• Title/Summary/Keyword: Immobilized cell

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Studies on Whole Cell Immobilized Glucose Isomerase - I. Preparation and Properties of Whole Cell Immobilized Glucose Isomerase - (포도당 이성화 효소의 세포 고정화에 관한 연구 - I. 세포 고정화 효소의 제조와 성질 -)

  • Ahn, Byung-Yoon;Byun, Si-Myung
    • Korean Journal of Food Science and Technology
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    • v.11 no.3
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    • pp.192-199
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    • 1979
  • With cells of Streptomyces spp K-45 isolated from soil, the immobilization of glucose isomerase by a series of treatments ; heat, carefully manipulated drying, extrusion with a thickening agent, and glutaraldehyde-induced crosslinking, was presented. This was aimed to obtain a mechanically stable form of whole cell containing glucose isomerase. The resulted pellet form had a good mechanical strength, compared with a commercial product, and showed 26 % of the activity recovery. The specific activity was 48.1 units per g of the dry material. The immobilized glucose isomerase generally showed properties similar to those of the soluble enzyme ; optimal pH at $7.5{\sim}9.0$, optimal temperature at $80{\sim}85^{\circ}C$, activation energy of 10.9 kcal/mole, and $K_m$ for glucose of 10.9M. The immobilized enzyme was very thermostable and pH stable.

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Studies on Microbial Penicillin Amidase (II) Characteristics and the Reactor Performance of Whole Cell Immobilized Penicillin Amidase of Escherichia coli (미생물 페니실린 아미다제에 관한 연구 (II) E. coli의 균체 고정화 페니실린 아미다제의 특성 및 반응조에 관한 연구)

  • Seong, Baik-Lin;Kim, Bong-Hee;Mheen, Tae-Iek;Moon H. Han
    • Microbiology and Biotechnology Letters
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    • v.9 no.1
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    • pp.35-44
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    • 1981
  • Whole cell penicillin amidase of Escherichia coli was immobilized by entrapment in gelatin followed by extrusion and crosslinking with glutaraldehyde. The immobilized engyme preparation demonstrated the recovery yield of activity up to 70% and good stability during storage and operation. The half life of activity decay during the operation was estimated to be about 50 days. The optimum pH and temperature for both of immobilized and soluble enzyme are 8.5 and 5$0^{\circ}C$, respectively. No significant change was demonstrated in the effect of pH and temperature, but the increase in heat stability at high temperature was observed in the case of the immobilized enzyme. It was found that the plug flow reactor could be operated favorably since the pH drop along the column path due to tile reaction product was minimized by employing substrate solution with moderate buffer strength. The optimal condition of reactor operation was discussed with regard to the effect of substrate concentration and the residence time on the conversion efficiency and productivity.

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Production of casein phosphopeptides using Streptococcus faecalis var. liquefaciens cell immobilization (Streptococcus faecalis var. liquefaciens 전세포 고정화법을 이용한 Casein Phosphopeptides 생산)

  • Lee, Ki-Sung;Shin, Jae-Yoon;Jang, Yi-Hyun;Kweon, Dae-Hyuk;Park, Ki-Moon;Jin, Yong-Su
    • KSBB Journal
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    • v.23 no.1
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    • pp.59-64
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    • 2008
  • Optimum conditions for production of casein phosphopeptides (CPP) from sodium casenate by immobilized cell culture of Streptococcus faecalis var. liquefaciens were investigated. Immobilized cells were made by mixing 60% sodium alginate solution with an equal volume of culture broth at the end of exponential phase and subsequently dropping the mixture into $CaCl_{2}$ solution. Optimum conditions for CPP production by the immobilized cells were the same as those ($50^{\circ}C$, pH 7.0, and 10% substrate concentration) by the crude enzyme solution from the supernatant of culture broth. Optimum loading volume of the immobilized cells into a batch reactor was 30% (w/v). Using a continuous reactor loaded by the immobilized cells under the identified optimal conditions, we were able to produce CPP continuously up to 30 days with a maximum CPP conversion efficiency of 20%.

