• Title/Summary/Keyword: 효소 고정화

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Determination of Ethanol in Alcoholic Beverages by Alcohol Oxidase Sensor (Alcohol oxidase 효소센서를 이용한 알코올 음료 중의 에탄올 정량)

  • Lee, Ok-Kyung;Kim, Tai-Jin;Noh, Bong-Soo
    • Korean Journal of Food Science and Technology
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    • v.27 no.2
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    • pp.266-269
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    • 1995
  • In order to measure alcohol contents with speed and accuracy, alcohol sensor was prepared. Alcohol sensor was made by connecting with oxygen electrode after immobilized alcohol oxidase on nylon net with glutaraldehyde. Alcohol was determined by changing the rate of dissolved oxygen consumption using D.O. analyzer. Alcohol contents in alcoholic beverages were determined under the optimum conditions. The results were 0.71% in low-alcohol beverage, $4{\sim}5%$ in beers, 10.06% in wine, 16.12% in chungju, 25.71% in soju, and 6.18% in takju, respectively. The values by alcohol sensor showed an excellent correlation(r=0.999) with GC method.

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Carbon Dioxide Sequestration of Enzyme Covalently Immobilized on Porous Membrane (공유결합으로 다공성 막에 고정화된 효소에 의한 이산화탄소 포집)

  • Park, Jin-Won
    • KSBB Journal
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    • v.28 no.4
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    • pp.225-229
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    • 2013
  • Bovine Carbonic anhydrase (BCA) was immobilized on a submicro-porous membrane through covalent immobilization. The immobilization was conducted on the porous membrane surface with the treatment of polyethyleneimine, glutaraldehyde, and the anhydrase, in sequence. The immobilization was confirmed using X-ray photon spectrometer. The pH values of carbon-dioxide saturated solution with buffer were monitored with respect to time to calculate the catalytic activities of hydration of carbon-dioxide for free and immobilized CA. The catalytic rate constant values for free CA, immobilized CA on polystyrene nanoparticles, and immobilized CA on a porous cellulose acetate membrane were 0.79, 0.67, and 0.56 $s^{-1}$, respectively. Reusability was studied up to 10 cycles of $CO_2$ sequestration. The activity for the CA immobilized on the membrane was kept to 95% after 10 cycles, and comparable to the CA on the nanoparticles. The stabilities for heat and storage were also investigated for the three cases. The results suggested that the CA immobilized the membrane had the least loss rate of the activity compared to the others. From this study, the porous membrane was feasible as a carrier for the CA immobilization in hydration and sequestration of carbon-dioxide.

Biodiesel Production from Waste Cooking Oil Using Alkali Catalyst and Immobilized Enzyme 1. Fatty Acid Composition (알칼리 촉매와 고정화 효소를 이용한 폐식용유로 부터 바이오 디젤 생산 1. 지방산 조성)

  • Shin, Choon-Hwan
    • Journal of Environmental Science International
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    • v.19 no.10
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    • pp.1247-1256
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    • 2010
  • Since biodiesel as bioenergy is defined as ester compounds formed by esterification of animal/vegetable oils, in this study three vegetable cooking oils (market, waste and refined waste ones) were esterified by reactions of alkali catalyst and immobilized enzyme. The fatty acid composition of the formed ester compounds was analyzed to investigate the feasibility of biodiesel production. By lipolysis (i.e, hydrolysis of Triglyceride (TG)), all three vegetable oils used in this study were found to produce Diglyceride (DG), Monoglyceride (MD) and Fatty acid ethylester (FAEE). However, the amount of produced FAEE (which can be used as an energy source) was in the increasing order of market cooking oil, waste one and refined waste one. With NaOH catalyst, FAEE was produced about 24.92, 17.63 and 11.31 % for the respective oils while adding Lipozyme TL produced FAEE about 43.54, 38.16 and 24.47 %, respectively. This indicates that enzyme catalyst is more effective than alkali one for transesterification. In addition, it was found that the composition of fatty acids produced by hydrolysis of TG was unchanged with alkali and immobilized enzyme reactions. Thus it can be expected that stable conditions remain in the course of mixing with gasoline whose composition is similar to that of the fatty acids.

