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Journal of Bacteriology Mar 19976-Phosphoryl-beta-D-glucopyranosyl:6-phosphoglucohydrolase (P-beta-glucosidase, EC 3.2.1.86) has been purified from Fusobacterium mortiferum. Assays for enzyme activity...
6-Phosphoryl-beta-D-glucopyranosyl:6-phosphoglucohydrolase (P-beta-glucosidase, EC 3.2.1.86) has been purified from Fusobacterium mortiferum. Assays for enzyme activity and results from Western immunoblots showed that P-beta-glucosidase (Mr, 53,000; pI, 4.5) was induced by growth of F. mortiferum on beta-glucosides. The novel chromogenic and fluorogenic substrates, p-nitrophenyl-beta-D-glucopyranoside-6-phosphate (pNPbetaGlc6P) and 4-methylumbelliferyl-beta-D-glucopyranoside-6-phosphate (4MUbetaGlc6P), respectively, were used for the assay of P-beta-glucosidase activity. The enzyme hydrolyzed several P-beta-glucosides, including the isomeric disaccharide phosphates cellobiose-6-phosphate, gentiobiose-6-phosphate, sophorose-6-phosphate, and laminaribiose-6-phosphate, to yield glucose-6-phosphate and appropriate aglycons. The kinetic parameters for each substrate are reported. P-beta-glucosidase from F. mortiferum was inactivated by 6-phosphoglucono-delta-lactone (P-glucono-delta-lactone) derived via oxidation of glucose 6-phosphate. The pbgA gene that encodes P-beta-glucosidase from F. mortiferum has been cloned and sequenced. The first 42 residues deduced from the nucleotide sequence matched those determined for the N terminus by automated Edman degradation of the purified enzyme. From the predicted sequence of 466 amino acids, two catalytically important glutamyl residues have been identified. Comparative alignment of the amino acid sequences of P-beta-glucosidase from Escherichia coli and F. mortiferum indicates potential binding sites for the inhibitory P-glucono-delta-lactone to the enzyme from F. mortiferum.
Topics: Amino Acid Sequence; Binding Sites; Cloning, Molecular; Fusobacterium; Gluconates; Glucosephosphate Dehydrogenase; Glucosidases; Glucosides; Kinetics; Lactones; Molecular Sequence Data; Molecular Weight; Sequence Alignment; Substrate Specificity
PubMed: 9045824
DOI: 10.1128/jb.179.5.1636-1645.1997 -
Journal of the National Medical... Apr 1986A case of anaerobic sepsis associated with Fusobacterium mortiferum is reported. Blood cultures from a 60-year-old man with type II diabetes mellitus, hypertension,...
A case of anaerobic sepsis associated with Fusobacterium mortiferum is reported. Blood cultures from a 60-year-old man with type II diabetes mellitus, hypertension, severe atherosclerotic cardiovascular disease, and renal insufficiency revealed on a gramstained smear highly pleomorphic gram-negative bacilli with bizarre forms and round bodies. Growth of the organism on nonselective anaerobic media and analysis of its pattern produced results characteristic of Fusobacterium mortiferum.
Topics: Fusobacterium Infections; Humans; Male; Middle Aged; Periodontitis; Sepsis
PubMed: 3712472
DOI: No ID Found -
Infection and Immunity Dec 1973The effects of Bacteroides sp., Fusobacterium mortiferum, Bacteroides fragilis, and Sphaerophorus necrophorus on various parameters of blood coagulation in vivo and in... (Comparative Study)
Comparative Study
The effects of Bacteroides sp., Fusobacterium mortiferum, Bacteroides fragilis, and Sphaerophorus necrophorus on various parameters of blood coagulation in vivo and in vitro were determined and compared to the coagulation effects of Escherichia coli and Salmonella minnesota, wild type and R595. Intravenous injection of washed cells, culture filtrate, lipopolysaccharide, or lipid A of the anaerobic gram-negative microorganisms into mice resulted in acceleration of coagulation. Lipopolysaccharide and lipid A of the anaerobic microorganisms had no apparent effect on circulating platelets in mice or rabbits and did not cause aggregation of human platelets in vitro. Washed cells, lipopolysaccharide, and lipid A of Bacteroides sp. and F. mortiferum also significantly accelerated the clotting time of recalcified platelet poor normal human plasma and C6-deficient rabbit plasma. Lipid A, but not lipopolysaccharide, of E. coli and washed cells of S. minnesota R595 accelerated coagulation by a similar mechanism. These results indicated that Bacteroides sp. and F. mortiferum can accelerate blood coagulation in vivo and in vitro by a mechanism which does not involve platelets or terminal components of complement.
