• Advances in Traditional Medicine
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• 제5권1호
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• pp.43-47
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• 2005

Jack bean urease was immobilized on gelatin beads with the help of glutaraldehyde. The optimum immobilization (67.6%) was obtained at 30mg/ml gelatin concentration, 0.5 mg/bead enzyme protein concentration, 1 % glutaraldehyde and at $4^{\circ}C$ incubation temperature. The $t_{1/2}$ of immobilized urease was approximately 90 days at $4^{\circ}C$ compared with $t_{1/2}$ of 20 days for the soluble urease, under identical condition. The apparent optimum pH shifted from 7.3 to 8.0 when the urease was immobilized. The optimum stability temperature of immobilized urease was found to be $60^{\circ}C$ while that of soluble urease was $45^{\circ}C$. Time-dependent thermal inactivation studies showed monophasic kinetics for soluble urease and immobilized urease at $70^{\circ}C$, respectively. The immobilized urease beads stored at $4^{\circ}C$ showed practically no leaching over a period of 30 days. Here we are presenting an easy and economical way of immobilizing urease on the gelatin beads making it suitable for various applications.

• Biotechnology and Bioprocess Engineering:BBE
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• 제11권2호
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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.

• Journal of Pharmaceutical Investigation
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• 제15권3호
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• pp.93-99
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• 1985

For the effective immobilization of urease on chitosan matrix with glutaraldehyde, optimal activation methods were studied, and its enzymatic properties was investigated. In the stability of enzyme. the retained activity of the native urease was 55% after it was soaked af pH 7 for 10 hrs., while the retained activity of immobilized one was about 62% after soaked at pH 6.5-8.5 for the same time. After heat treatment at $60^{\circ}C$ for 10 hrs., the native urease lost the most of its activity, while immobilized urease retained 54% of its activity by the same treatment. The retained activity of immobilized urease did not decrease nearly when it was stored at room temperature for 25 days. From Linweaver-Burk plots, the $V_{max}$ value of native urease was $66{\mu}M/l$ and that of immobilized urease was $41{\mu}M/l$, while Km value 40mM/l for both enzymes was unaltered.

• 공업화학
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• 제18권1호
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• pp.10-16
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• 2007

본 연구에서는 인체의 해로운 요소를 분해하고 분해 생성물인 암모니아($NH_3$)의 제거를 위한 우레아제 고정막 제조와 가수분해 특성에 관하여 검토되었다. 우레아제 고정막은 방사선 그라프트 중합법에 의해 다공성 중공사막에 음이온 교환기로서 DEA를 도입한 후 요소의 가수분해를 위해 우레아제를 고정시켜 제조하였다. 음이온 교환기가 도입된 막에 우레아제가 고정된 경우 그라프트율이 증가할수록 고정량이 증가하였다. 이것은 그라프트율이 증가할수록 그라프트 체인이 신장하여 단백질이 다층으로 고정되기 때문이다. 한편, 가교제의 첨가는 우레아제 고정막의 세척 과정에서 발생하는 탈리 현상을 억제할뿐 아니라 막의 반복사용도 가능하게 하였다. 5 h의 가교 반응을 통하여 탈리현상이 발생되지 않는 우레아제 고정막을 제조하였으며, 이때 제조된 우레아제 고정막은 1 mol과 4 mol의 요소 용액에서 각각 98%와 50% 이상의 가수분해 성능을 나타내었다.

• Advances in Traditional Medicine
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• 제7권1호
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• pp.79-84
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• 2007

Urease was immobilized with calcium alginate by entrapment method in the form of spherical beads and stored in Tris/acetate buffer (pH 7.3) at $4^{\circ}C$. Urease immobilized at different concentration of alginate beads (3%, 4% and 5%) showed higher apparent $K_{m}$ values than the soluble urease. Furthermore, $K_{m}$ has been shown to be corelated with effective diffusion coefficient (De) at different concentration of alginate gel. The present study showed that diffusion and reaction contribute to control the overall rate.

• Bulletin of the Korean Chemical Society
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• 제23권8호
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• pp.1169-1172
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• 2002

• 한국막학회:학술대회논문집
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• 한국막학회 2004년도 추계 총회 및 학술발표회
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• pp.43-45
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• 2004

• 한국미생물학회:학술대회논문집
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• 한국미생물학회 2001년도 추계학술대회
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• pp.26-36
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• 2001

A novel approach of microbiologically-enhanced crack remediation (MECR) has been initiated and evaluated in this report. Under the laboratory conditions, Bacillus pasteurii was used to induce $CaCO_3$ precipitation as the microbial urease hydrolyzes urea to produce ammonia and carbon dioxide. The ammonia released in surroundings subsequently increases pH, leading to accumulation of insoluble $CaCO_3$. Scanning electron micrography (SEM) and x-ray diffraction (XRD) analyses evidenced the direct involvement of microorganisms in $CaCO_3$ precipitation. In biochemical studies, the primary roles of microorganisms and microbial urease were defined. Furthermore, the role of urease in $CaCO_3$ precipitation was characterized utilizing recombinant Escherichia coli that encoded B. pasteurii urease genes in a plasmid. Microorganisms immobilized in polyurethane (PU) polymer were applied to remediate concrete cracks. Although microbiologically- induced calcite precipitation enhanced neither the tensile strength nor the modulus of elasticity of the PU polymer, cement mortar whose crack was remediated with the cemaden polymer showed a significant increase in compressive strength. Through detailed investigation, MECR showed an excellent potential in cementing cracks in granite, concrete, and beyond.

• 한국막학회:학술대회논문집
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• 한국막학회 2004년도 춘계 총회 및 학술발표회
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• pp.49-52
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• 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)

• 대한전기학회:학술대회논문집
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• 대한전기학회 2003년도 하계학술대회 논문집 C
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• pp.1938-1940
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• 2003

본 연구는 요소 센서 제작을 위한 과정으로서, 전기화학적 방법을 이용한 다공질 실리콘 구조 형성과, PDV(Physical Vapor Deposition) 법에 의한 백금 박막 코팅 및 전기화학적 전도성 고분자 코팅과 urease 고정화 단계를 고찰하고 감도 특성을 제시 하였다. 전극 기질로서 B을 도우핑한 p-type 실리콘웨이퍼를 사용하였고, HF:$C_2H_5OH:H_2O$=1:2:1의 부피비를 갖는 에칭 용액에서 5분간 -7 $mA/cm^2$의 일정 전류를 가하여 폭 2 ${\mu}m$, 깊이 10 ${\mu}m$의 다공질 실리콘(PS) 충을 형성하였다. 그 위에 200 ${\AA}$의 Ti 층을 underlayer로서 증착하고, 2000 ${\AA}$의 Pt를 중착하여 PS/Pt 박막 전극을 제작하고, 전도성 고분자로서 polypyrrole (PPy), 또는 poly(3-mehylthiophene) (P3MT)을 전기화학적으로 코팅한 후, urease(EC 3.5.1.5, type III, Jack Bean, Sigma)를 고정화 하였다. 고정화 시 전해질 수용액의 pH는 7.4로 하여 urease표면이 음전하를 갖도록 하고, 전극에 0.6 V (vs. SCE(Saturated Calomel Electrode))의 일정 전압을 가함으로써 urease가 전도성 고분자 표면에 전기적으로 흡착되도록 하였다. 이상의 방법으로 제작한 요소 센서의 감도는 PPy와 P3MT를 전자 전달 매질로 사용한 경우, 각각 8.44 ${\mu}A/mM{\cdot}cm^2$와 1.55 ${\mu}A/mM{\cdot}cm^2$의 감도를 보였다.