Development of Chemiluminescence Immunoassay(CIA) & ELISA for the Detection of Anti-sperm Antibodies in Male Serum (항정자 항체 검출을 위한 CIA 및 ELISA 개발을 위한 기초 연구)

  • Kim, S.C.;Lee, K.S.;Kim, Y.K.;Kim, C.K.;Choi, K.H.;Kwon, O.J.;Kim, J.B.
    • Clinical and Experimental Reproductive Medicine
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    • v.17 no.1
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    • pp.71-80
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    • 1990
  • New immunoassay systems for the detection of anti-sperm antibodies were developed. For this, sperm surface protein was purified by the immunoaffinity column prepared by the coupling of rabbit anti-human IgG antibodies to Sepharose-4B. Fraction eluted by tris-HCI buffer containing SDS showed a single band having molecular weight of about 60KD on electrophoresis. Enzyme HRP labelled goat anti-human IgG and chemiluminescence aminobutylethyl-isoluminol(ABEI) labelled rabbit anti-human IgG were used for ELISA and CIA, respectively. These two labelled conjugate bound well with human IgG. When serum dilution curves were made to titrate positive serums, two kinds of curves with steep and sluggish slopes were obtained Serum samples were categorized into 3 groups: positive, weak positive and negative based on slope of curve and O.D. values at 1:160 dilution of serum. When ELISA and CIA were compared to conventional method Kibrick test by the determinations of 62 male serums with different diagnosis, the results of ELISA and CIA agreed well, but both disagreed with that of Kibrick test. This study showed that purified sperm surface antigen can be used to develope solid-phase immunoassay systems such as ELISA and CIA which may eliminate the problems encounted the immobilization of living sperm in other tests.

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Detection of Heavy Metal Ions by the Cuvette Assay Measuring Urease Inhibitory Activity (Urease 저해활성 측정 cuvette assay에 의한 중금속 이온 검출)

  • Kim, Dong-Kyung;Park, Kyung-Rim;Kang, Eun-Mi;Park, In-Seon;Kim, Nam-Soo
    • Applied Biological Chemistry
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    • v.46 no.2
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    • pp.74-78
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    • 2003
  • To determine the urease inhibitory activity of various heavy metal ions, a photometric cuvette assay for measuring ammonia production was developed. In this assay, the absorbance values at 630 m were linearly increased according to the ammonia concentrations up to 3.0 mg/l (r : 0.998). The urease inhibitions upon addition of a single species of heavy metal ions were in the decreasing order of Hg(II) > Pb(II) > Cu(II) > Cd(II) > Zn(II) ions. As expected, the urease inhibitions at a fixed concentration of a single species and at varying concentrations of other species occurred in the additive way. The above results show the applicability of the current method to the selective detection on Hg(II) ions as well as the screening of heavy metal ions possibly present at various samples.

Enzyme-Linked Immuno-Sorbent Assay for Bovine Caseins (우 Casein의 면역효소분석법)

  • 염행철
    • Korean Journal of Animal Reproduction
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    • v.16 no.2
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    • pp.87-102
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    • 1992
  • A rapid, sensitive, and specific enzyme-linked immuno-sorbent assay (ELISA) for bovine casein was developed. Biotinylated casein and peroxidase-conjugated avidin were used in the assay with antibody separated from yolks of immunized hens. Caseins were biotinylated with sulfo-N-hydroxy succinimido biotin and peroxi-dase-conjugated avidin bound the biotinylated casein which became bound to immobilized anti-body on a microplate. The antibodies were specific for bovine $\alpha$- and $\beta$-caseins, and their cross-reactivities with whey proteins, IgG, and serum albumin from bovine were not detectable by ELISA and Western blot. Various sensitivities ranging from 2ng/ml to 20${\mu}\textrm{g}$/ml of casein were achieved, and were controlled by adding vanous concentrations of the biotinylated casein. Parallelism was observed between standard and sample curves. The coefficients of variation of intra-assays and inter-assays from the most sensitive assay were 5.5 and 5.7%, respectively, at the 50% displacement. Casein contents of peripaturient milk samples showed that casein secretion rapidly increased 3d prepartum.

