• Asian-Australasian Journal of Animal Sciences
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• v.12 no.7
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• pp.1063-1069
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• 1999

Wheat straw samples (3-4 cm) were sprayed with solutions of urea (U) alone or with a dry addition of garden soil (GS), midden soil (MS), soya bean meal (SM) or jack bean meal (JM) as crude urease sources and with a pure urease (UR) enzyme. Each of the urease sources was included at two levels: 30 and 60 g/kg except pure urease, which was added at a level of 2.5 & 5.0 g/kg treated straw dry matter. Untreated straw without urease source was used as a control. After treatment, samples were sealed in polythene bags and stored for 2, 7, 14, 21 and 35 days at $19{^{\circ}C}$. The urease sources, their levels and treatment time produced significant effects on ammonia production (p<0.01). The addition of urease offered more flexibility in hydrolyzing urea in the shortest possible time. Incorporation of soya bean and jack bean meal was effective in reducing the modified acid detergent fiber (MADF) content of straw and the same time increasing organic matter (OM) digestibility. Overall effect, addition of soya bean to urea at a ratio of 1:1 appeared to be the most satisfactory urease source for the treatment of urea and wheat straw.

• Journal of Life Science
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• v.10 no.6
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• pp.558-563
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• 2000

Effect of inhibitors on Vibrio parahaemolyticus cell growth and its urease activity was studied. The growth of the bacterium and the enzyme activity were inhibited by the addition of 0.02% p-hydroxymercuric benzoate, $HgCl_2$and $AgNO_3$. However, same concentration of boric acid, thallium acetate and $Pb(NO_3)_2$ did not affect the cell growth but inhibited urease activity by 25%, 29%, and 38%, respectively. Acetohydroxamic acid was the most potent inhibitor on cell growth by inhibiting 40% but did not affect urease activity. To investigate the effect of inhibitors on urease activity, urease was purified and confirmed on SDS-PAGE. The purified urease was inhibited 100% by the addition of 1 mM acetohydroxamic acid and $AgNO_3$but no inhibition was occurred by the addition of the same concentration of thallium acetate. and the addition of 0.01 mM of $HgCl_2$ and acetohydroxamic acid inhibited the purified urease activity by 39% and 24%, respectively. On 0.1 millimolar basic, acetohydroxamic acid and $HgCl_2$inhibited 4 times more active in urease inhibition than p-hydroxymercuric benzoate whereas no inhibition was occurred either thallium acetate or $Pb(NO_3)_2$.

• BMB Reports
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• v.31 no.3
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• pp.240-244
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• 1998

Helicobacter pylori is the etiologic agent of human gastritis and peptic ulceration and produces urease as the major protein component on its surface. H. pylori urease is known to serve as a major virulence factor and in a potent immunogen. In order to express the recombinant urease at a higher level, a DNA fragment containing the minimal H. pylori urease gene cluster was subcloned into a high copy-number vector. The recombinant H. pylori urease expressed in an E. coli strain that was grown in a rich medium supplemented with added nickel was purified to near homogeneity by using DEAE-Sepharose, Superdex HR200, and Mono-Q (FPLC) columns and the purified enzyme possessed the specific activity of 1255 U/mg. Polyclonal antibodies raised against the purified recombinant H. pylori urease were shown to be very specific when subjected to Western blot analysis, in which crude extracts from the H. pylori ATCC strain and the recombinant E. coli strains expressing various bacterial ureases were exnmined for cross-reactivity.

• Biotechnology and Bioprocess Engineering:BBE
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• v.11 no.2
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• pp.140-145
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• 2006

The behaviour of alginate immobilized and soluble watermelon (Citrullus vulgaris) urease in water miscible organic solvents like, acetonitrile, dimethylformamide (DMF), ethanol, methanol, and propanol is described. The organic solvents exhibited a concentration dependent inhibitory effect on both the immobilized and the soluble urease in the presence of urea. Pretreatment of soluble enzyme preparations with organic solvents in the absence of substrate for 10 min at $30^{\circ}C$ led to rapid loss in the activity, while similar pretreatment of immobilized urease with 50% (v/v) of ethanol, propanol, and acetonitrile was ineffective. Time-dependent inactivation of immobilized urease, both in the presence and in the absence of urea, revealed stability for longer duration of time even at very high concentration of organic solvents. The soluble enzyme, on the other hand, was rapidly inactivated even at fairly lower concentrations. The results suggest that the immobilization of watermelon urease in calcium alginate make it suitable for its application in organic media. The observations are discussed.

