• Applied Chemistry for Engineering
• /
• v.18 no.1
• /
• pp.10-16
• /
• 2007

In this study, we examined the preparation and hydrolysis property of immobilized urease membrane to decompose harmful urea in the body and remove ammonia which was produced by its decomposition. Urease immobilized membrane was prepared by introducing anion-exchange group DEA into porous hollow-fiber membrane by radiation graft polymerization method, and immobilization of urease. When urease was immobilized at membrane introduced with anion-exchange group, the more increasing grafting rate, the more increasing immobilization amount. The result originates from the fact that a greater amount of protein was immobilized by forming a multilayer on the longer grafted chain. Meanwhile, the addition of the cross-linker was possible not only to suppress separation phenomenon produced during a washing process of immobilized urease membrane but also to enable the recycling of membrane. Urease Immobilized membrane with no separation phenomenon was prepared by cross-linking reaction for 5 h, and the hydrolysis rate of prepared urease immobilized membrane was over 98% and 50%, respectively, in 1 mol and 4 mol urea solutions.

• Proceedings of the Membrane Society of Korea Conference
• /
• 2004.11a
• /
• pp.43-45
• /
• 2004

• Journal of Microbiology and Biotechnology
• /
• v.11 no.2
• /
• pp.317-325
• /
• 2001

For heuristic purposes, the relative ratio of urease contents inside and outside cells was surveyed using nine ureB+ strains of Helicobacter pylori. the ratio of the enzyme specific activity appeared to vary greatly between the various H. pylori strains, ranging from 0.5 to 2.5. Besides the above compartment, urease was also richly found in the membrane fraction, especially in either peripheral or integral form. The urease distribution across the H. pylori membrane was significantly influenced by the ambient pH; the specific activity of external urease was highest at pH 5.5 with a narrow plateau, whereas the internal specific activity was highest within a pH range of 4.5 to 6.5 with a broad plateau. These finding strongly suggest that H. pylori urease is secretory and responded to the external pH. However, at pH 4.0 or below, no urease activity was detected in either the internal or external compartment, although an increase in the color development with 2,4,6-trinitrobenzene sulfonate (TNBS) was observed. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) demonstrated that these phenomena may be related to a specific proteolysis in certain proteins, including urease or ${\gamma}$-glutamyl transpeptidase. Interestingly, the effect of ammonium ions n alleviating the enzyme inactivation inside the H. pylori cells was remarkably similar to that of D-glucose. In addition, it would appear that the cation acted as a surrogate of not only $Na^+$ but also $K^+$ thereby increasing the H. pylori P-type ATPase activity. This is of great interest, as it implies that the urease action in H. pylori is indispensible at any locus.

• Proceedings of the Membrane Society of Korea Conference
• /
• 2004.05b
• /
• pp.49-52
• /
• 2004

생체 내에서의 요소 형성은 단백질이 아미노산으로 분해되어 인체에 남은 요소는 오줌으로 배출된다. 그러나 고농도의 urea의 경우 단백질을 변형시키게 된다[1-2]. 이러한 고 농도의 urea를 단백질 공정을 통해서 제거시키는 기술이 최근의 투석 과학이다. 그러나 이러한 방법은 urea의 제거와 함께 많은 양의 단백질과 양이온이 유출 및 오염의 문제가 많이 발생하고 있다[3].(중략)

• Journal of Life Science
• /
• v.18 no.2
• /
• pp.241-248
• /
• 2008

This study has been carried out to secretion antibodies for the purpose of preventing the infection of Helicobacter pylori and using them as a supplement for treatment. This experiments have been separated antigens from H. pylori and observed into antibody production and the agglutination of H. pylori for the separated antigens. As major antigenic proteins separated from H. pylori, the following could be verified: 12 kinds of band for whole cell (WC), seven kinds of band for outer membrane protein (OMP), three kinds of band for crude urease, and one kind of band for lipopolysaccharide (LPS). The IgG anti-H. pylori antibody of separated antigens showed $77.9{\pm}6.4{\mu}g/ml$ for we (L), $84.9{\pm}6.4{\mu}g/ml$ for OMP, and $123.8{\pm}2.9{\mu}g/ml$ for crude urease, at the same antigen concentration of $20{\mu}g/100ull$, which showed the most at the crude urease. And it turned out that the IgA antibodies were generated with $2.5{\pm}0.32{\mu}g/ml$ for WC (L), $2.0{\pm}0.43{\mu}g/ml$ for OMP, and $1.3{\pm}0.25{\mu}g/ml$ for crude urease, which demonstrated the most for WC (L) antigens. As a result of verifying the immunogenecity of antigenic protein through the Western blotting, major antigenic substances could be confirmed as follows: 10 kinds for WC, six kinds for OMP and three kinds for crude urease. The agglutination values on the H. pylori of the antibody were $2^5,\;2^5,\;2^6\;and\;2^7$ at the antigen serums of anti-WC (H), anti-WC (L), anti-OMP and anti-crude urease, respectively, which indicated the highest for the antigen serum of anti-crude urease. The urease activation-inhibiting absorbance of antigen serum created by each antigen was $0.14{\pm}0.01$ for WC (H), $0.16{\pm}0.01$ for WC (L), $0.18{\pm}0.03$ for OMP, and $0.18{\pm}0.04$ for urease, demonstrating a significant inhibiting effect, compared with $0.26{\pm}0.02$ of the control group.

