Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound   Target Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound

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Query: EC:6.3.4.6 (urease)
7,490 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Pyrolysis gas chromatography (PGC) has been shown to be useful for differentiating enzymes. The enzymes alpha-chymotrypsin, lactate dehydrogenase, catalase, and urease were easily "fingerprinted" on a 1.8 m 0.5% Carbowax 20 M column. Also, in some cases, isoenzymes of lactate dehydrogenase could be distinguished. Based on the pyrolyses of the free aromatic amino acids, four major enzyme pyrolysis peaks were tentatively identified as organic compounds derived from tyrosine and tryptophan. The use of a nitrogen-selective detector in conjunction with the FID and measurement of peak retention times by computer on three different types of columns permitted confirmations of peak identity.
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PMID:Pyrolysis gas chromatography of enzymes. 73 Aug 12

Cysteine 319 in the large subunit of Klebsiella aerogenes urease was identified as an essential catalytic residue based on chemical modification studies (Todd, M.J., and Hausinger, R.P. (1991) J. Biol. Chem. 266, 24327-24331). Through site-directed mutagenesis, this cysteine has been changed independently to alanine, serine, aspartate, and tyrosine. None of these mutations (C319A, C319S, C319D, and C319Y, respectively) affected the size or level of synthesis of the urease subunits as monitored by polyacrylamide gel electrophoresis. The wild type enzyme and each of the mutant proteins was purified and their properties were compared. The C319Y protein possessed no detectable activity, while activity was reduced in C319A, C319S, and C319D to 48, 4.5, and 0.03% of wild type levels under normal assay conditions. All of the active mutants had a small increase in Km when compared to the wild type value. The active mutants displayed a greatly reduced sensitivity to inactivation by iodoacetamide in comparison to the wild type enzyme, confirming our previous assignment of the essential cysteine to this residue based on active site peptide mapping. In contrast to the wild type enzyme, inactivation of the mutant proteins was not affected by the presence of the competitive inhibitor phosphate, suggesting that the remaining slow rate of iodoacetamide inactivation is due to modification away from the active site. The pH dependence of urease activity was substantially altered in the active mutants with C319S and C319D showing a pH optimum near 5.2, and C319A near 6.7, compared to the pH 7.75 optimum of wild type urease. These data are consistent with Cys-319 facilitating catalysis at neutral and basic pH values by participating as a general acid.
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PMID:Site-directed mutagenesis of the active site cysteine in Klebsiella aerogenes urease. 140 Mar 17

A modification of the procedure for O-1 phage Salmonella screening is presented. The novel method is based on the use of two media, i.e., a new medium (double sugar-tyrosine [DST]), which permits the combination of adonitol and sucrose fermentation and tyrosine clearing tests, and the previously described o-nitrophenyl-beta-D-galactopyranoside urease indole medium. In comparative trials, the new procedure and the conventional one were used to screen for Salmonella isolates from 553 lactose-negative strains of members of the family Enterobacteriaceae. The O-1 phage test, performed on DST medium, recognized the same number of phage-susceptible Salmonella strains as did the standardized method; however, it permitted the correct identification of a greater number of phage-resistant strains for discard (95.6 versus 85.3%). In particular, DST medium presented a higher efficacy than triple sugar iron agar (which is the corresponding medium in the reference procedure) in correctly identifying phage-negative cultures for discard (69.1 versus 28.5%).
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PMID:Double sugar-tyrosine medium improves O-1 phage Salmonella screening. 153 32

