• Title/Summary/Keyword: immobilized

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Development of an Immobilized Adsorbent for in situ Removal of Ammonium Ion from Mammalian Cell Culture Media and its Application to a Mammalian Cell Bioreactor: I. Development of Immobilized Adsorbent System (동물세포 배양액으로부터 암모늄 이온의 동시제거를 위한 고정화 흡착제의 개발과 동물세포 배양 시스템에의 응용: I. 고정화 흡착시스템 개발)

  • 박병곤;민용원;전계택;김익환;정연호
    • KSBB Journal
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    • v.13 no.4
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    • pp.404-410
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    • 1998
  • Three types of adsorbents were developed by immobilizing synthetic zeolite, Philipsite-Gismonine, in alginate, cellulose acetate and dialysis membrane for the in situ removal of ammonium ion which inhibits growth and productivity of animal cells such as CHO cells producing tPA. Ammonium ion removal efficiency and cell growth promoting effect with various immobilized adsorbents were evaluated and the membrane type was selected as an optimal immobilized adsorbent. The experiments were then simulated by adding 8mM ammonium chloride and immobilized adsorbent in order to validate the removal effect under high density cell cultures. The results showed increase in maximum cell density by three times, in cell viability, and in tPA productivity by 40%. And it was found that the promoting effects were more significant in case of high ammonium ion concentration system. It was also found that the optimum addition time for immobilized adsorbents was 48 hr in the absence of ammonium chloride addition and 72 hr in the presence of ammonium chloride addition.

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Degradation of Phenol by Activated Sludge Immobilized with Photo-crosslinked Resin (광경화성 수지에 고정화된 활성슬러지에 의한 페놀 분해)

  • 김선일;윤영재정경훈
    • KSBB Journal
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    • v.11 no.5
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    • pp.577-585
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    • 1996
  • Effects of various factors on the phenol degradation by activated sludge immobilized with the photo-crosslinked resin were investigated. The optimum pH on the degradation of phenol in both free and immobilized activated sludge was 7. When the pH of the reaction was varied from 5 to 10, the relative activity of the phenol degradation by the immobilized activated sludge was higher than that by the free activated sludge. A higher rate of phenol degradation was observed when a bead size was smaller. The phenol degradation in the free activated sludge was inhibited at the 3000 mg/L of phenol, while that in the immobilized activated sludge was maintained at the same concentration for 28 hrs without an inhibition. The degradation rates of phenol were not directly proportional to the increasing amount of immobilized beads dosage, but the phenol degradation was made in a rather short time than that for a free sludge system. The relative activities of the immobilized activated sludge after 7 runs of repeated reactions increased about 8 times as that of the first reaction. The activities for the phenol degradation remained stable for at least 80 days when the immobilized activated sludge was stored at an aerobic condition in the wastewater containing phenol. The loading rate as high as 5.59 kg-pheno1/㎥.d could have been achieved during the continuous treatment of phenol by the immobilized activated sludge.

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Studies on the Formation of Pyridoxal Phosphate by Immobilized Cells (고정화 균체에 의한 Pyridoxl Phosphate의 생산에 관한 연구)

  • Chu, Young-Ha;Tani, Yoshiki;Lee, Taik-Soo;Yu, Tai-Jong
    • Korean Journal of Food Science and Technology
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    • v.9 no.3
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    • pp.183-189
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    • 1977
  • Studies were made of the continuous production of Pyridoxal 5'-phosphate (pyridoxal-p) on simultaneously immobilized cell column. Whole-cell of Pseudomonas polycolor having high activity of pyridoxine 5'-phosphate (pyridoxine-p) oxidase and Kloeckera sp. No. 2201 having high activity of catalase were used as the enzyme materials. The enzyme sources were entrapped in a polyacrylamide gel. Enzymatic properties of the simultaneously immobilized cells were investigated, comparing with those of the mixed whole-cells of the microorganisms. The simultaneously immobilized cells had higher enzyme activity than singly immobilized cells of Pseudomonas polycolor. From this result, the simultaneously immobilized pyridoxine-p oxidase-catalase system could be available to exert a protective effect upon the pyridoxine-p oxidase by destroying $H_2O_2$ which is a by-product of pyridoxine-p oxidation. The optimum pH was 9.0 for the immobilized cells and the whole-cells. The optimum temperature was $45^{\circ}C$ for the immobilized cells and $40^{\circ}C$ for the whole-cells. The pyridoxine-p oxidase of the immobilized cells were activated by $Hg^{++}$ and some SH-compounds.