Topics: Animals; Bacteroides; Blood Coagulation; Blood Coagulation Tests; Blood Platelets; Cell Wall; Escherichia coli; Female; Fusobacterium; Humans; Injections, Intravenous; Leukocyte Count; Lipids; Lipopolysaccharides; Mice; Platelet Adhesiveness; Polysaccharides, Bacterial; Rabbits; Salmonella; Salmonella enteritidis; Species Specificity; Thrombophlebitis; Time Factors
PubMed: 4594118
DOI: 10.1128/iai.8.6.911-918.1973 -
Journal of Clinical Microbiology May 2003Due to the inadequate automation in the amplification and sequencing procedures, the use of 16S rRNA gene sequence-based methods in clinical microbiology laboratories is...
Usefulness of the MicroSeq 500 16S ribosomal DNA-based bacterial identification system for identification of clinically significant bacterial isolates with ambiguous biochemical profiles.
Due to the inadequate automation in the amplification and sequencing procedures, the use of 16S rRNA gene sequence-based methods in clinical microbiology laboratories is largely limited to identification of strains that are difficult to identify by phenotypic methods. In this study, using conventional full-sequence 16S rRNA gene sequencing as the "gold standard," we evaluated the usefulness of the MicroSeq 500 16S ribosomal DNA (rDNA)-based bacterial identification system, which involves amplification and sequencing of the first 527-bp fragment of the 16S rRNA genes of bacterial strains and analysis of the sequences using the database of the system, for identification of clinically significant bacterial isolates with ambiguous biochemical profiles. Among 37 clinically significant bacterial strains that showed ambiguous biochemical profiles, representing 37 nonduplicating aerobic gram-positive and gram-negative, anaerobic, and Mycobacterium species, the MicroSeq 500 16S rDNA-based bacterial identification system was successful in identifying 30 (81.1%) of them. Five (13.5%) isolates were misidentified at the genus level (Granulicatella adiacens was misidentified as Abiotrophia defectiva, Helcococcus kunzii was misidentified as Clostridium hastiforme, Olsenella uli was misidentified as Atopobium rimae, Leptotrichia buccalis was misidentified as Fusobacterium mortiferum, and Bergeyella zoohelcum was misidentified as Rimerella anatipestifer), and two (5.4%) were misidentified at the species level (Actinomyces odontolyticus was misidentified as Actinomyces meyeri and Arcobacter cryaerophilus was misidentified as Arcobacter butzleri). When the same 527-bp DNA sequences of these seven isolates were compared to the known 16S rRNA gene sequences in the GenBank, five yielded the correct identity, with good discrimination between the best and second best match sequences, meaning that the reason for misidentification in these five isolates was due to a lack of the 16S rRNA gene sequences of these bacteria in the database of the MicroSeq 500 16S rDNA-based bacterial identification system. In conclusion, the MicroSeq 500 16S rDNA-based bacterial identification system is useful for identification of most clinically important bacterial strains with ambiguous biochemical profiles, but the database of the MicroSeq 500 16S rDNA-based bacterial identification system has to be expanded in order to encompass the rarely encountered bacterial species and achieve better accuracy in bacterial identification.
Topics: Bacteria; Bacterial Typing Techniques; Base Sequence; DNA Primers; DNA, Bacterial; DNA, Ribosomal; Humans; Polymerase Chain Reaction; RNA, Bacterial; RNA, Ribosomal, 16S
PubMed: 12734240
DOI: 10.1128/JCM.41.5.1996-2001.2003 -
Antimicrobial Agents and Chemotherapy Jul 2003By using an agar dilution method, the in vitro activities of ramoplanin, teicoplanin, vancomycin, linezolid, and five other agents were determined against 300...
