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uricase of bacillus fastidiosus. properties and regulation of synthesis.uricase (urate:oxygen oxidoreductase, ec 1.7.3.3) of bacillus fastidiosus was purified to homogeneity in a two-step procedure and was crystallized. the native molecule had a molecular weight of 145 000--150 000 and was composed of subunits of two kinds (mr = 36 000 and 39 000) in a 1 : 1 ratio. the quaternary structure of the enzyme was reversibly altered, with concomitant loss of activity, at temperatures between 40 and 60 degrees c. no evidence was found for the involvement of metal ions or co ...1978728443
purification and characterization of the nadp-dependent glutamate dehydrogenase from bacillus fastidiosus.ammonia assimilation in bacillus fastidiosus proceeds via the nadp-dependent glutamate dehydrogenase. the enzyme, purified to homogeneity, is composed of identical subunits with a molecular weight of about 48,000 dalton. presumably the enzyme is a hexamer. the enzyme is specific for nadp(h). the ph optima for the amination and deamination reactions are 7.7 and 8.6, respectively. the temperature optimum is 60 degrees c. furthermore, temperature stability and apparent km values for substrates of b ...19892543290
role of uricase in the triggering of germination of bacillus fastidiosus spores.the likelihood that uric acid was the only compound capable of triggering germination of bacillus fastidiosus spores was reinforced by the finding that ureidoglycollic acid, urea, nh4cl, 2,8-dihydroxypurine and a combination of l-alanine and o-carbamoyl-d-serine were ineffective as germinants. uric acid-triggered germination of b. fastidiosus was prevented by a range of inhibitors that also inhibited uricase activity in dormant spore extracts. o2 uptake during germination started immediately aft ...19854038258
stimulation of growth of bacillus fastidiosus by amino acids.growth of bacillus fastidiosus on allantoin is stimulated by components of rich media as was evident from a decreased generation time and an increased maximal growth yield in the presence of such media. such a stimulation must involve various transport systems. energy-dependent transport systems for some amino acids were demonstrated besides those for urate, allantoin and allantoate.19846465879
purification and characterization of allantoate amidohydrolase from bacillus fastidiosus.allantoate amidohydrolase from bacillus fastidiosus was purified 170-fold to homogeneity as judged by isoelectric focusing and nondenaturing and sodium dodecyl sulfate polyacrylamide gel electrophoresis. the molecular mass was estimated to be 128 kda. the enzyme appeared to be a homodimer with a subunit molecular mass of 66 kda. the enzyme has an isoelectric point of 5.6. allantoate amidohydrolase is a mn(2+)-dependent enzyme exhibiting a ph optimum around 8.8. its km value for allantoate was es ...19957503567
studies on the physiology of bacillus fastidiosus.bacillus fastidiosus was grown in a minimal medium that contained uric acid or allantoin, aerated by vigorous stirring. a constant, optimum ph of 7.4 was maintained by controlled addition of sulfuric acid. washed cells converted both urate and allantoin into carbon dioxide and ammonia, simultaneously assimilating part of the available carbon and nitrogen. urate oxidase (formerly called uricase) was present in extracts from urate-grown but not allantoin-grown cells. the formation of urate oxidase ...19715095289
inducement of a heat-shock requirement for germination and production of increased heat resistance in bacillus fastidiosus spores by manganous ions.bacillus fastidiosus, which requires uric acid or allantoin, grows and sporulates on a simple medium containing 59.5 mm uric acid, 5.7 mm k(2)hpo(4), and 2% agar in distilled water. seventy to ninety percent sporulation was achieved in 96 h. spores obtained on this medium do not need a heat shock prior to germination. the necessary germination conditions for this organism are 30 c, phosphate or this(hydroxymethyl)aminomethane buffer at ph 7.0, and 5.95 mm uric acid. sporulation occurred earlier ...19734698206
enhancing performance of uricase using multiwalled carbon nanotube doped polyaniline.multiwalled carbon nanotubes (cnt) doped polyaniline (pani) nanocomposite has been electrochemically deposited onto indium tin oxide (ito)-coated glass substrate for fabrication of uric acid biosensor. to achieve this, uricase (from bacillus fastidiosus) has been covalently immobilized onto glutaraldehyde-modified cnt-pani/ito and characterized using cyclic voltammetry (cv), scanning electron microscopy (sem), fourier transform infrared (ftir) spectroscopy, electrochemical impedance spectroscopy ...201424928549
a practical system for high-throughput screening of mutants of bacillus fastidiosus uricase.for high-throughput screening (hts) of bacillus fastidiosus uricase mutants, a practical system was proposed. by error-prone pcr with final 1.5 mm mncl2, two focused libraries of mutants for a1-v158 and v150-d212 were generated separately. after induced expression of individual clones in 48-well microplates, escherichia coli cells (bl21) were lyzed by 1.0 m tris-hcl at ph 9.0 in 96-well microplates at 25 °c for 7.5 ~ 10.5 h; uricase reaction was continuously monitored with 0.15 mm uric acid in 9 ...201727614625