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Isolation, Identification and Enzyme Properties of a Bacterium producing Alkaline Protease (Alkaline protease를 생산하는 미생물의 분리, 동정 및 효소성질)

  • Shin, Kong-Sik;Kang, Sang-Mo;Ko, Jung-Youn
    • Applied Biological Chemistry
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    • v.43 no.3
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    • pp.169-173
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    • 2000
  • For the development of enzyme detergent capable of effectively washing at low temperature, a bacterium producing alkaline protease was isolated from soil samples, and properties of the enzyme were investigated. The selected strain was Gram negative, rod shape$(0.6{\sim}0.7{\times}1.3{\sim}2.6\;{\mu}m\;in\;size)$ and motile. It had the degradation activity of aesculin, gelatin and casein, and was catalase-positive. The cell wall components was meso-DAP, and G+C mole contents was 43.3%. From these results, the strain was identified as Acinetobacter sp. KN-27. The activity of alkaline protease by this strain peaked with 3,300 D.U/mL after 36 hours in the liquid culture at $40^{\circ}C$. The optimal pH and temperature of the enzyme were pH 9 and $60^{\circ}C$, respectively. Alkaline protease produced by Acinetobacter sp. KN-27 has shown two active bands on the electrophoresis of native gel.

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경유의 Model solution에서 고정화효소를 이용한 Dibezothiophene의 산화

  • Heo, Jeong-Chan;Seong, Hyeon-Tae;Ryu, Geun-Gap
    • 한국생물공학회:학술대회논문집
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    • 2001.11a
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    • pp.517-520
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    • 2001
  • Fossil fuels such as coal and crude oil contain various organic sulfur compounds. Combustion of these fuels emit sulfur oxides which are considered as msjor air pollutants causing acid rain problem. Among various organic sulfur compounds, aromatic sulfur compounds of thiophenes which constitute major sulfur fractions of heavy oils are not easily removed by hydrodesulfurization. Many peroxidase and hemoproteins are known to oxidize dibenzothiophene (DBT) to dibenzothiophene-sulfoxide(DBT - sulfoxide) then dibenzothiophene- sulfone (DBT-sulfone). The oxidation of DBT by the immobilized hemoproteins in n-octane was increased significantly when the hemoproteins were deposited on celites of the particle size between 0.75 - 1.0 mm and a conventional substrates. such as t-butyl hydroperoxide and cumene hydroperoxide. In anhydrous organic solvents with log P values larger than 4.0 DBT was completely oxidized by cumene hydroperoxide catalyzed by cytochrome c deposited on celites.

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The Effect of Chitosan on Elctrospinning of Silk Fibroin (실크 피브로인의 전기방사 거동에 미치는 키토산의 영향)

  • Yu, Su-Yeon;Jeong, Lim;Park, Won-Ho
    • Proceedings of the Korean Fiber Society Conference
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    • 2003.10b
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    • pp.267-268
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    • 2003
  • 실크 피브로인은 천연 고분자 물질 중 하나로 Bombyx mori와 Antheraea perni 등이 있으며 생체적 합성을 가지고 있어 효소고정화를 위한 매트릭스나 세포배양용 지지체와 같은 의료용 분야에서 활발히 연구되고 있다. 또한 키토산은 키틴이 50% 이상 탈아세틸화된 것으로 갑각류, 곤충류, 균류 등에 존재하며 창상피복재, 인조 피부 등의 의료 분야에서 광범위하게 사용되고 있다. 본 실험에서는 실크 피브로인에 단독으로는 전기방사가 되지 않는 키토산을 다양한 비율로 블렌드하여 키토산이 실크 피브로인의 전기방사된 섬유의 형태에 미치는 영향에 대하여 알아보았다. (중략)

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Synthesis of Osmium Redox Complex and Its Application for Biosensor Using an Electrochemical Method (오스뮴 착물 합성과 전기화학적인 방법을 이용한 바이오센서에 관한 연구)

  • Choi, Young-Bong;Kim, Hyug-Han
    • Journal of the Korean Electrochemical Society
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    • v.10 no.2
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    • pp.150-154
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    • 2007
  • Redox complexes to transport electrodes from biomaterial to electrodes are very important part in commercial biosensor industry. A novel osmium redox complex was synthesized by the coordinating pyridine group with osmium metal. A novel osmium complex is described as $[Os(dme-bpy)_2(ap-im)Cl]^{+/2+}$. We have been studied the electrochemical characteristics of this osmium complex with electrochemical techniques such as cyclic voltammetry and chronoamperommetry. In order to immobilize osmium redox complexes on the electrode, we deposited gold nano-particles on screen printed carbon electrode(SPE). The electrical signal converts the osmium redox films into an electrocatalyst for glucose oxidation. The catalytic currents were monitored that the catalytic currents were linearly increased from 1 mM to 5 mM concentrations of glucose.