Forty one strains of dematiaceous fungi from the Mycology collection of the University of Costa Rica were studied. Thirty three were pathogenic (Fonsecaea pedrosoi, Cladosporium carrionii, Xylohypha bantiana, Exophiala jeanselmei, Rhinocladiella aquaspersa, Phialophora verrucosa) and the other eight were contaminants (Hormodendrum sp.). Morphological studies were done using the slide culture technique. The physiological criteria used were: urease production, gelatin and Loeffler media liquefaction; xanthine, tyrosine, starch and casein hydrolysis; nitrate utilization; carbohydrate uptake; sensitivity to cycloheximide and thermotolerance in glucose-Sabouraud medium. The physiological tests did not provide characteristic patterns for the different genera of pathogenic fungi, even though these differences were detected in non pathogenic fungi; the tests may be useful for the quick separation of both groups. Physiological test may have a limited value in the identification of fungi and the morphological analysis cannot be substituted by physiological studies.
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PMID:[Morphologic and physiologic characteristics of Costa Rica pathogenic fungi (Dermatiaceae)]. 184 48

Three cases of great toenail infection are described in which a slow-growing arthroconidial hyphomycete was isolated repeatedly and in pure culture. Direct microscopy revealed hyaline, round to barrel-shaped arthroconidia, hyaline hyphae of varying width, and broad thick-walled brownish hyphae. Three additional isolates were obtained from clinical specimens, for which the results of direct microscopy were unknown or negative. The fungus was resistant to cycloheximide, sensitive to common antifungal drugs by susceptibility tests in vitro and sensitive to benomyl. It was urease positive, hydrolysed casein and tyrosine but not xanthine or hypoxanthine, showed no specific nutritional requirements but grew better on carbohydrate-free media, assimilated 12 carbohydrates and potassium nitrate, and failed to perforate hair. The fungus is described as Onychocola canadensis Sigler gen. et sp. nov., and it is compared to Scytalidium lignicola, Scytalidium hyalinum and the Scytalidium synanamorph of Nattrassia mangiferae (Hendersonula toruloidea).
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PMID:Toenail infection caused by Onychocola canadensis gen. et sp. nov. 214 86

The effects of oral and intraperitoneal administration of biotin in urease-induced hyperammonemic rats, as well as the influence of biotin deficiency, have been studied. Biotin deficiency was produced by feeding standard diet MF (Oriental Yeast Co.) supplemented with dry egg-white (egg-white group). Egg-white + biotin group had free access to 0.0014% of biotin solution at all time. Following an intraperitoneal injection of urease, 25 U/kg (B.W.), plasma ammonia levels in egg-white + biotin group were lower than in egg-white group, especially there was significance (p less than 0.05) at 8 hours after the urease injection. Similarly, plasma ammonia levels in biotin-injected rats, in which 1 mg of biotin had been injected intraperitoneally prior to the experiment, were significantly low compared with saline-injected controls at 4 and 6 hours after urease administration. Results of plasma amino acid analysis, 9 hours after the urease injection indicated that Fischer's molar ratio (Leu + Ileu + Val/Tyr + Phe) was significantly higher in the biotin-injected rats than the saline-injected control. It suggests that biotin might decrease blood ammonia by facilitating the detoxification mechanism as follow: L-glutamate + NH3----L-glutamine.
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PMID:[The effects of biotin on the metabolism of ammonia and amino acids in urease-induced hyperammonemic rats]. 281 Aug 55

Scolecobasidium humicola, a soil fungus and etiologic agent of phaeohyphomycosis in fish, is herein reported to cause cutaneous lesions in a tortoise, Terrapine carolina var. carolina. S. humicola was isolated from lesions on the foot and dematiaceous hyphae were observed in KOH preparations of the biopsy and in stained preparations. This isolate and others were compared morphologically and physiologically with isolates of Dactylaria gallopava which it resembles. As a result of this investigation, we concluded that D. gallopava may be differentiated from S. humicola macroscopically, by the production in D. gallopava of an extensive diffusible purplish-red to reddish-brown pigment when cultured on Sabouraud dextrose agar; microscopically, by the presence and usually predominance of conidia, whose apical cell is markedly wider than the basal cell, and usually constricted at the septum; and physiologically, by the ability to grow on media containing cycloheximide and by the ability to grow well at 36-45 degrees C. In contrast, S. humicola does not usually produce a diffusible pigment on Sabouraud's dextrose agar or if present, is not extensive; it lacks the wider upper cell; is less constricted or non-constricted at the central septum; grows on media containing cycloheximide, although some inhibition may occur and lastly, does not grow at 36 degrees C or higher. Both species were urease positive, hydrolysed tyrosine but not casein, xanthine, or gelatin.
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PMID:A comparison between Dactylaria gallopava and Scolecobasidium humicola: first report of an infection in a tortoise caused by S. humicola. 404 87