• Korean Journal of Soil Science and Fertilizer
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• v.17 no.1
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• pp.77-81
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• 1984

Paddy soil samples were collected from the plow layers of 19 soil series distributed in Yeongnam district to determine the relationship between soil urease activity and other soil characteristics. The results obtained were summarized as follows: 1. Simple correlation analyses showed that the urease activity was positively related with available $P_2O_5(r=.844^{**})$, potassium activity ratio($r=0.762^{**}$), available $SiO_2(r=.580^{**})$ and $SiO_2$/O.M ratio($r=0.591^{**}$). 2. Among soil chemical properties which had positive linear correlations with soil urease activity, the content of available $P_2O_5$ in soil had the highest contribution to the multiple regression equation of soil urease activity. 3. The activity of soil urease was especially lower in sandy texture than in clayey paddy soils, and a tendency was observed that the heavier soil textures the higher activity of soil urease. 4. Relatively well drained soils had the higher activity of soil urease while the soils in "poorly drained" had remarkably lower activity of soil urease. 5. The soils in higher classes of paddy soil equitability group had higher activities of soil urease.

• Microbiology and Biotechnology Letters
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• v.20 no.5
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• pp.519-526
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• 1992

The gene coding for urease of alkalophilic Bacillus pasteurii had been cloned in Escherichia coli previously. The urease protein was purified 63.1-fold by TEAE-cellulose, DEAE-Sephadex A-50, Sephadex G-150 and Sephadex G-200 chromatographies with a 7.3% yield from the sonicated fluid of the E. coli HB1Ol(pBUll) encoding B. pasteurii urease gene. The ureases of E. coli (pBUll) and B. pasteurii possessed as a $K_m$ for urea, 42.1 mM and 40.4 mM, respectively. They hydrolyzed urea with $V_{max}$ of 86.9$\mu$mol/min and 160$\mu$mol/min, respectively. Both ureases were composed with four subunits (Mrs 67,000) and a subunit (Mr 20,000). The molecular weight of both native enzymes was Mr 280,OOO$pm$10,000 determined by gel filtration chromatography and Coomassie blue staining of the subunits. The optimal reaction pH of both ureases were pH 7.5. The ureases were stabled in pH 5.5-10.5. The optimal reaction temperature of both ureases were $60^{\circ}C$, and the ureases were stable for an hour at $50^{\circ}C$, 40min at $60^{\circ}C$ and 10 min at $70^{\circ}C$ The activity of both enzymes were inhibited completely by $Ag^{2+}$, $Hg^{2+}$, $Zn^{2+}$, $Cu^{2+}$, and were inhibited 60% by CoH, 30% by $Fe^{2+}$ and 10% by $Pb^{2+}$. However it was increased by the addition of $Sn^{2+}$, $Mn^{2+}$, $Mg^{2+}$ at concentration of $1{\times}10^{-3}$M. Both ureases were inhibited completely by p-CMB and acetohydroxamic acid. The urease expressed in E. coli (pBU11) was inhibited 70% by SDS. The urease of B. pasteurii was inhibited 40% by hydroxyurea, whereas the recombinant urease of E. coli strain was inhibited 17%. Both enzymes were not inhibited by cyclohexanediaminetetraacetic acid (CDTA) and ethylendiaminetetraacetic acid (EDTA).

• The Journal of the Korean Society for Microbiology
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• v.34 no.6
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• pp.533-542
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• 1999

