• 제목/요약/키워드: Protein Immobilization

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Organic-Inorganic Hybrid Nanoflowers as Potent Materials for Biosensing and Biocatalytic Applications

  • Tran, Tai Duc;Kim, Moon Il
    • BioChip Journal
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    • 제12권4호
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    • pp.268-279
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    • 2018
  • Flower-shaped organic-inorganic hybrid nanostructures, termed nanoflowers, have received considerable recent attention as they possess greatly enhanced activity, stability, durability, and even selectivity of entrapped organic biomolecules, which are much better than those from the conventional methods. They can be synthesized simply via co-incubation of organic and inorganic components in aqueous buffer at room temperature and yield hierarchical nanostructures with large surface-to-volume ratios, allowing for low-cost production by easy scale-up, as well as the high loading capacity of biomolecules without severe mass transfer limitations. Since a pioneering study reported on hybrid nanoflowers prepared with protein and copper sulfate, many other organic and inorganic components, which endow nanoflowers with diverse functionalities, have been employed. Thanks to these features, they have been applied in a diverse range of areas, including biosensors and biocatalysis. To highlight the progress of research on organic-inorganic hybrid nanoflowers, this review discusses their synthetic methods and mechanisms, structural and biological characteristics, as well as recent representative applications. Current challenges and future directions toward the design and development of multi-functional nanoflowers for their widespread utilization in biotechnology are also discussed.

The GABAB receptor associates with regulators of G-protein signaling 4 protein in the mouse prefrontal cortex and hypothalamus

  • Kim, Gyeongwha;Jung, Soonwoong;Son, Hyeonwi;Kim, Sujeong;Choi, Jungil;Lee, Dong Hoon;Roh, Gu Seob;Kang, Sang Soo;Cho, Gyeong Jae;Choi, Wan Sung;Kim, Hyun Joon
    • BMB Reports
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    • 제47권6호
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    • pp.324-329
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    • 2014
  • Regulators of G-protein signaling (RGS) proteins regulate certain G-protein-coupled receptor (GPCR)-mediated signaling pathways. The GABAB receptor ($GABA_BR$) is a GPCR that plays a role in the stress response. Previous studies indicate that acute immobilization stress (AIS) decreases RGS4 in the prefrontal cortex (PFC) and hypothalamus (HY) and suggest the possibility of a signal complex composed of RGS4 and $GABA_BR$. Therefore, in the present study, we tested whether RGS4 associates with $GABA_BR$ in these brain regions. We found the co-localization of RGS4 and $GABA_BR$ subtypes in the PFC and HY using double immunohistochemistry and confirmed a direct association between $GABA_{B2}R$ and RGS4 proteins using co-immunoprecipitation. Furthermore, we found that AIS decreased the amount of RGS4 bound to $GABA_{B2}R$ and the number of double-positive cells. These results indicate that $GABA_BR$ forms a signal complex with RGS4 and suggests that RGS4 is a regulator of $GABA_BR$.

beta-Galactosidase의 고정화 및 응용에 관한 연구 제1보: Aspergillus niger CAD 1의 효소생산 조건 및 효소학적 성질 (Studies on immobilization and application of beta-galactosidase I. Conditions for production and properties of the enzyme from Aspergillus niger CAD 1)

  • 이용규;전순배;최원기;정기철;배석;김관천
    • 한국식품영양과학회지
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    • 제15권4호
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    • pp.32-39
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    • 1986
  • 토양에서 분리한 여러가지 곰팡이류 가운데 beta-galactosidase 생성력이 가장 좋은 Aspergillus niger CAD 1을 효소 생산 균주로 선정 하였다. 이 균주의 효소 생산 최적 배양조건은 밀기울에 0.5% 탈지 분유를 첨가한 배지를 $30{\circ}C$에서 72시간 정치배양하는 것이었다. 아세톤으로 침전시킨 조효소(crude enzyme)는 일차로 DEAE-cellulose, 2차로 Sephadex G-100 gel filtration에 의하여 1,387배로 정제되었고 이때의 수율은 6.2%이였다. 정제된 효소의 최적 온도는 $45{\circ}C$ 최적 pH는 4.5이었으며, 기질로서 ONPG와 유당에 대한 Km값은 각각 $3.57{\times}10^3M$$83.3{\times}10^3M$, Vmax값은 각각 33.0 unit/mg protein과 15.38unit/mg protein이었다. 활성화 에너지는 9,900cal/mol이었으며 효소활성 및 안정제로 금속이온을 필요로 하지 않았다. 50ml의 탈지유, 4.8%유당 용액, 유청용액에 효소 5ml(182 unit/ml)를 침가하여 $45^{\circ}C$에서 10시간 반응시켰을 때의 유당 분해율은 각각 65%, 70%, 78%를 나타내었다.

