• Journal of Microbiology and Biotechnology
• /
• v.29 no.5
• /
• pp.749-757
• /
• 2019

Nitrilase is a valuable hydrolase that catalyzes nitriles into carboxylic acid and ammonia. Its applications, however, are severely restricted by the harsh conditions of industrial reaction processes. To solve this problem, a nitrilase from Acidovorax facilis 72W was inserted into an Escherichia coli-Bacillus subtilis shuttle vector for spore surface display. Western blot, enzyme activity measurements and flow cytometric analysis results all indicated a successful spore surface display of the CotB-nit fusion protein. In addition, the optimal catalytic pH value and temperature of the displayed nitrilase were determined to be 7.0 and $50^{\circ}C$, respectively. Moreover, results of reusability tests revealed that 64% of the initial activity of the displayed nitrilase was still retained at the $10^{th}$ cycle. Furthermore, hydrolysis efficiency of upscale production of cyanocarboxylic acid was significantly higher in the displayed nitrilase-treated group than in the free group expressed by E. coli (pET-28a-nit). Generally, the display of A. facilis 72W nitrilase on the spore surface of Bacillus subtilis may be a useful method for immobilization of enzyme and consequent biocatalytic stabilization.

• Korean Chemical Engineering Research
• /
• v.44 no.1
• /
• pp.65-72
• /
• 2006

Recent technical advances in the biorecognition engineering and the microparticle fabrication may enable us to develop the single step purification using magnetic particle, because of its simplicity, efficacy, ease of automation, and process economics. In this study, we used commercial magnetic particles from Seradyn, Inc. (Indianapolis, USA). It was ca. 2.8 micron in diameter, consisted of polystyrene core and magnetite coating, and its surface had carboxyl groups. The model, capture protein was IgG and anti-IgG was used as the ligand molecule. We studied the different surfaces ('nude', ester-activated, and anti-IgG coated) for their biorecognition of IgG. At a high pH condition, we could reduce non-specific binding. Also anti-IgG immobilized magnetic particle could capture IgG more selectively. We attempted 'oriented immobilization' of anti-IgG, in which the polysaccharides moiety near the C-terminus was selectively oxidized and linked to the hydrazine-coated MP, to improve the efficacy of biorecognitive binding. Using this method, the IgG capturing ability was improved by ca. 2 fold. From the binary mixture of the IgG-insulin, IgG could be more selectively captured. In summary, the oriented immobilization of oxidized anti-IgG proved to be as effective as the streptavidin-biotin system and yet simpler and cost-effective. This immobilization method can find its applications in protein biochips and biotargeting.

• KSBB Journal
• /
• v.21 no.5
• /
• pp.384-388
• /
• 2006

The production stability of high-yielding mutants of Streptomyces lincolnensis immobilized on celite beads was examined in repeated batch cultures. We also explored the feasibility of immobilization of vegetative mycelial cells on pre-wetted celite beads, which is practical method for cell immobilization. Repeated transfer of immobilized cells into fresh medium every 10 days increased productivity of immobilized cells and maximum concentration of lincomycin, 1007 $({\pm}256)$ mg/L, was obtained at the end of the ninth cycle. A 1.4-fold higher productivity was obtained in immobilized-cell culture than that obtained by suspended-cell culture. When pre-wetted beads were inoculated with vegetative mycelia and cultured a slightly higher amount of immobilized cells and lincomycin was obtained more than those obtained by culture of spores immobilized on dry beads. This result indicates that immobilization of mycelial cells on pre-wetted beads was readily available. This technique is simple and no additional facilities are required for cell immobilization.

• Korean Chemical Engineering Research
• /
• v.54 no.3
• /
• pp.419-424
• /
• 2016

$H_5PMo_{10}V_2O_{40}$ ($PMo_{10}V_2$) catalyst chemically immobilized on sulfur-containing mesoporous carbon (S-MC) was prepared, and it was applied to the benzyl alcohol oxidation reaction. S-MC was synthesized by a templating method using SBA-15 and p-toluenesulfonic acid as a templating agent and a carbon precursor, respectively. S-MC was then modified to have a positive charge, and thus, to provide sites for the immobilization of $PMo_{10}V_2$. By taking advantage of the overall negative charge of $[PMo_{10}V_2O4_{40}]^{5-}$, $PMo_{10}V_2$ catalyst was immobilized on the S-MC support as a charge matching component. It was revealed that $PMo_{10}V_2$ species were finely and molecularly dispersed on the S-MC via chemical immobilization. In the vapor-phase oxidation of benzyl alcohol, $PMo_{10}V_2$/S-MC catalyst showed higher conversion of benzyl alcohol and higher yield for benzaldehyde and benzoic acid than unsupported $PMo_{10}V_2$ catalyst. The enhanced catalytic performance of $PMo_{10}V_2$/S-MC was due to fine dispersion of $PMo_{10}V_2$ species on the S-MC via chemical immobilization.