Activation and immobilization of phenol-degrading bacteria on oil palm residues for enhancing phenols degradation in treated palm oil mill effluent

  • Tosu, Panida;Luepromchai, Ekawan;Suttinun, Oramas
    • Environmental Engineering Research
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    • v.20 no.2
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    • pp.141-148
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    • 2015
  • The presence of phenols in treated palm oil mill effluent (POME) is an environmental concern due to their phytotoxicity and antimicrobial activity. In this study, phenol-degrading bacteria, Methylobacterium sp. NP3 and Acinetobacter sp. PK1 were immobilized on oil palm empty fruit bunches (EFBs) for removal of phenols in the treated POME. The bacterial exopolysaccharides (EPS) were responsible for cell adhesion to the EFBs during the immobilization process. These immobilized bacteria could effectively remove up to 5,000 mg/L phenol in a carbon free mineral medium (CFMM) with a greater degradation efficiency and rate than that with suspended bacteria. To increase the efficiency of the immobilized bacteria, three approaches, namely activation, acclimation, and combined activation and acclimation were applied. The most convenient and efficient strategy was found when the immobilized bacteria were activated in a CFMM containing phenol for 24 h before biotreatment of the treated POME. These activated immobilized bacteria were able to remove about 63.4% of 33 mg/L phenols in the treated POME, while non-activated and/or acclimated immobilized bacteria could degrade only 35.0%. The activated immobilized bacteria could be effectively reused for at least ten application cycles and stored for 4 weeks at $4^{\circ}C$ with the similar activities. In addition, the utilization of the abundant EFBs gives value-added to the palm oil mill wastes and is environmentally friendly thus making it is attractive for practical application.

Biodetoxification of Coumaphos Insecticide Using Immobilized Escherichia coli Expressing Organophosphorus Hydrolase Enzyme on Cell Surface

  • Mansee, Ayman H.;Chen, Wilfred;Mulchandani, Ashok
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.5 no.6
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    • pp.436-440
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    • 2000
  • Recently, we reported an improved technology for the degradation of organophosphate nerve agents using whole cells of genetically engineered Escherichia coli that anchored and displayed the enzyme organophosphorus hydrolase on the cell surface. In this paper we report the immobilization of these cells on highly porous sintered glass beads and the subsequent application of the immobilized cell in a continuous-flow packed bed bioreactor for the biodetoxification of a widely used insecticide, coumaphos.

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Detection of ${\alpha}-Cyclodextrin$ and E.coli Cell Using Polydiacetylene Supramolecules

  • Lee, Gil-Sun;Choi, Hyun;Lee, Chung-Wan;Ahn, Dong-June;Oh, Min-Kyu;Kim, Jong-Man
    • Proceedings of the Polymer Society of Korea Conference
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    • 2006.10a
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    • pp.306-306
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    • 2006
  • We immobilized and patterned PDA vesicles on solid substrate using micro arrayer, which have moieties to react with chemical and biological materials. Immobilized vesicle system was developed since it possesses many advantages in multiple screening, durable stability, and higher sensitivity. We applied polydiacetylene supramolecules to chemical and biological sensors for detection of ${\alpha}-cyclodextrin$ and E.coli cell selectively. This detection method could be applied as DNA chip, protein chip, and cell chip for multiple screening as well as chemical sensor by modifying the functional groups of diacetylene monomer.

