• 상하수도학회지
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• 제32권5호
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• pp.399-409
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• 2018

Prussian blue is known as a superior material for selective adsorption of radioactive cesium ions; however, the separation of Prussian blue from aqueous suspension, due to particle size of around several tens of nanometers, is a hurdle that must be overcome. Therefore, this study aims to develop granule type adsorbent material containing Prussian blue in order to selectively adsorb and remove radioactive cesium in water. The surface of granular activated carbon was grafted using a covalent organic polymer (COP-19) in order to enhance Prussian blue immobilization. To maximize the degree of immobilization and minimize subsequent detachment of Prussian blue, several immobilization pathways were evaluated. As a result, the highest cesium adsorption performance was achieved when Prussian blue was synthesized in-situ without solid-liquid separation step during synthesis. The sample obtained under optimal conditions was further analyzed by scanning electron microscope-energy dispersive spectrometry, and it was confirmed that Prussian blue, which is about 9.7% of the total weight, was fixed on the surface of the activated carbon; this level of fixing represented a two-fold improvement compared to before COP-19 modification. In addition, an elution test was carried out to evaluate the stability of Prussian blue. Leaching of Prussian blue and cesium decreased by 1/2 and 1/3, respectively, compared to those levels before modification, showing increased stability due to COP-19 grafting. The Prussian blue based adsorbent material developed in this study is expected to be useful as a decontamination material to mitigate the release of radioactive materials.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2001년도 추계학술발표대회
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• pp.671-674
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• 2001

정제, 농축된 free CGTase 는 pH 6.0 - 7.0의 범위에서 안정하였으며, $70^{\circ}C$정도의 고온에서도 안정한 특성을 가지고 있었으며, CGTase의 고정화를 위한 방법으로서 CNBr-activated sepharose 4B에 의한 고정화가 가장 높은 효율올 나타내는 것으로 결정되었다. 고정화의 최적조건은 $30^{\circ}C$, 60rpm, pH 6.0의 phosphate buffer 하에서 9 시간 동안 고정화하는 것이었고, CNBr-activated sepharose 4B를 이용한 고정화의 경우 실험에 사용된 다른 모든 이온교환 수지에서 얻어진 효율에 비해 월등한 효과를 보였다.

• Journal of Applied Biological Chemistry
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• 제66권
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• pp.512-518
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• 2023

To address inherent weaknesses such as low mechanical strength and limited enzyme loading capacity in conventional chitosan or alginate beads, an additional step involving the exchange of anionic surfactants with hydroxide ions was employed to prepare porous chitosan hydrogel capsules for enzyme immobilization. Consequently, excellent thermal stability and long-term storage stability were confirmed. Furthermore, coating the porous chitosan hydrogel capsules with polydopamine not only improved mechanical stability but also exhibited remarkable enzyme immobilization efficiency (97.6% for M1-D0.5). Additionally, it was demonstrated that the scope of application for chitosan hydrogel beads, prepared using conventional methods, could be further expanded by introducing an additional step of polydopamine coating. The enzyme immobilization matrix developed in this study can be selectively applied to suit specific purposes and is expected to be utilized as a support for the adsorption or covalent binding of various substances.

• Journal of Microbiology and Biotechnology
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• 제17권4호
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• pp.650-654
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• 2007

In our previous work, a method of pretreating lipase was developed to prevent loss of its activity during covalent immobilization. In this study, Rhizopus oryzae lipase was pretreated before immobilization and then immobilized on a silica gel surface. The effects of the various materials and conditions used in the pretreatment stage on the activity of immobilized lipase were investigated. Immobilized lipase pretreated with 0.1% of soybean oil had better activity than those pretreated with other materials. The optimal temperature, agitation speed, and pretreating time for lipase pretreatment were determined to be $40^{\circ}C$, 200rpm, and 45min, respectively. The activity of immobilized soybean oil pretreated lipase was 630U/g matrix, which is 20 times higher than that of immobilized non-pretreated lipase. In addition, immobilized lipase activity was maintained at levels exceeding 90% of its original activity after 10 reuses.

• Journal of Microbiology and Biotechnology
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• 제26권5호
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• pp.829-836
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• 2016

Penicillin G acylase (PGA) was immobilized on magnetic Fe₃O₄@chitosan nanoparticles through the Schiff base reaction. The immobilization conditions were optimized as follows: enzyme/support 8.8 mg/g, pH 6.0, time 40 min, and temperature 25 ℃. Under these conditions, a high immobilization efficiency of 75% and a protein loading of 6.2 mg/g-support were obtained. Broader working pH and higher thermostability were achieved by the immobilization. In addition, the immobilized PGA retained 75% initial activity after ten cycles. Kinetic parameters V_max and K_m of the free and immobilized PGAs were determined as 0.113 mmol/min/mg-protein and 0.059 mmol/min/mg-protein, and 0.68 mM and 1.19 mM, respectively. Synthesis of amoxicillin with the immobilized PGA was carried out in 40% ethylene glycol at 25 ℃ and a conversion of 72% was obtained. These results showed that the immobilization of PGA onto magnetic chitosan nanoparticles is an efficient and simple way for preparation of stable PGA.