By using an agar dilution method, the in vitro activities of ramoplanin, teicoplanin, vancomycin, linezolid, and five other agents were determined against 300 gram-positive and 54 gram-negative strains of intestinal anaerobes. Ramoplanin was active at
Clostridium difficile for which MICs of ramoplanin were 0.25 to 0.5 microg/ml; for 3 of these, linezolid MICs were 8 to 16 micro g/ml. Nineteen Clostridium innocuum strains for which the vancomycin MIC at which 90% of strains were inhibited was 16 microg/ml were susceptible to ramoplanin at 0.06 to 0.25 microg/ml and to teicoplanin at 0.125 to 1.0 microg/ml. All strains of Eubacterium, Actinomyces, Propionibacterium, and Peptostreptococcus spp. were inhibited by or=256 microg/ml. Ramoplanin displays excellent activity against C. difficile and other gram-positive enteric anaerobes, including vancomycin-resistant strains; however, it has poor activity against most gram-negative anaerobes and thus potentially has a lesser effect on the ecological balance of normal fecal flora. Topics: Acetamides; Actinomyces; Anti-Bacterial Agents; Bacitracin; Bacteria, Anaerobic; Clostridioides difficile; Depsipeptides; Eubacterium; Feces; In Vitro Techniques; Intestines; Linezolid; Microbial Sensitivity Tests; Oxazolidinones; Peptides, Cyclic; Peptostreptococcus; Propionibacterium; Teicoplanin; Vancomycin
PubMed: 12821492
DOI: 10.1128/AAC.47.7.2334-2338.2003 -
BMJ Case Reports Oct 2021Solitary fibrous tumours (SFTs) are rare mesenchymal tumours that are mostly seen in the pleura. Lately, they have also been described in other locations. Recent...
Solitary fibrous tumours (SFTs) are rare mesenchymal tumours that are mostly seen in the pleura. Lately, they have also been described in other locations. Recent discovery of the NAB2-STAT6 fusion gene which is specific for SFTs has led to an accurate diagnosis of SFTs. The occurrence of SFTs in the mesentery is very rarely reported in the literature. We report a case of a 63-year-old female who presented with abdominal pain, rectal bleeding and bacteraemia, who was ultimately found to have a mesenteric SFT.
Topics: Biomarkers, Tumor; Female; Fusobacterium; Humans; Immunohistochemistry; Mesentery; Middle Aged; Repressor Proteins; STAT6 Transcription Factor; Sepsis; Solitary Fibrous Tumors
PubMed: 34645630
DOI: 10.1136/bcr-2021-244603 -
Journal of Clinical Microbiology May 1979The survival of six species of anaerobic bacteria was studied in simple or commercially available diluents. Bacteroides fragilis and Fusobacterium nucleatum showed... (Comparative Study)
Comparative Study
The survival of six species of anaerobic bacteria was studied in simple or commercially available diluents. Bacteroides fragilis and Fusobacterium nucleatum showed excellent survival in all diluents including distilled water. Fusobacterium mortiferum survived well in all diluents except water and water supplemented with 0.1% gelatain. Clostridium perfringens survived best in phosphate-buffered saline with gelatin. Peptococcus asaccharolyticus required gelatin added to the basic diluent, and Streptococcus intermedius showed excellent survival only in minimal essential medium with gelatin. These diluents could provide effective and economical alternatives to more complex and costly diluents often used in work with anaerobic bacteria.
Topics: Anaerobiosis; Bacteria; Gelatin; Sodium Chloride; Solutions; Species Specificity; Water
PubMed: 479359
DOI: 10.1128/jcm.9.5.627-628.1979 -
Infection and Immunity Dec 1991Strains of eight Fusobacterium species differed in the ability to use sugars as energy sources for growth. For Fusobacterium russii ATCC 25533, F. gonidiaformans ATCC...
Strains of eight Fusobacterium species differed in the ability to use sugars as energy sources for growth. For Fusobacterium russii ATCC 25533, F. gonidiaformans ATCC 25563, and F. nucleatum ATCC 10953 (except for fructose), growth was marginal to poor on all of the sugars tested. Other species displayed reasonable growth on glucose, fructose, mannose, and galactose, and two strains of F. mortiferum (ATCC 25557 and ATCC 9817) grew well on six of the sugars tested, including sucrose and maltose. Glucose transport by resting cells of most of the species was dependent upon (or markedly stimulated by) the presence of a fermentable amino acid. By contrast, F. mortiferum cells rapidly accumulated glucose and other sugars in the absence of amino acids. Although these cells were constitutive for glucose uptake, accumulation of other sugars was specifically induced by growth of F. mortiferum on the appropriate sugar. Spectrophotometric analyses and in situ staining of anionic polyacrylamide gels showed that glucose and fructose (mannose) are phosphorylated by separate ATP-dependent kinases. Fructokinase was stable in air at 4 degrees C, but under these conditions, greater than 70% of the glucokinase activity was lost. After overnight dialysis of the extract, no glucokinase activity was detectable; however, 65% of the initial enzyme activity was retained by inclusion of 1 mM dithiothreitol in the dialysis buffer. Thin-section electron microscopy showed that cells of F. mortiferum produced various amounts of intracellular glycogen during growth on the following sugars (in decreasing order of formation): galactose greater than sucrose greater than glucose greater than mannose greater than fructose. Mechanisms for sugar transport regulation, phosphorylation, and polymer synthesis by F. mortiferum cells are proposed.