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우레아제(Urease) 고정막에 의한 요소(Urea)의 가수분해 (Hydrolysis of Urea by Immobilized Urease Membrane)

  • 김병식;김민;허광범;홍주희;나원재;김재훈
    • 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.

양극산화 임플란트 표면에 적용된 헤파린과 골형성단백질(rhBMP-2)이 치조골 증대에 미치는 효과: 방사선학적 평가 (Effect of immobilization of the recombinant human bone morphogenetic protein 2 (rhBMP-2) on anodized implants coated with heparin for improving alveolar ridge augmentation in beagle dogs: Radiographic observations)

  • 이소현;조재영;윤미정;전영찬;허중보;정창모
    • 대한치과보철학회지
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    • 제51권4호
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    • pp.307-314
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    • 2013
  • 연구 목적: 본 연구는 골형성단백질의 서방출을 위해 헤파린과 골형성단백질 (rhBMP-2)을 화학적으로 고정시킨 양극산화 티타늄 임플란트가 골 결손부에서 임플란트 주변의 수직적 골증대에 미치는 효과를 방사선학적으로 평가하고자 시행되었다. 연구 재료 및 방법: Pure-Titanium을 사용하여 길이 7.0 mm, 직경 3.5 mm의 실험용 임플란트 18개를 제작하였다. 모든 임플란트를 양극산화처리 하였고, 플랫폼 하방 2.5 mm에 식립 기준선을 표시하였다. rhBMP-2가 코팅되지 않은 임플란트 집단을 대조군으로, dip and dry 방법으로 rhBMP-2를 물리적 흡착시킨 집단을 BMP군, 3,4-dihydroxyphenylalanine(DOPA)-heparin을 이식하고 rhBMP-2를 화학적으로 고정시킨 집단을 Hep-BMP군으로 설정하였다. 각군별6개씩의 임플란트를 3마리의 비글견 양측 하악에 한쪽에 3개씩 총 18개를 치조정 상방으로 2.5 mm 노출시켜 식립하였다. 식립 직후와 4주, 8주에 식립부위의 방사선학적 검사가 시행되었고, 각 시기별, 각 군별 임플란트의 근원심 변연골의 수직적 재생량에 대한 평균값과 표준편차를 얻었다. Kruskal-Wallis test와 Mann-Whitney U test를 이용하여 4주, 8주에서 대조군과 실험군들의 차이를 비교 분석하고, 유의 수준5%에서 통계적으로 검정하였다. 결과:방사선학적 관찰 결과 임플란트 근원심 변연골 재생량(평균값 ${\pm}$ 표준편차)은 4주에 대조군은 $0.09{\pm}0.22mm$, BMP군은 $1.02{\pm}0.72mm$, Hep-BMP군은 $1.29{\pm}0.51mm$ 였으며, 8주에서는 각각 $0.11{\pm}1.26mm$, $1.11{\pm}0.58mm$, $1.59{\pm}0.79mm$였다. 두 실험군 모두 4주와 8주에서 대조군과 비교 시 유의한 수직적 골증대를 나타냈으나(P<.05), Hep-BMP군과BMP군의 비교에서는 유의한 변연골 재생량 차이를 보이지 않았다(P>.05). 결론: 골형성단백질을 물리적으로 흡착시키거나 서방출 위해 헤파린을 이용하여 화학적으로 고정시킨 양극산화 임플란트 표면은 모두 골 결손부에서 임플란트 주변골의 수직적 증대에 효과적이었다. 그러나 방사선학적 관찰의 한계 내에서 골형성단백질의 이 두가지 적용방법간에는 수직적 골증대량에 유의한 차이가 없었다.