• Biotechnology and Bioprocess Engineering:BBE
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• 제9권6호
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• pp.465-470
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• 2004

Optimal conditions for the in situ immobilization of lipase in aldehyde-silica packed columns, via reductive amination, were investigated. A reactant mixture, containing lipase and sodium borohydride (NaCBH), was recirculated through an aldehyde-silica packed column, such that the covalent bonding of the lipase, via amination between the amine group of the enzyme and the aldehyde terminal of the silica, and the reduction of the resulting imine group by NaCBH, could occur inside the bed, in situ. Mobile phase conditions in the ranges of pH $7.0{\~}7.8$, temperatures between $22{\~}28^{circ}C$ and flow rates from $0.8{\~}1.5\;BV/min$ were found to be optimal for the in situ immobilization, which routinely resulted in an immobilization of more than 70 mg­lipase/g-silica. Also, the optimal ratio and concentration for feed reactants in the in situ immobilization: mass ratio [NaCBH]/[lipase] of 0.3, at NaCBH and lipase concentrations of 0.75 and 2.5 g/L, respectively, were found to display the best immobilization characteristics for concentrations of up to 80 mg-lipase/g-silica, which was more than a 2-fold increase in immobilization compared to that obtained by batch immobilization. For tributyrin hydrolysis, the in situ immobilized lipase displayed lower activity per unit mass of enzyme than the batch-immobilized or free lipase, while allowing more than a $45\%$ increase in lipase activity per unit mass of silica compared to batch immobilization, because the quantity of the immobilization on silica was aug­mented by the in situ immobilization methodology used in this study.

• KSBB Journal
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• 제21권4호
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• pp.279-285
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• 2006

본 연구에서는 trypsin을 모델 단백질로 하여 단백질 본연의 환성을 유지할 수 있는 고정화 방법을 찾기 위하여 공유결합방법과 친화력 결합방법을 이용하여 trypsin을 고정화 하였다. Streptavidin-biotin system을 이용한 고정화 방법은 bioactivity 유지측면에서 공유결합 방법보다 우수함을 확인하였다. 하지만 streptavidin-biotin system을 이용하였을 때 고정화 수율이 낮은 것은 해결해야 할 과제이다. 분자량이 다른 기질들(BAPNA, insulin, BSA)을 대상으로 고정화 trypsin의 부위 특이적 절단 특성을 분석한 결과 streptavidin-biotin에 의해 고정화된 trypsin이 절단효율도 높고 sequence coverage도 높은 것으로 확인되었다. 또한 공유결합된 trypsin은 견고한 분자구조를 나타낸 반면 streptavidin-biotin system으로 고정화된 trypsin은 유연성이 높은 것을 QCM-D를 이용하여 관찰할 수 있었다. 따라서 streptavidin-biotin system에 의한 고정화 방법에서 streptavidin-biotin 결합이 일종의 spacer arm 역할을 하면서 고정화된 trypsin의 분자유연성을 향상시켜 절단반응의 부위특이성과 절단수율을 향상시키는 것으로 판단되었다.

• KSBB Journal
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• 제8권4호
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• pp.320-327
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• 1993

스티렌/아크렬 라텍스 폴리머를 폴리에릴렌이민으 로 표면처 리하여 얻은 PEI-microspheres는 그 표면 화학적 특성이 효소의 고정화에 적합하여 배당체의 합성반응에 서 필요로 하는 alginate-enclosed micro­s spheres biocatalyst를 제조할 수 있었고 라텍스 폴리머 표변에 형성된 폴리에틸렌이민 피막은 효소 부근에 친수성 분위기를 유지하여 높은 효소활성을 나 타내었을 뿐만 아니라, 생성된 배당체가 효소에 접 근하여 가수분해되는 반응을 억제하여 배당체의 수 율과 농도를 크게 증가시 켰으므로 alginate-en closed microspheres에 의한 배당체의 합성반응이 연속생산공정으로 연구개발 될 수 있다.

• Biotechnology and Bioprocess Engineering:BBE
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• 제8권4호
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• pp.263-267
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• 2003

Magnetic nanoparticles prepared from an alkaline solution of divalent and trivalent iron ions could covalently bind protein via the activation of Nethyl-N-(3-dimethylaminopropyl) carbodiimide (EDC). Trypsin and avidin were taken as the model proteins for the formation of protein-nanoparticle conjugates. The immobilized yield of protein increased with molar ratio of EDC/nanoparticie. Higher concentrations of added protein could yield higher immobilized protein densities on the particles. In contrast to EDC, the yields of protein immobilization via the a ctivation of cyanamide were relatively lower. Nanoparticles bound with avidin could attach a single-stranded DNA through the avidin-biotin interaction and hybridize with a DNA probe. The DNA hybridization was confirmed by fluorescence microscopy observations. Immobilized DNA on nanoparticles by this technique may have widespread applicability to the detection of specific nucleic acid sequence and targeting of DNA to particular cells.

• 약학회지
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• 제42권1호
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• pp.53-58
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• 1998

A novel type of sulfotransferase, arylsulfate sulfotransferase (EC 2.8.2.22) purified from Haemophilus K-12, an intestinal bacterium of a mouse, was immobilized onto AH-S epharose 4B, CH-Sepharose 4B and DEAE-celluose. The enzyme was stabilized for storage more markedly by covalent immobilization onto AH-Spharose 4B or CH-Sepharose 4B and by adsorptive immobilization onto DEAE-cellulose than the free enzyme. The optimal pH and acceptor substrate specificity of immobuized enzyme were similar to those of the free enzyme.