• Title/Summary/Keyword: Immobilized media

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Recombinant Expression, Isotope Labeling and Purification of the Vitamin D Receptor Binding Peptide

  • Chae, Young-Kee;Singarapu, Kiran;Westler, W. Milo;Markley, John L.
    • Bulletin of the Korean Chemical Society
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    • v.32 no.12
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    • pp.4337-4340
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    • 2011
  • The vitamin D receptor binding peptide, VDRBP, was overexpressed as a fused form with the ubiquitin molecule in Rosetta(DE3)pLysS, a protein production strain of Escherichia coli harboring an induction controller plasmid. The fusion protein was bound to the immobilized metal ions, and the denaturation and renaturation of the fusion protein were performed as a part of the purification procedure. After the elution of the fusion protein, the peptide hormone was released from its fusion partner by using yeast ubiquitin hydrolase (YUH), and subsequently purified by reverse phase chromatography. The purity of the resulting peptide fragment was checked by MALDI-TOF mass and NMR spectroscopy. The final yields of the target peptide were around 5 and 2 mg per liter of LB and minimal media, respectively. The recombinant expression and purification of this peptide will enable structural and functional studies using multidimensional NMR spectroscopy and X-ray crystallography.

A Study on the Optimal Design in the Indirect Aerated Fluidized Bed (간접 포기식 유동상의 이상적 설계에 관한 연구)

  • 안송엽;김환홍;권희태
    • Journal of Environmental Science International
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    • v.8 no.1
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    • pp.95-100
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    • 1999
  • Process intensification without any increase in bed requires the exploitation of fluid mechanical phenomena as the basis for elegant solutions to the process engineering problems which result from the need to retain and control the immobilized biomass, and for biomass recovery. The fluidized bed biological reactor provides a solution to these needs. The wastewater treatment characteristics of the fluidized bed was filled with sand media. Indirect aeration were studied experimentally. The researcher was filled with sand particle size(0.60~0.42mm) in three reactors with different section area(A)/height(H), in the state BOD loading 4.5kg-$BOD_5/m^3$ㆍd, and under the fixed state of hydraulic retention time for around 32 minutes.

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Removal Characteristics of Toluene in Biofilters Packed with Reticulated-PU-Foams of Different Porosities (서로 다른 공극률의 망상형 폴리우레탄들이 충전된 바이오필터에서 톨루엔 제거 특성)

  • 명성운;남윤수;이용우;최호석
    • KSBB Journal
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    • v.18 no.6
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    • pp.448-454
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    • 2003
  • We studied on the removal of toluene vapors in a lab-scale biofilter. There are three biofilters packed with reticulated polyurethane foams of different porosities of 15, 25, 45 PPI (Pore Per Inch) as media. A toluene-degrading strain (Pseudomonas Putida KCCM 11348, ATCC 12633) was naturally immobilized on the filter media by circulating the culture media. Three biofilters were operated under different sets of continuous experiments, varying both the design and operation parameters such as the inlet toluene concentration and the flow rate. Maximum elimination capacity of 115.5g/㎥hr of biofilter packed with foams of 25 PPI was obtained for toluene degradation. The effect of operating conditions such as flow rate, inlet toluene concentration and porosity on the performance of the biofilter was investigated.

Enhancement of Cultivation Efficiency of Bifidobacterium longum Using Calcium Carbonate Buffer System (Calcium Carbonate Buffer System을 이용한 Bifidobacterium longum의 배양 효율 증대에 관한 연구)

  • Lee, Ki-Yong;Hwang, In-Bum;Heo, Tae-Ryeon
    • Korean Journal of Food Science and Technology
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    • v.29 no.1
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    • pp.126-132
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    • 1997
  • Calcium carbonate ($CaCO_3$) immobilized with alginate was studied as buffer system to enhance the cultivation efficiency of Bifidobacterium longum (ATCC 15707) which is inhibited at low pH. To test the bufferring effect of the immobilized $CaCO_3$ beads, pH value in each modified trypticase-proteose peptone-yeast (TPY) broth which is adjusted to pH 4.0 with acetic acid, lactic acid and complex solution of acetic and lactic acid, 3:2 (M:M) was tested by concentration of $CaCO_3$ bead and reaction time. The bufferring effect of $CaCO_3$ bead became higher with increasing the amount of $CaCO_3$ bead in the acidic solution. The growth rate of bifidobacteria and bufferring effect were examined in relation to the amount of $CaCO_3$ bead and concentration of glucose in the modified TPY media. The growth rate of bifidobacteria and bufferring effect were increased with increasing the amount of $CaCO_3$ bead and concentration of glucose. Also, the exponential time of bifidobacteria became longer with increasing the amount of $CaCO_3$ bead and concentration of glucose in the modified TPY media. When we observed the growth rate of bifidobacteria by the method of pH-controlled culture and $CaCO_3$ buffer system, the $CaCO_3$ buffer system was more effective than that of pH-controlled culture. Therefore, this $CaCO_3$ buffer system may be useful as a method to enhance of the cultivation efficiency of bifidobacteria.