effects of modification of amino groups with poly(ethylene glycol) on a recombinant uricase from bacillus fastidiosus.after modification with monomethoxyl-poly(ethylene glycol)-5000, a recombinant intracellular uricase from bacillus fastidiosus atcc 29604 showed residual activity of about 65%, a thermo-inactivation half-life >85 h, a circulating half-life about 20 h in rats in vivo, consistent effects of common cations, and consistent optima for reaction temperature and ph. thus, this uricase can be formulated via modification with monomethoxyl-poly(ethylene glycol).201020530883
reversible inactivation of an intracellular uricase from bacillus fastidiosus via dissociation of homotetramer into homodimers in solutions of low ionic strength.an intracellular uricase from bacillus fastidiosus with high catalytic capacity and strong resistance to xanthine was inactivated in water but could be essentially re-activated in solutions of high ionic strength. by polyacrylamide gel electrophoresis (page), gradient page, sodium-dodecyl-sulfate-page, gel-filtration through sephadex g200, and activity staining with peroxidase and its chromatogenic substrate, this homotetrameric uricase in water was found to dissociate into inactive homodimers t ...200919734651
bacillus herbersteinensis sp. nov.two bacterial strains, designated d-1,5a(t) and d-1,5b, were isolated from a medieval wall painting in the chapel of castle herberstein, styria (austria). the gram-positive, heterotrophic, aerobic, spore-forming rods showed nearly identical whole-cell protein patterns, identical genomic fingerprints and identical physiological profiles, demonstrating their relationship at the species level. both strains contained meso-diaminopimelic acid in their peptidoglycan, possessed a quinone system compris ...200516166719
epr spin trapping of a radical intermediate in the urate oxidase reaction.urate oxidase, or uricase (ec 1.7.3.3), is a peroxisomal enzyme that catalyses the oxidation of uric acid to allantoin. the chemical mechanism of the urate oxidase reaction has not been clearly established, but the involvement of radical intermediates was hypothesised. in this study epr spectroscopy by spin trapping of radical intermediates has been used in order to demonstrate the eventual presence of radical transient urate species. the oxidation reaction of uric acid by several uricases (porc ...200415571216
therapeutic proteins: a comparison of chemical and biological properties of uricase conjugated to linear or branched poly(ethylene glycol) and poly(n-acryloylmorpholine).uricase from bacillus fastidiosus (uc) was covalently linked to linear peg (peg-1) (mw 5 kda), branched peg (peg-2) (mw 10 kda) and to poly(n-acryloylmorpholine) (pacm) (mw 6 kda). the conjugation of uc with linear peg and pacm was accompanied by complete loss of enzymatic activity but, if uric acid as site protecting agent was included in the reaction mixture, the conjugate protein retained enzymatic activity. on the other hand, the modification with peg-2 gave a conjugate that also maintained ...200010966157
the level of enzymes involved in the allantoin metabolism of bacillus fastidiosus grown under different conditions.bacillus fastidiosus was cultivated in batch and continuous culture on various carbon and nitrogen sources. the enzymes involved in allantoin degradation (allantoinase, urease, carboligase) of b. fastidiosus were hardly affected by either carbon or nitrogen source. in contrast, the enzymes involved in glycerol utilization (glycerol kinase, glycerol 3-phosphate dehydrogenase) were induced during growth on glycerol, but were not affected by the amount of allantoin present.19957765882
purification and some properties of hydroxypyruvate isomerase of bacillus fastidiosus.hydroxypyruvate isomerase of bacillus fastidiosus is a novel enzyme (braun, w. and kaltwasser, h. (1979) arch. microbiol. 121, 129-134) which catalyzes the reversible conversion of tartronate semialdehyde into hydroxypyruvate. the enzyme was purified to homogeneity. the native molecule had a molecular weight of 265 000-280 000 and was composed of six subunits with a molecular weight of 45 000. the enzyme showed optimal activity at ph 6.6-7.4 and 57 degrees c. hydroxypyruvate isomerase is stable ...19807448201
uric acid degradation by bacillus fastidiosus strains.seven bacillus strains including one of the original bacillus fastidiosus strains of den dooren de jong could grow on urate, allantoin, and, except one, on allantoate. no growth could be detected on adenine, guanine, hypoxanthine, xanthine, and on degradation products of allantoate. some strains grew very slowly in complex media. the metabolic pathway from urate to glyoxylate involved uricase, s(+)-allantoinase, allantoate amidohydrolase, s(-)-ureidoglycolase, and, in some strains, urease.19761245468
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