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The Growth of Proteolytic Bacteria Immobilized in Capsule Type

  • Han Bong-Ho;Choi Su-Il;Kim Seong-Bong;Kim Sang-Ho
    • Fisheries and Aquatic Sciences
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    • v.2 no.1
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    • pp.44-51
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    • 1999
  • Proteolytic bacteria isolated from fermented anchovy jeotkal were immobilized in capsule type with $0.8\%$ sodium alginate and $CaCl_2/carboxymethyl$ cellulose (CMC). For making the immobilized capsule, the optimal concentration of both $CaCl_2$ and CMC, with respect to the membrane hardness and the growth of proteolytic bacteria in capsule, were $2.0\%$ at following conditions: flow rate of $CaCl_2/CMC$ solution and cell suspension were respectively 3.54 ml/min and 0.15 ml/min when inside diameter of inner and outer capillary tube in immobilizing apparatus were 0.32mm, 0.74mm, respectively. The density of proteolytic bacteria in capsule reached maximum, i.e. $10^8-10^9cells$/capsule during culture under optimal conditions in TPY broth, and these were $10^2-10^4$ times higher than these of before culture. During culture of proteolytic bacteria immobilized in capsule type (PBImC) for 72hrs, few growing cells were lost in the outer medium.

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Recovery of Cholesterol from the $\beta$-Cyclodexgtrin-Cholestrerol Complex Using Immobilized Cyclomaltodextrinas of Alkalophilic Bacillus sp. KJ 133

  • Kwon, Ho-Jeong;Jung, Hye-Jin;Kwak, Hae-Soo
    • Journal of Microbiology and Biotechnology
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    • v.11 no.4
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    • pp.712-715
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    • 2001
  • A new combined method including the enzymatic hydrolysis of $\beta$-cyclodextrin ($\beta$-CD) and solvent extraction fo cholesterol from the hydrolyzed mixture was developed to recover cholesterol from a $\beta$-CD-cholesterol complex prepared from dairy products, such as cream, milk, and cheese. Cyclomaltodextrinase (cyclomatodextrin dextrin hydrolase, EC 3.2.1.54, DCase_ prepared form alkalophilic Bacillus sp. KJ 133 hydrolyzed the $\beta$-DC of the $\beta$-CD-cholesterol complex, and then, free cholesterol was efficiently extracted from the hydrolyzed mixture by a nonpolar solvent such as ethyl acetate. To increase the stability of free CDase, immobilized CDase was developed using sodium alginate as a carrier. The immobilized CDase showed a high recovery yield of cholesterol in a time-dependent manner compared to the free CDase. A gas chromatography analysis showed that more than 70% of cholesterol was recovered from the $\beta$-DC-cholesterol complex of cream by the immobilized CDase, whereas only 3% and 29% of cholesterol were recovered when the solvent extraction and free CDase treatment were used, respectively. The cholesterol recovered can be used as a raw material for steroid synthesis. Furthermore, this method can be an efficient way to recover cholesterol or other organic compounds that are bound in a $\beta$ -DC-cholesterol or -organic compound complex.

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The Hydrolysis of Dimethyl-cis-1,3-dibenzyl-2-oxoimidazolidine-4,5-dicarboxylate by Immobilized Whole Cells of Chromobacterium chocolatum (고정화된 Chromobacterium chocolatum의 Whole Cell을 이용한 Dimethyl-cis- 1,3-dibenzyl-2-oxoimidazolidine-4,5-dicarboxylate의 가수분해)

  • Lee, Youn Jin;Shim, Sang Kyun;Ahn, Yong Hyun
    • Journal of the Korean Chemical Society
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    • v.41 no.9
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    • pp.483-487
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    • 1997
  • The whole cells of Chromobacterium chocolatum was immobilized in the matrix of polyacrylamide and then used for the hydrolysis of dimethyl-cis-1,3-dibenzyl-2-oxoimidazolidine-4,5-dicarboxylate. This hydrolysis yielded the optically active monoester ( > 96% ee) which is useful as an synthetic intermediate of (+)-biotin. We have studied the optimum condition of hydrolysis by using immobilized cells under variable concentration of substrate, reaction time and pH levels. The activity of lipase in immobilized cell was retained for longer than 4 weeks. The best conversion yield of product was obtained when 2 g of wet cell was immobilized and then reacted with 200 mg of substrate at pH 7.