• The Journal of Korean Society for Radiation Therapy
• /
• v.17 no.1
• /
• pp.45-55
• /
• 2005

Purpose : To evaluate the usefulness of the handmade patient immobilization device and to report the clinical results of it. Materials and methods : We made two fusion images and analyzed those images. One image is made with diagnostic MR image and CT image, the other with therapeutic planning MR image and CT image. With open head holder, we measured the skin dose and attenuation dose. Also, we made the planning CT couch plate with acrylic plate and styrofoam and compared artifact. Results : We could get more accurate fusion image when we use MR head holder(within 2mm error). The skin dose was reduced 2 times and the attenuation dose was reduced more than $20\%$ when open head holder used. The planning CT couch plate was more convenient than conventional board and reduced artifact remarkably. Conclusion : We could verify the localization point in the MR image which is taken with MR head holder. So we could fuse the image more accurately. The same method could be applied to PET and US image, if the alike immobilization device used. With open head holder, the skin dose and the attenuation dose was reduced. And those above devices could substitute for expensive foreign device, if those are manufactured adequately.

• Korean Journal of Environmental Agriculture
• /
• v.26 no.3
• /
• pp.197-203
• /
• 2007

Zerovalent iron (ZVI) has been widely used in the removal of environmental contaminants from water. The objective of this research was to assess the efficiency of ZVI for immobilization of As (V) in the contaminated water under various chemical conditions. Batch-type experiments showed that the immobilization process followed a first-order kinetic model. Rate constant (k) of the reaction increased consistently and proportionally as increasing ZVI concentrations from 1% (0.158 $hr^{-1}$) to 3% (0.342 $hr^{-1}$), and temperatures from $15^{\circ}C$ (0.117 $hr^{-1}$) to $35^{\circ}C$ (0.246 $hr^{-1}$), respectively. Whereas the rate constant decreased as increasing As (V) concentrations from 1 mg $\Gamma^{-1}$ (0.284 $hr^{-1}$) to 3 mg $\Gamma^{-1}$ (0.153 $hr^{-1}$), and the initial pH from 3 (0.393 $hr^{-1}$) to 9 (0.067 $hr^{-1}$), respectively. Results demonstrated that As (V) in an aqueous solution was rapidly immobilized by ZVI treatments. Zerovalent iron was fast method for remediation of As (V) contaminated water.

• Microbiology and Biotechnology Letters
• /
• v.31 no.4
• /
• pp.368-376
• /
• 2003

Cyclodextrin glucanotransferase (CGTase) from Paenibacillus sp. JB-13 was immobilized on various carriers by several immobilization methods such as ionic binding, covalent linkage and ultrafiltration to improve the process performance. The ultrafiltration and covalent linkage with CNBr-activated sepharose 4B were found as the best method for immobilization of CGTase. The ability of CGTase immobilization onto CNBr-activated sepharose 4B was as high as 18,000 units/g resin when the conditions was as follows: contact time 9 hrs at $37^{\circ}C$, pH 6.0, 100 nm and enzyme loading 24,000 units/g resin. The optimum conditions for production of 2-O-$\alpha$-D-Glucopyranosyl L-Ascorbic acid by immobilized CGTase turned out to be: pH 5.0, temperature $37^{\circ}C$, 20% substrate solution containing 8% (w/v) of soluble starch and 12% (w/v) of L-ascorbic acid sodium salt, 100 rpm, far 25 hrs and with 800 units of immobilized CGTase/ml substrate solution. Moreover the CGTase activity could be stably maintained for 8 times of repetitive reactions after removing products by ultrafiltration through YM 10 membrane.

• KSBB Journal
• /
• v.15 no.1
• /
• pp.49-54
• /
• 2000

Bioluminescence of Photobacterium phosphoreum has been used for the detection of pollutants in the environment. Immobilization method was used to maintain the stability of bioluminescence of P. phosphoreum. The carboxymethylcellulose was investigated to find out whether it was suitable for the immobilization of P. phosphoreum as a matrix without disturbing the bioluminescence emission. A maintenance of bioluminescence was determined from the P. phosphoreum immobilized on the various concentrations of carboxymethylcellulose. A relatively high bioluminescence intensity was shown with immobilized cells on 1%(w/v) carboxymethylcellulose. The effect of carboxymethylcellulose concentrations on the sensitivity of Crcompounds including $Na_{2}CrO_{4}$, $K_{2}CrO_{4}$, $CrO_{3}$, CrK$(SO_4)_{2}$ and $CrCl_{3}$ to the bioluminescence intensity. The calculated $EC_{50}$ showed that the linear relations between such substances and bioluminesence intensity were established.

• 한국생물공학회:학술대회논문집
• /
• 2003.10a
• /
• pp.654-654
• /
• 2003

• Proceedings of the Korean Society for Agricultural Machinery Conference
• /
• 2003.07a
• /
• pp.421-426
• /
• 2003

The Immunosensor based on antigen-antibody binding have been developed for detecting several analytes including antigen, small molecules, and cell. This method can be rapid and show very good detection limits. For Implementation of immunosensor, technologies for immobilization of antibody onto solid surface and detection of protein-protein binding must be developed. (an ellipsis)