Various Types and Manufacturing Techniques of Nano and Micro Capsules for Nanofood

  • Kim, Dong-Myong
    • Journal of Dairy Science and Biotechnology
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    • 제24권1호
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    • pp.53-63
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    • 2006
  • Nano and micro capsulation (NM capsulation) involve the incorporation for nanofood materials, enzymes, cells or other materials in small capsules. Since Kim D. M. (2001) showed that a new type of food called firstly the name of nanofood, which means nanotechnology for food, and the encapsulated materials can be protected from moisture, heat or other extreme conditions, thus enhancing their stability and maintaining viability applications for this nanofood technique have increased in the food. NM capsules for nanofood is also utilized to mask odours or tastes. Various techniques are employed to form the capsules, including spray drying, spray chilling or spray cooling, extrusion coating, fluidized bed coating, liposome entrapment, coacervation, inclusion complexation, centrifugal extrusion and rotational suspension separation. Each of these techniques is discussed in this review. A wide variety of nanofood is NM capsulated - flavouring agents, acids, bases, artificial sweeteners, colourants, preservatives, leavening agents, antioxidants, agents with undesirable flavours, odours and nutrients, among others. The use of NM capsulation for sweeteners such as aspartame and flavors in chewing gum is well known. Fats, starches, dextrins, alginates, protein and lipid materials can be employed as encapsulating materials. Various methods exist to release the ingredients from the capsules. Release can be site-specific, stage-specific or signaled by changes in pH, temperature, irradiation or osmotic shock. NM capsulation for the nanofood, the most common method is by solvent-activated release. The addition of water to dry beverages or cake mixes is an example. Liposomes have been applied in cheese-making, and its use in the preparation of nanofood emulsions such as spreads, margarine and mayonnaise is a developing area. Most recent developments include the NM capsulation for nanofood in the areas of controlled release, carrier materials, preparation methods and sweetener immobilization. New markets are being developed and current research is underway to reduce the high production costs and lack of food-grade materials.

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Dechlorination of High Concentrations of Tetrachloroethylene Using a Fixed-bed Reactor

  • Chang, Young-C.;Park, Chan-Koo;Jung, Kweon;Kikuchi, Shintaro
    • 한국환경보건학회지
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    • 제36권4호
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    • pp.323-336
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    • 2010
  • We evaluated the properties of a fixed-bed column reactor for high-concentration tetrachloroethylene (PCE) removal. The anaerobic bacterium Clostridium bifermentans DPH-1 was able to dechlorinate PCE to cis-1,2-dichloroethylene (cDCE) via trichloroethylene (TCE) at high rates in the monoculture biofilm of an upflow fixed-bed column reactor. The first-order reaction rate of C. bifermentans DPH-1 was relatively high at $0.006\;mg\;protein^{-1}{\cdot}l{\cdot}h^{-1}$, and comparable to rates obtained by others. When we gradually raised the influent PCE concentration from $30\;{\mu}M$ to $905\;{\mu}M$, the degree of PCE dechlorination rose to over 99% during the operation period of 2,000 h. In order to maintain efficiency of transformation of PCE in this reactor system, more than 6 h hydraulic retention time (HRT) is required. The maximum volumetric dechlorination rate of PCE was determined to be $1,100\;{\mu}mol{\cdot}d^{-1}l$ of reactor $volume^{-1}$, which is relatively high compared to rates reported previously. The results of this study indicate that the PCE removal performance of this fixed-bed reactor immobilized mono-culture is comparable to that of a fixed-bed reactor mixture culture system. Furthermore, our system has the major advantage of a rapid (5 days) start-up time for the reactor. The flow characteristics of this reactor are intermediate between those of the plug-flow and complete-mix systems. Biotransformation of PCE into innocuous compounds is desirable; however, unfortunately cDCE, which is itself toxic, was the main product of PCE dechlorination in this reactor system. In order to establish a system for complete detoxification of PCE, co-immobilization of C. bifermentans DPH-1 with other bacteria that degrade cDCE aerobically or anaerobically to ethene or ethane may be effective.

Surface Modification of Polypropylene Membrane by ${\gamma}$ Irradiation Methods and their Solutes Permeation Behaviors