1. Urease of specific activity 160-180 Sumner units/g. (Sumner, 1951) was purified from jack-bean meal. The preparation was pure on the basis of polyacryl-amide-gel electrophoresis and N-terminal studies. 2. By using both the 1-fluoro-2,4-dinitrobenzene method and the phenyl isothiocyanate method a single N-terminal methionine residue was found. 3. A single C-terminal sequence -Tyr-Leu-Phe was found by studies with carboxypeptidase A, carboxypeptidase B and hydrazinolysis. 4. N-Bromosuccinimide cleavage showed that five unique tryptophan sequences were present: Trp-Ala, Trp-Glu, Trp-Gly, Trp-Met and Trp-Arg. 5. Polyacrylamide-gel electrophoresis in sodium dodecyl sulphate showed that urease had a subunit molecular weight of 76000. 6. The yield of N- and C-terminal amino acids, the number of tryptic peptides and tryptophan sequences and the above polyacrylamide-gel electrophoretic measurement all suggest that urease contains a single structural subunit of molecular weight 75000.
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PMID:The subunit structure of jack-bean urease. 538 87

Monoconidial cultures derived from 12 clinical and environmental isolates of Phialophora parasitica were compared with respect to morphologic and physiologic characteristics and response to antifungal agents. No yeast cells were seen in 1- and 3-week-old slide culture preparations. Also, not all of the distinguishing characteristics of this species were displayed by all isolates on all media examined. Although the isolates grew on Sabouraud agar with chloramphenicol and cycloheximide, some inhibition was observed. All cultures were strongly urease-positive and hydrolyzed casein and starch; most decomposed tyrosine but not gelatin. All but one environmental isolate grew well at both 23 and 37 degrees C, but none grew at 40 degrees C. In the sensitivity testing the isolates did not vary much in their response to each drug, although some anomalies were observed. Amphotericin B and miconazole had minimum inhibitory concentrations in the low sensitivity range (2.0-8.0 and 2.5-10 micrograms m-1 respectively), for most isolates, and most isolates were resistant to both 5-fluorocytosine and ketoconazole. Limited observations were made on three other Phialophora species which might be confused with P. parasitica.
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PMID:Phialophora parasitica, an emerging pathogen. 609 71

A study, conducted with 20 isolates of Microsporum ferrugineum and 12 isolates of Trichophyton soudanense, revealed that some of the discrepancies in the literature regarding their characteristics and differentiation were due to methodology, strain variation and the use of an insufficient number of isolates. We found all isolates of T. soudanense to be urease negative and gelatinase positive (usually by the first week); to produce brown to black colonies on Lowenstein-Jensen medium; to rapidly decompose casein and more slowly tyrosine; to grow well or better at 37 degrees C as compared to room temperature; to produce reflexive branching on cornmeal Tween agar and abundant microconidia on casero medium and to exhibit no sexual reaction with either mating type of arthroderma simii. All but one isolate demonstrated restricted growth on lactose agar and only three isolates perforated hair. In contrast, we found 18 of 20 isolates of M. ferrugineum to be urease positive in urea broth (most isolates were negative on urea agar); all produced light-colored colonies on Lowenstein-Jensen medium; spreading colonies on lactose agar and failed to perforate hair in vitro or to produce reflexive branching. Most isolates manifested poorer to no growth at 37 degrees C compared to room temperature and all but one failed to decompose casein and tyrosine. A few strains produced macroconidia and/or microconidia on casero medium and some reacted sexually with A. simii (a) (-) mating type. Gelatin hydrolysis was variable.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Studies in the differentiation between Microsporum ferrugineum Ota and Trichophyton soudanense Joyeux. 637 42


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