaneously introduced by transformation. As a result, the allelic POX4 gene was successfully disrupted.
Keywords: Hygromycin B Candida tropicalis Acyl-CoA oxidase POX4 Dicarboxylic acid n-Alkane Gene disruption
No Title by Sally M. Hoffer; Nathalie Uden; Jan Tommassen (pp. 370-376).
The uptake of hexose 6-phosphates in Escherichia coli is mediated by the transporter UhpT, the synthesis of which is induced by the presence of glucose 6-phosphate (glucose 6-P) in the medium. Since this protein functions as an anion exchanger, it is generally assumed to be geared for the use of sugar phosphates as a carbon source. However, the question was unresolved whether this transporter can also provide the cells with glucose 6-P as a phosphate source. It is demonstrated in this work that UhpT-mediated glucose 6-P uptake does not allow the cells to grow on glucose 6-P as phosphate source. Hence, the expression of UhpT under phosphate limitation would not be particularly advantageous and some form of interaction between the uhp system and the Pi-limitation-inducible pho regulon, the products of which are involved in phosphate acquisition, may be anticipated. Indeed, the use of an uhpT-lacZ fusion revealed that much higher concentrations of the inducer glucose 6-P were required to elevate uhpT transcription when the pho regulon was expressed. This interference was the result of degradation of glucose 6-P by one of the products of the pho regulon, the periplasmic enzyme alkaline phosphatase. The specific form of interaction between the Pho and Uhp systems is designated inducer degradation.
Keywords: uhp genes pho regulon Inducer degradation Escherichia coli
No Title by Nick Geukens; Victor Parro; Luis A. Rivas; Rafael P. Mellado; Jozef Anné (pp. 377-380).
Type I signal peptidases are responsible for the proteolytic cleavage of the signal peptide of secreted proteins. In the gram-positive bacterium Streptomyces lividans, four adjacent genes (sipW, sipX, sipY and sipZ) were isolated encoding putative type I signal peptidases. In this work, the different sip genes were cloned and expressed. Subsequently, the Sip proteins were purified to raise antibodies. Although the four Sip proteins share a low degree of sequence similarity and differ significantly in size and pI, anti-Sip antibodies cross-reacted intensively. Functional signal peptidase processing activity for each of these Sip proteins was shown both in vitro and in vivo. The different Sip proteins did not exhibit the same cleavage efficiency on the Bacillus subtilis pre-chitosanase.
Keywords: Signal peptidase Streptomyces Protein secretion
No Title by F. Rafii; G. Hehman; P. Lunsford (pp. 381-385).
Mycobacterium sp. Pyr-1 produces an enzyme with nitroreductase activity that reduces 1-nitropyrene and 4-nitrobenzoic acid to the corresponding aromatic amines. This enzyme was constitutive and required NADH; and its activity was enhanced by FAD. It was inhibited by antimycin A, dicumarol, and o-iodosobenzoic acid; and it was inactivated by ammonium sulfate precipitation. After purification to homogeneity, the protein produced a single band on native and SDS-polyacrylamide gels and had a single amino-terminal sequence. The N-terminal amino acid sequence was identical to the corresponding sequences of the lipoamide dehydrogenases of M. leprae, M. tuberculosis and Corynebacterium glutamicum. The amino-terminal sequence was also similar to lipoamide dehydrogenases from M. smegmatis and several other bacteria. The amino acid sequence of an internal peptide (12 of 13 amino acids) was nearly identical to the corresponding sequences of lipoamide dehydrogenases from M. leprae and M. tuberculosis and was similar to those of C. glutamicum, Streptomyces coelicolor and S. seoulensis. The data show that a unique lipoamide dehydrogenase in Mycobacterium sp. Pyr-1, which differs from classic (Type I) bacterial nitroreductases, reduces aromatic nitro compounds to aromatic amines.
Keywords: Mycobacterium Nitroreductase ADEPT Lipoamide dehydrogenase Nitro compounds
No Title by Milind G. Watve; Rashmi Tickoo; Maithili M. Jog; Bhalachandra D. Bhole (pp. 386-390).
Streptomyces is the largest antibiotic-producing genus in the microbial world discovered so far. The number of antimicrobial compounds reported from the species of this genus per year increased almost exponentially for about two decades, followed by a steady rise to reach a peak in the 1970s, and with a substantial decline in the late 1980s and 1990s. The cumulative number shows a sigmoid curve that is much flatter than what a logistic equation would predict. We attempted to fit a mathematical model to this curve in order to estimate the number of undiscovered antimicrobials from this genus as well as to predict the trends in the near future. A model assuming that the screening efforts are encouraged by a previous year's success and that the probability of finding a new antibiotic is a function of the fraction of antibiotics undiscovered so far offered a good fit after optimizing parameters. The model estimated the total number of antimicrobial compounds that this genus is capable of producing to be of the order of a 100,000 – a tiny fraction of which has been unearthed so far. The decline in the slope appeared to be due to a decline in screening efforts rather than an exhaustion of compounds. Left to itself, the slope will become zero in the next one or two decades, but if the screening efforts are maintained constant, the rate of discovery of new compounds will not decline for several decades to come.
Keywords: Actinomycetes Antibiotic diversity Drug discovery Drug screening Logistic model Mathematical model Secondary metabolites Streptomyces
No Title
by Bram A. Pas; Hermie J. Harmsen; Gerwin C. Raangs; Willem M. Vos; Gosse Schraa; Alfons J. Stams (pp. 391-392).