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Removal of Ammonia and Nitrite in Water by Bacillus sp. A8-8 (Bacillus sp. A8-8에 의한 수질 중의 암모니아 및 아질산성 질소 제거)

  • 이용석;유주순;정수열;최용락
    • Journal of Life Science
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    • v.13 no.1
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    • pp.47-53
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    • 2003
  • The purpose of this study is to improve the system for biological nitrogen oxidizing process in sewage and wastewater. A bacterium having high abilities to oxidize of nitrogen was one of the possessed on Lab. The strain was identified to Bacillus sp. A8-8, based on the physiological and biochemical properties. And the strain has ability degradation crude oil. In comparison with oxidizing rates with changing initial pH and temperature, the strain Bacillus sp. A8-8 was nitrogen oxidizing ability and growth rate on the various of pH, temperature. oxidizing rates of the strain in sewage and wastewater were about 48% and 62%, respectively. The nitrogen oxidizing rate was increased in proportion to the initial concentration of glucose. The microorganism, Bacillus sp. A8-8, immobilized in ceramic carrier were evaluated for the oxidation of ammonia in culture media.

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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    • v.22 no.2
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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.

Ionic Recognition with Quinone-Derivatized Calixarenes in Solution and at Self-Assembled Monlayers

  • Kim Hasuck;Kang Sun Kil;Chung Taek Dong
    • Journal of the Korean Electrochemical Society
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    • v.3 no.2
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    • pp.69-71
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    • 2000
  • Redox-active calix[4]arenes with carboxylic acid and disulfide groups were prepared and spontaneous deposition on silver and gold surfaces was observed. Owing to their unusual structure, the calix[4]arenes exhibit selective affinity fur alkaline earth metal ions in aqueous media. When annular ionophores are immobilized on the surface, voltammetric and spectroscopic studies show the entrapment of metal ions. Furthermore, it was possible to reversibly capture and remove the ions using strong chelating agents such as ethylenediaminetetraacetic acid (EDTA).

A Study on Alkyl Glucoside Synthesis by Amphiphilic Phase Enzyme Reaction (양친매상 효소반응에 의한 알킬글루코시드의 합성연구)

  • 허주형;임교빈김해성
    • KSBB Journal
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    • v.11 no.5
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    • pp.511-517
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    • 1996
  • An amphiphilic phase enzyme reaction was used to synthesize alkyl glucosides from glucose and alkyl alcohol with immobilized ${\beta}$-glucosidase using four glycol ether cosolvents(monoglyme, diglyme, 2-methoxyethanol, and 1,4-dioxane). Monoglyme was shown to be the best cosolvent for the amphiphilic phase medium composed of water/cosolvent/alkyl alcohol admixture. To obtain high yield of alkyl glucoside by amphiphilic phase enzyme reaction, hydrophilicity-hydrophobicity of amphiphilic media and enzyme microenvironment was optimized from the viewpoints of substrate solubility, enzyme activity, water activity, and dynamic equilibrium between glucose alcoholysis and glucoside hydrolysis. Under optimum reaction conditions, the highest concentrations of hexyl, octyl, decyl, and dodecyl glucosides were 18.2, 9.68, 7.27, and 6.03g/L, respectively.

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Crystal Phase Changes of Zeolite in Immobilization of Waste LiCI Salt

  • KIM Jeong-Guk;LEE Jae-Hee;Lee Sung-Ho;KIM In-Tae;KIM Joon-Hyung;KIM Eung-Ho
    • Proceedings of the Korean Radioactive Waste Society Conference
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    • 2005.11b
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    • pp.176-181
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    • 2005
  • The electrolytic reduction process and the electrorefining process, which are being developed at the Korea Atomic Energy Research Institute (KAERI), are to generate molten waste salts such as LiCI salt and LiCI-KCI eutectic salt, respectively. Our goal in waste salt management is to minimize a total waste generation and fabricate a very low­leaching waste form such as a ceramic waste form. Zeolite has been known to one of the most desirable media to immobilize waste salt, which is water soluble and easily radiolyzed. Zeolite can be also used to the removal of fission products from the spent waste salt. Molten LiCI salt is mixed with zeolite A at $650^{\circ}C$ to form a salt-loaded zeolite, and then thermally treated in above $900^{\circ}C$ to become an immobilized product with crystal phase of $Li_{8}Cl_{2}$-Sodalite. In this work, a crystal phase changes of immobilization medium, zeolite, during immobilization of molten LiCI salt using zeolite A is introduced.

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Biochemical Application of IgG Fc-Binding Peptide: From Biochip to Targeted Nano Carrier

  • Chung, Sang J.
    • Proceedings of the Korean Vacuum Society Conference
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    • 2013.02a
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    • pp.110-111
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    • 2013
  • FcBP consisting of 13 amino acids specifically binds to Immunoglobulin G Fc domain. Initially, we utilized this peptide for preparation of antibody chip as a PEG composite for enhanced solubility. After then, the peptide conjugate was immobilized on agarose resin, resulting in highly efficient affinity column for antibody purification. The efficiency was comparable to commercial Protein A column. Recently, this peptide was conjugated with cell penetrating peptide (CPP) on a backbone of GFP, affording antibody transducer, which carries antibody into live cells by simple mixing of antibody and the transducer in cell culture media. Antibody transduction into cells was monitored by live cell imaging. More recently, the FcBP was fused to ferritin cage, which consists of 24 ferritin protein molecules. The FcBP-ferritin cage showed greatly increased binding affinity to human IgG. Its binding was analyzed by QCM and SPR analysis. Finally, it was selectively delivered by Herceptin to SKBR3, a breast cancer cell, over MCF10A, non-tumorigenic cells (Fig. 1). Fig. 1. Fluorescent microscopic images of SKBR3 breast cancer cells (A~C) and MCF10A breast cells (D~F) treated with Cy3-trastuzumab/fFcBP-Pf_Fn complexes. Trastuzumab and FcBP-Pf_Fn, which were labeled with Cy3 (Cy3-trastuzumab) and fluorescein (fFcBP-Pf_Fn), respectively, selectively targeted SKBR3 over MCF10A.

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