Topics: Biological Transport; Carbohydrate Metabolism; Fusobacterium; Glucose; Glycogen; Phosphorylation; Polysaccharides
PubMed: 1937813
DOI: 10.1128/iai.59.12.4547-4554.1991 -
Antimicrobial Agents and Chemotherapy Sep 1997HMR 3647 (RU 66647) and HMR 3004 (RU 64004), two ketolides, had MICs at which 50% of the strains are inhibited (MIC50s) of 0.06 to 0.125 microg/ml and MIC90s of 16.0... (Comparative Study)
Comparative Study
HMR 3647 (RU 66647) and HMR 3004 (RU 64004), two ketolides, had MICs at which 50% of the strains are inhibited (MIC50s) of 0.06 to 0.125 microg/ml and MIC90s of 16.0 microg/ml against 352 anaerobes. MIC50s and MIC90s of erythromycin, azithromycin, clarithromycin, and roxithromycin were 0.5 to 2.0 microg/ml and 32.0 to >64.0 microg/ml, respectively. HMR 3647 and HMR 3004 were more active against non-Bacteroides fragilis-group anaerobes (other than Fusobacterium mortiferum, Fusobacterium varium, and Clostridium difficile).
Topics: Anti-Bacterial Agents; Azithromycin; Bacteria, Anaerobic; Carbon Dioxide; Clarithromycin; Erythromycin; Hydrogen-Ion Concentration; Ketolides; Macrolides; Microbial Sensitivity Tests; Roxithromycin
PubMed: 9303406
DOI: 10.1128/AAC.41.9.2019 -
Journal of Bacteriology Jun 1994Phosphoenolypyruvate-dependent maltose:phosphotransferase activity was induced in cells of Fusobacterium mortiferum ATCC 25557 during growth on maltose. The disaccharide...
Phosphoenolypyruvate-dependent maltose:phosphotransferase activity was induced in cells of Fusobacterium mortiferum ATCC 25557 during growth on maltose. The disaccharide was rapidly metabolized by washed cells maintained under anaerobic conditions, but fermentation ceased immediately upon exposure of the cell suspension to air. Coincidentally, high levels of a phosphorylated derivative accumulated within the cells. Chemical and enzymatic analyses, in conjunction with data from 1H, 13C, and 31P nuclear magnetic resonance spectroscopy, established the structure of the purified compound as 6-O-phosphoryl-alpha-D-glucopyranosyl-(1-4)-D-glucose (maltose 6-phosphate). A method for the preparation of substrate amounts of this commercially unavailable disaccharide phosphate is described. Permeabilized cells of F. mortiferum catalyzed the phosphoenolpyruvate-dependent phosphorylation of maltose under aerobic conditions. However, the hydrolysis of maltose 6-phosphate (to glucose 6-phosphate and glucose) by permeabilized cells or cell-free preparations required either an anaerobic environment or addition of dithiothreitol to aerobic reaction mixtures. The first step in dissimilation of the phosphorylated disaccharide appears to be catalyzed by an oxygen-sensitive maltose 6-phosphate hydrolase. Cells of F. mortiferum, grown previously on maltose, fermented a variety of alpha-linked glucosides, including maltose, turanose, palatinose, maltitol, alpha-methylglucoside, trehalose, and isomaltose. Conversely, cells grown on the separate alpha-glucosides also metabolized maltose. For this anaerobic pathogen, we suggest that the maltose:phosphotransferase and maltose 6-phosphate hydrolase catalyze the phosphorylative translocation and cleavage not only of maltose but also of structurally analogous alpha-linked glucosides.
Topics: Aerobiosis; Anaerobiosis; Carbohydrate Sequence; Cell Membrane Permeability; Enzyme Induction; Fusobacterium; Glucosides; Hydrolysis; Maltose; Molecular Sequence Data; Phosphoenolpyruvate Sugar Phosphotransferase System; Phosphorylation; Stereoisomerism; Sugar Phosphates
PubMed: 8195080
DOI: 10.1128/jb.176.11.3250-3256.1994