• Journal of Microbiology and Biotechnology
• /
• v.13 no.5
• /
• pp.673-679
• /
• 2003

Glutamine and urea, abundant in body fluids or plasma, yield net ammonium ions upon hydrolysis by ${\gamma}-glutamyl$ transpeptidase (${\gamma}-GTP$) and urease, respectively, and these two enzymes are largely produced from Helicobacter pylori. To investigate bacterial potential of ammonium production, we first quantified those in whole-cell systems and found that the relative ratio of their amounts varied greatly, especially with pH values and the cell's aging. During the H. pylori cultivation, the ratio appeared to be inversely proportional to each other, showing a progressive increase of the ${\gamma}-GTP$ with decreasing of the urease. Under the urease-defective conditions due to low pH or coccoids, the bacterial cells still possessed a considerable amount of ${\gamma}-GTP$, which was found exclusively in the external compartment, therefore, the cell's ammonium production was found to be solely dependent upon glutamine, and the external ammonium concentration was constant without any contribution of urea concentration. Such ammonium constancy would definitely have an adverse effect on the host, because of its absolute requirement for vacuolar degeneration by H. pylori VacA, maximized at approximately 10 mM $NH_4Cl$. It was also found that, by using the metal-saturated membrane vesicles, ammonium ions were likely to be involved in the pH-dependent cation-flux across the H. pylori membrane, where the role of ${\gamma}-GTP$ in ammonium homeostasis around cells was suggested, especially under the hostile conditions against H. pylori.

• BMB Reports
• /
• v.30 no.5
• /
• pp.308-314
• /
• 1997

We investigated biotinylation of nitrocellulose membrane for immuno-filtration capture assay. In order to enhance the efficiency of biotinylation, nitrocellulose membranes were pretreated with several chemicals for the purpose of suitable protein absorption through surface modification. As a signal generating enzyme, urease was used and the concentration of avidin was optimized for the efficient binding kinetics between urease-biotin in liquid phase and biotinylated membrane in solid phase. For effective biotinylation, bovine serum albumin-biotin complexes could be immobilized at a concentration of $370\;{\mu}g$/stick ($4.4\;cm^2$). Among tested chemicals, polylysine (0.25%) showed a significant effect in biotinylation. Polylysine is thought to enhance surface area by extending unbound residues into solution. Time of treatment over 30 min and higher molecular weight of polylysines (58,100 dalton) showed positive effect on the enhancement of biotinylation. The result from this study may be useful for developing a new biosensor and other biofunctional membranes for examining molecular recognition.

• Microbiology and Biotechnology Letters
• /
• v.25 no.2
• /
• pp.129-136
• /
• 1997

Helicobacter pylori is a spiral-shaped, microaerophilic human gastric pathogen causing chronic-active gastritis in association with duodenal ulcer and gastric cancer. To investigate the possibility of H. pylori outer membrane proteins (OMPS) as the oral vaccine antigens, sarcosine-insoluble outer membrane fraction has been prepared from H. pylori NCTC 11637. The major OMPs having apparent molecular masses of 62 kDa, 54 kDa and 33 kDa were detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), which were identified as urease B subunit (UreB), heat shock protein (Hsp54 kDa) and urease A subunit (UreA), respectively. Minor protein bands of 57 kDa, 52 kDa, 40 kDa, 36 kDa and 31 kDa were also observed. The antigenicity of H. pylori OMPs and antigenic cross-reactivity among the strains were determined by immunoblot analysis using anti-H. pylori OMPs antisera or intestinal lavage solutions. The results showed that UreB, Hsp54 kDa, UreA and 40 kDa proteins vigorously stimulated mucosal immune response rather than systemic immunity. From this results, these proteins seemed to be useful as the antigen candidates for the oral vaccine. The immunoblotting results with surface proteins from eight isolated H. pylori strains were similar to that of H. pylori NCTC 11637. The IgA which had been arised from oral administration of H. pylori OMPs, was able to bind H. pylori whole-cells.

• Childhood Kidney Diseases
• /
• v.25 no.2
• /
• pp.112-116
• /
• 2021

Hyperammonemia is mainly caused by diseases related to liver failure. However, there are also non-hepatic causes of hyperammonemia, such as urinary tract infection (UTI) due to urease-producing organisms. Urease production by these bacteria induces a hydrolysis of urinary urea into ammonia that can cross the urothelial cell membrane and diffuse into blood vessels, leading to hyperammonemia. Delayed diagnosis and treatment of hyperammonemia can lead to lethal encephalopathy that can cause brain damage and life-threatening conditions. In the presence of obstructive uropathy, UTI by urease-producing bacteria can lead to more severe hyperammonemia due to enhanced resorption of ammonia into the systemic circulation. In this report, we present a case of acute severe hyperammonemic encephalopathy leading to brain death due to accumulation of ammonia in blood caused by Morganella morganii UTI in a 10-year-old girl with cloacal anomaly, causing obstructive uropathy even after multiple corrections.

• Journal of the Korean Chemical Society
• /
• v.36 no.3
• /
• pp.393-404
• /
• 1992

The response properties of continuous automated system using an enzyme reactor for determination of urea were investigated. The enzyme reactor was constructed to packed-bed form which filled with nylon-6 beads (42∼48 mesh), which immobilized urease with glutaraldehyde, in teflon tube (2 mm I.D., 20 cm length). The system was composed of the enzyme reactor, gas dialyzer, and tublar PVC-nonactin membrane ammonium ion-selective electrode as an indicator electrode in serial order. The response characteristics of this system were as follows. That is, the concentration range of linear response, slope of linear response, detection limit, and conversion percentage were $5.5{\times}10^{-6}$∼$2.4{\times}10^{-3}M$, 57.8 mV/decade, $1.5{\times}10^{-6}$, and 80.8%, respectively. The optimum buffer and life time of urease reactor were 0.01M Tris-HCl buffer solution (pH 7.0∼7.8) and 0.01M phosphate buffer solution (pH 6.9∼7.5) and about 150 days, respectively. And the urease reactor had no interferences of the other physiological materials.