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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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Production of (R)-Ethyl-4-Chloro-3-Hydroxybutanoate Using Saccharomyces cerevisiae YOL151W Reductase Immobilized onto Magnetic Microparticles

  • Choo, Jin Woo;Kim, Hyung Kwoun
    • Journal of Microbiology and Biotechnology
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    • v.25 no.11
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    • pp.1810-1818
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    • 2015
  • For the synthesis of various pharmaceuticals, chiral alcohols are useful intermediates. Among them, (R)-ethyl-4-chloro-3-hydroxybutanoate ((R)-ECHB) is an important building block for the synthesis of L-carnitine. (R)-ECHB is produced from ethyl-4-chloro-3-oxobutanoate (ECOB) by a reductase-mediated, enantioselective reduction reaction. The Saccharomyces cerevisiae YOL151W reductase that is expressed in Escherichia coli cells exhibited an enantioselective reduction reaction toward ECOB. By virtue of the C-terminal His-tag, the YOL151W reductase was purified from the cell-free extract using Ni2+-NTA column chromatography and immobilized onto Ni2+-magnetic microparticles. The physical properties of the immobilized reductase (Imm-Red) were measured using electron microscopy, a magnetic property measurement system, and a zeta potential system; the average size of the particles was approximately 1 μm and the saturated magnetic value was 31.76 emu/g. A neodymium magnet was used to recover the immobilized enzyme within 2 min. The Imm-Red showed an optimum temperature at 45℃ and an optimum pH at 6.0. In addition, Bacillus megaterium glucose dehydrogenase (GDH) was produced in the E. coli cells and was used in the coupling reaction to regenerate the NADPH cofactor. The reduction/oxidation coupling reaction composed of the Imm-Red and GDH converted 20 mM ECOB exclusively into (R)-ECHB with an e.e.p value of 98%.

Immobilization of Glucose Oxidase on Multi-Wall Carbon Nanotubes for Biofuel Cell Applications

  • JUNG SOO KEUN;CHAE YOUNG RAE;YOON JONG MOON;CHO BYUNG WON;RYU KEUN GARP
    • Journal of Microbiology and Biotechnology
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    • v.15 no.2
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    • pp.234-238
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    • 2005
  • Glucose oxidase was immobilized on the carboxylated multi-wall carbon nanotubes (MWNT-COOHs) in the presence of a coulping reagent, 1-ethy1-3-(3-dimethylaminopropy1) carbodiimide. Significant amounts of glucose oxidase were also immobilized on MWNT-COOHs without the coupling reagent. Various conditions for the immobilization of glucose oxidase were optimized. Optimal pH for the maximal activity of the immobilized glucose oxidase shifted to 7 from the optimal pH of 6 for the maximal activity of free enzyme due to the carboxy1 groups on the surface of MWNT-COOHs. An electrode of graphite rod with a diameter of 6 mm was fabricated using the immobilized glucose oxidase. The cyclic voltammetry study of the enzyme electrode revealed that the oxidation of glucose and subsequent transfer of electrons from the oxidation of glucose to the electrode were possible by the immobilized glucose oxidase without a mediator, implying that the enzyme electrode can be utilized for the development of biofuel cells.

Hydrogen Production by the Immobilized Cells of Rhodopseudomonas sp. E15-1 (Rhodopseudomonas sp. E15-1의 균체 고정화에 의한 수소생성)

  • Bae, Moo;Park, Sun-Hee
    • Microbiology and Biotechnology Letters
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    • v.17 no.1
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    • pp.74-80
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    • 1989
  • For improvement of photobiological hydrogen production, Rhodopseudomonas El5-1, a photo-synthetic becterium capable of producing n high yield of hydrogen, was immobilized and conditions for hydrogen production by immobilized cells were examined. The optimum concentration for the combined matrix was obtained when sodium alginate was used at final concentration of 4%. The immobilized cells may reduce the inhibitory effects of nitrogen or oxygen. To minimize the diffusion resistance of the nutrients in alginate gel, the bend size less than 2 mm in diameter was desirable. The immobilized cells were also able to utilize n wide range of organic substrates for the production of hydrogen. The hydrogen producing activity of the immobilized cells was maintained for 20 days without loss of activity during semi-continuous operation of the reactor by feeding of new medium periodically and continuous production of hydrogen could be successfully performed for 30 days.

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