H. pylori produces urease abundantly amounting to 6% of total protein of bacterial mass. Urease genes are composed of a cluster of 9 genes of ureC, ureD, ureA, ureB, ureI, ureE, ureF, ureG, ureH. Production of H. pylori urease in E. coli was studied with genetic cotransformation. Structural genes ureA and ureB produce urease apoprotein in E. coli but the apoprotein has no enzymatic activity. ureC and ureD do not affect urease production nor enzyme activity ureF, ureG, and ureH are essential to produce the catalytically active H. pylori urease of structural genes (ureA and ureB) in E.coli. The kinetics of activation of H. pylori urease apoprotein were examined to understand the production of active H. pylori urease. Activation of H. pylori urease apoprotein, pH dependency, reversibility of $CO_2$ binding, irreversibility of $CO_2$ and $Ni^{2+}$ incorporation, and $CO_2$ dependency of initial rate of urease activity have been observed in vitro. The intrinsic reactivity (ko) for carbamylation of urease apoprotein co expressed with accessory genes was 17-fold greater than that of urease apoprotein expressed without accessory genes. It is concluded that accessory genes function in maximizing the carbamylating deprotonated ${\varepsilon}$-amino group of Lys 219 of urease B subunit and metallocenter of urease apoprotein is supposed to be assembled by reaction of a deprotonated protein side chain with an activating $CO_2$ molecule to generate ligands that facilitate productive nickel binding.

• Microbiology and Biotechnology Letters
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• v.45 no.3
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• pp.265-270
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• 2017

A key virulence factor for urinary tract pathogens is the enzyme urease, which catalyzes the hydrolysis of urea into ammonium ions and carbonic acid. Urease activity plays an important role in the pathogenesis of urinary tract infection. In this study, the inhibitory effect of zerumbone against six urease-producing bacteria (Klebsiella oxytoca, K. pneumoniae, Morganella morganii, Proteus mirabilis, P. vulgaris, and Staphylococcus saprophyticus) and their urease activities were evaluated. The results of the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) tests showed that zerumbone had antibacterial effect against these six urease-producing bacteria. The MIC and MBC of zerumbone ranged from 0.5 to 2 mM and 1 to 4 mM, respectively. In the urease inhibitory assay, zerumbone showed better urease inhibition ($56.28{\pm}2.45-37.83{\pm}3.47%$) than the standard urease inhibitor, acetohydroxamic acid ($40.46{\pm}1.94-22.99{\pm}3.53%$). However, zerumbone did not affect the levels of the urease subunit. These results clearly indicated that zerumbone has antibacterial potential against urease-producing bacteria and possesses excellent bacterial urease inhibition properties.

• Journal of Microbiology and Biotechnology
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• v.10 no.6
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• pp.865-872
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• 2000

Helocobacter phylori is the major cause of gastritis, peptic ulcer, and a principal risk factor for gastric cancer. As the firs step towards a vaccine against H. pylori infection, Hy.pylori urease was expressed and purified as a recombinant apoenzyme (rUrease) in E. coli. In order to develop an effective immunization protocol using rUrease, the host immune responses were evaluated after the oral immunization of mice with rUrease preparations plus cholera toxin relative to various conditions, such as the physical nature of the antigen, the frequency of the booster immunization, the dose of the antigen, and the route of administration. The protective efficacy was assessed using a quantitative culture following an H. pylori SS1 challenge. It was demonstrated that rUrease, due to its particulated nature, was more superior than the UreB subunit as a vaccine antigen. The oral immunization of rUrease elicited significant systemic and secretory antibody responses, and activated predominantly Th2-type cellular responses. The bacterial colonization was significantly reduced (~100-fold) in those mice immunized with three or four weekly oran doses of rUrease plus cholera toxin (p<0.05), when compared to the non-immunized/challenged controls. The protection correlated well with the elicited secretory IgA level against rUrease, and these secretory antibody responses were highly dependent on the frequency of the booster immunization, yet unaffected by the dose of the antigen (25-200$\mu\textrm{g}$). These results demonstrate the remarkable potential of rUrease as a vaccine antigen, thereby strengthening the possibility of developing an H. pylori vaccine for humans.

• Journal of Microbiology and Biotechnology
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• v.4 no.2
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• pp.108-112
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• 1994

The genetic organization of the urease gene cluster from an alkalophilic Bacillus pasteurii was determined by subcloning and Tn5 transposon mutagenesis of a 10.7 kilobasepair cloned fragment. A region of DNA between 5.0 and 6.0 kb in length is necessary for urease activity. In vitro transcription-translation analysis of transposon insertion mutants of the cloned urease genes demonstrated that the major ($M_r$ 67,000) and minor ($M_r$ 20,000) structural peptides of urease are encoded at one end of the urease gene cluster and at least 3 additional polypeptides are encoded by adjacent DNA sequences.