  • Shim, J. K.;Lee, S. H.;Kwon, O. H.;Lee, Y. M.;Nho, Y. C.
    • 한국막학회:학술대회논문집
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    • 한국막학회 1998년도 춘계 총회 및 학술발표회
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    • pp.99-101
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    • 1998
  • 1. Introduction : The conventional grafting polymerization technique requires chemically reactive groups on the surface as well as on the polymer chains. For this reason, a series of prefunctionalization steps are necessary for covalent grafting. The surface prefunctionalizational technique for grafting can be used to ionization radiation, UV, plasma, ion beam or chemical initiators. Of these techniques, radiation method is one of the useful methods because of uniform and rapid creation of active radical sites without catalytic contamination in grafted samples. If the diffusion of monomer into polymer is large enough to come to the inside of polymer substrate, a homogeneous and uniform grafting reaction can be carried out throughout the whole polymer substrate. Radiation-induced grafting method may attach specific functional moieties to a polymeric substrate, such as preirradiation and simultaneous irradiation. The former is irradiated at backbone polymer in vacuum or nitrogen gas and air, and then subsequent monomer grafting by trapped or peroxy radicals, while the latter is irradiated at backbone polymer in the presence of the monomer. Therefore, radiation-induced polymerization can be used to modification of the chemical and physical properties of the polymeric materials and has attracted considerable interest because it imparts desirable properties such as blood compatibility. membrane quality, ion excahnge, dyeability, protein adsorption, and immobilization of bioactive materials. Synthesizing biocompatible materials by radiation method such as preirradiation or simultaneous irradiation has often used $\gamma$-rays to graft hydrophilic monomers onto hydrophobic polymer substrates. In this work, in attempt to produce surfaces that show low levels of anti-fouling of bovine serum albumin(BSA) solutions, hydroxyethyl methacrylate(HEMA) was grafted polypropylene membrane surfaces by preirradiation technique. The anti-fouling effect of the polypropylene membrane after grafting was examined by permeation BSA solution.

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Optimization of Culture Medium for Novel Cell-Associated Tannase Production from Bacillus massiliensis Using Response Surface Methodology

  • Belur, Prasanna D.;Goud, Rakesh;Goudar, Dinesh C.
    • Journal of Microbiology and Biotechnology
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    • 제22권2호
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    • pp.199-206
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    • 2012
  • Naturally immobilized tannase (tannin acyl hydrolase, E.C. 3.1.1.20) has many advantages, as it avoids the expensive and laborious operation of isolation, purification, and immobilization, plus it is highly stable in adverse pH and temperature. However, in the case of cell-associated enzymes, since the enzyme is associated with the biomass, separation of the pure biomass is necessary. However, tannic acid, a known inducer of tannase, forms insoluble complexes with media proteins, making it difficult to separate pure biomass. Therefore, this study optimizes the production of cell-associated tannase using a "protein-tannin complex" free media. An exploratory study was first conducted in shake-flasks to select the inducer, carbon source, and nitrogen sources. As a result it was found that gallic acid induces tannase synthesis, a tryptose broth gives higher biomass, and lactose supplementation is beneficial. The medium was then optimized using response surface methodology based on the full factorial central composite design in a 3 l bioreactor. A $2^3$ factorial design augmented by 7 axial points (${\alpha}$ = 1.682) and 2 replicates at the center point was implemented in 17 experiments. A mathematical model was also developed to show the effect of each medium component and their interactions on the production of cell-associated tannase. The validity of the proposed model was verified, and the optimized medium was shown to produce maximum cell-associated tannase activity of 9.65 U/l, which is 93.8% higher than the activity in the basal medium, after 12 h at pH 5.0, $30^{\circ}C$. The optimum medium consists of 38 g/l lactose, 50 g/l tryptose, and 2.8 g/l gallic acid.

고정화 글루코아밀라제의 성질과 응용 (Characteristics and Applications of Immobilized Glucoamylase)

  • 조성환;김재욱
    • Applied Biological Chemistry
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    • 제28권4호
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    • pp.233-238
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    • 1985
  • Glucoamylase를 $ZrO_2$로 피복된 96% porous glass에 azo-linkage를 형성시켜 결합하게 한후, 2.5% glutaraldehyde로 처리하여 효소를 고정화시켰다. 효소기질로는 용해도가 높고 점도가 낮은 30% enzyme thinned cornstarch (dextrose equivalent 값 : 24)를 사용하여 plug flow-column reactor에서 연속반응시켰다. 반응 최적 pH는 수용성효소의 5.0보다 alkaline 쪽으로 기울어져 7.0으로 나타났고, 고정화반응에 따라 열안정성이 높아지고 $40{\sim}60^{\circ}C$에서 최적 온도범위를 가리키며, Km값은 수용성 효소의 1.25mM보다 낮은 1.04mM값을 보여 주었다. 따라서, pH 7.0, $45^{\circ}C$에서 160시간 동안 corn starch를 기질로 효소반응을 시켜 glucose 90.3%, maltose 8.0%인 DE값 94.0인 전분당분해산물을 획득할 수 있었다.

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