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Characterization of Binding of Treponema denticola to Immobilized Fibrinogen using the Fluorescent Fatty Acid Labeling Method

  • Hong, Jin;Lee, Si-Young
    • International Journal of Oral Biology
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    • v.35 no.3
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    • pp.107-111
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    • 2010
  • Treponema denticola is a gram-negative anaerobe that can cause periodontal disease. The adhesion of this bacterium to host tissues is considered to be the primary event in the colonization and infection of a host. Fibrinogen is generally found in damaged tissues resulting from periodontitis. The binding ability of T. denticola to fibrinogen may therefore be an important virulence factor in inducing periodontal diseases. It has been reported recently that oral spirochetes can be labeled with fluorescent fatty acids and we speculated that this labeling method could be used in an oral spirochete binding assay. The binding of several different strains of T. denticola to immobilized human fibrinogen was therefore tested using the fluorescent fatty acid labeling method. In the case of immobilized fibrinogen, the T. denticola ATCC 35405 strain showed saturable binding to immobilized fibrinogen. Indeed, all four different T. denticola strains tested in this experiment, T. denticola ATCC 35405, T. denticola ATCC 33520, T. denticola ATCC 35404 and T. denticola OTK showed binding to fibrinogen. The fluorescent fatty acid labeling method thus shows utility in binding assays for T. denticola, different strains of which can generally bind to immobilized fibrinogen.

The Effect of Temperature on the Stability of Bioluminescence from Immobilized Photobacterium phosphoreum (저장 온도에 따른 고정화 Photobacterium phosphoreum의 Bioluminescence 안정성의 변화)

  • 김현숙;이은수;정성제;유승오;전억한
    • KSBB Journal
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    • v.14 no.1
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    • pp.91-95
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    • 1999
  • The objective of this work was to improve bioluminescence stability of Photobacterium phosphoreum when it stored in view of developing continuous on-line monitoring system for pullutants. Long-term experiments were made to determine the effect of immobilization and storage temperature on the maintenance and stability of bioluminescence from luminescent bacteria. The immobilized cells of P. phosphoreum were compared with free cells in terms of maintenance of bioluminescence at room temperature. The bioluminescence of cells immobilized showed higher bioluminescence intensity that free and strontium bioluminescence stability was investigated with free and immobilized cells stored at $20^{\circ}C,\; 4^{\circ}C,\; -20^{\circ}C\;and\;-70^{\circ}C$for 20 days. Both free and immobilized cells stored at $4^{\circ}C$ emitted a stable bioluminescence while the bioluminescence markedly decreased with those stored at $20^{\circ}C,\;-20^{\circ}C\;and\; -70^{\circ}C$.

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Continuous Production of Cyclodextrin in Two-Stage Immobilized Enzyme Reactor Coupled with Ultrafiltration Recycle System (2단계 고정화 효소반응기를 활용한 Cyclodextrin의 연속생산)

  • Lee, Yong-Hyun;Lee, Sang-Ho;Han, Il-Keun
    • Microbiology and Biotechnology Letters
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    • v.19 no.2
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    • pp.171-178
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    • 1991
  • The two-stage enzyme reactor, packed with cyclodextrin glucanotransferase (CGTase) immobilized on Amberite IRA 900, coupled with ultrafiltration membrane was investigated for continuous production of cyclodextrin (CD). 5% (w/v) of soluble starch was partially cyclized, in the 0.1 l first-stage immobilized enzyme reactor, up to CD conversion yield of 10% (w/w) at retention time of 0.56hr and 1.5 units of immobilized CGTase/1g of carrier. In the second stage main immobilized enzyme reactor capacity of 1.5 l, the maximum CD conversion yield of 39% (w/v) was achieved at retention time of 2.8hr and 0.47 unit of CGTase/1 g of carrier. Unreacted residual dextrin was fractionated with ultrafiltration membrane, and then, recycled into the second-stage main bioreactor to increase the CD conversion yield. The most suitable membrane size and the volume concentration ratio (concentrate: filterate) for recycling of unreacted residual dextrin were found to be 5K dalton and 4:6, respectively. CD conversion yield was increased about 3~4% upon co-immobilization of pulluanase along with CGTase. Spent Amberite IRA 900 can be reutilized consecutively more than 3 times for immobilization of CGTase after regeneration.

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Immobilization of Endo- and Exoinulinase on Vinylsulfone Activated Agarose (Vinylsulfone Activated Agarose 에 Endo- 및 Exoinulinase의 고정화)

  • 한상배;송근섭;정용섭;손희숙;우순자;엄태봉
    • Microbiology and Biotechnology Letters
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    • v.20 no.1
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    • pp.20-24
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    • 1992
  • In order to reuse inulinase effectively, a method for immobilizing both endo- and exoinulinase to vinylsulfone activated agarose via covalent bond was investigated. The immobilized enzyme preparation had, respectively, 400 U for exoinulinase activity and 80 U for endoinu- Iinase activity per gram gel. A thermal stability by immobilization had increased in the case of exoinulinase. Optimum pHs for two immobilized enzymes were 4.4 to 5.0. Synergistic effect which depends on mixed ratio of two immobilized enzymes was the best when the mixed ratio of endo/exo lay between 0.1 and 0.5, and its activity of the mixed enzyme increased 1.7 times as compared to that of each immobilized enzyme. Inulinase activities of both of the immobilized enzymes did not change during 20 times experimental runs in a batch reactor.

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