• Journal of Microbiology and Biotechnology
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• v.14 no.3
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• pp.584-592
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• 2004

A thermostable $\beta$-glycosidase gene, tfi $\beta$-gly, was cloned from the genomic library of Thermus filiformis Wai33 A1. ifi $\beta$-gly consists of 1,296 bp nucleotide sequence and encodes a polypeptide of 431 amino acids. It shares a strong amino acid sequence similarity with the $\beta$-glycosidases from other Thermus spp. belonging to the glycosyl hydrolase family 1. In the present study, the enzyme was overexpressed in Escherichia coli BL21 (DE3) using the pET21b(+) vector system. The recombinant enzyme was purified to homogeneity by heat treatment and a $Ni^{2+}$-affinity chromatography. Polyacrylamide gel electrophoresis (PAGE) showed that the recombinant Tfi $\beta$-glycosidase was a monomeric form with molecular mass of 49 kDa. The temperature and pH range for optimal activity of the purified enzyme were 80- $90^{\circ}C$ and 5.0-6.0, respectively. Ninety-three percent of the enzyme activity was remained at $70^{\circ}C$ after 12 h, and its half-life at $80^{\circ}C$ was 6 h, indicating that Tfi $\beta$-glycosidase is highly thermostable. Based on its K_m$, or $K_{cat}K_m$, ratio, Tfi $\beta$-glycosidase appeared to have higher affinity for $\beta$-D-glucoside than for $\beta$-D-galactoside, however, $K_{cat} for \beta$-D-galactoside was much higher than that for $\beta$-D-glucoside. The activity for lactose hydrolysis was proportionally increased at $70^{\circ}C$ and pH 7.0 without substrate inhibition until reaching 250 mM lactose concentration. The specific activity of Tfi TEX>$\beta$-glycosidase on 138 mM lactose at $70{^\circ}C$ and pH 7.0 was 134.9 U/mg. Consequently, this newly cloned enzyme appears to have a valuable advantage of conducting biotechnological processes at elevated temperature during milk pasteurization in the production of low-lactose milk.

• Nuclear Engineering and Technology
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• v.27 no.6
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• pp.877-887
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• 1995

YGN 3 is the first nuclear power plant to use the Core Protection Calculator (CPC) as the core protection system and the Core Operating Limit Supervisory System (COLSS) as the core monitor-ing system in Korea. The CPC is designed to provide on-line calculations of Departure from Nucleate Boiling Ratio (DNBR) and Local Power Density (LPD) and to initiate reactor trip if the core conditions exceed the DNBR or LPD design limit. The COLSS is designed to assist the operator in implementing the Limiting Conditions for Operation (LCOs) in Technical Specifications for DNBR/Linear Heat Rate (LHR) margin, azimuthal tilt, and axial shape index and to provide alarm when the LCOs are reached. During YGN 3 initial startup testing, extensive CPC/COLSS related tests ore peformed to ver-ify the CPC/COLSS performance and to obtain optimum CPC/COLSS calibration constants at var, -ious core conditions. Most of test results met their specific acceptance criteria. In the case of missing the acceptance criteria, the test results ore analyzed, evaluated, and justified. Through the analysis and evaluation of each of the CPC/COLSS related test results, it can be concluded that the CPC/COLSS are successfully Implemented as designed at YGN 3.

• The Journal of the Korean Society for Microbiology
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• v.21 no.1
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• pp.151-161
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• 1986

In order to develop sensitive and sepcific assay methods for E. coli heat labile enterotoxin(LT) hybridoma cell lines secreting LT specific monoclonal antibody were obtained. LT was purified from cell lysate of E. coli O15H11. The steps included disruption of bacteria by French pressure, DEAE Sephacel ion exchange chromatography, Sephadex G200 gel filtration, and second DEAE Sephacel ion exchange chromatography, successively. Spleen cells from Balb/c mice immunized with the purified LT and $HGPRT^{(-)}$ plasmacytomas, $P3{\times}63Ag8.V653$ were mixed and fused by 50% (w/v) PEG. Hybrid cells were grown in 308 wells out of 360 wells, and 13 wells out of them secreted antibodies reacting to LT. Among these hybridoma cell 1G8-1D1 cell line was selected since it had produced high-titered monoclonal antibody continuously. By using culture supernatant and ascites from 1G8-1D1 cells the monoclonal antibody was characterized, and an assay system for detecting enterotoxigenic E. coli was established by double sandwich enzyme-linked immunosorbent assay (ELISA). The following results were obtained. 1. Antibody titers of culture supernatant and ascites from 1G8-1D1 hybridoma cells were 512, and 102, 400, respectively by GM1-ELISA and its immunoglobulin class was IgM. 2. The maximum absorption ratio of 1G8-1D1 cell culture supernatant to LT was 90% at $300\;{\mu}g/ml$ of LT concentration. LT concentration shown at 50% absorption ratio was $103.45{\mu}g$ and the absorption ratio was decreased with tile reduction of LT concentration. This result suggests that monoclonal antibody from 1G8-1D1 hybridoma cell bound with LT specifically. 3. The reactivities of 1G8-1D1 cell culture supernatant to LT and V. cholerae enterotoxin(CT) were 0.886 and 0.142(O.D. at 492nm) measured by the GM1-ELISA, indicating 1G8-1D1 monoclonal antibody reacted specifically with LT but not with CT. 4. The addition of 0.1ml of ascites to 0.6mg and 0.12mg of LT decreased the vascular permeability factor to 41% and 44% respectively, but it did not completely neutralize LT. 5. By double sandwich ELISA using monoclonal antibody, as little as 75ng of the purified LT per ml could be detected. 6. The results by assay of detecting LT in culture supernatants of 14 wild strains E. coli isolated from diarrhea patients by the double sandwich ELISA were almost the same level as those by reverse passive latex agglutination.

• Journal of the Korean Recycled Construction Resources Institute
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• v.1 no.1
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• pp.89-97
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• 2005

Recently, for the problem solution of demand and supply imbalance of fine aggregate due to the shortage of natural fine aggregate resource and the environment regulation on sea sand extraction in the construction field, the studies for the application of recycled fine aggregate using waste concrete are being progressed versatilely. On the other hand, the treatment of fly-ashes that of industrial by-product originated in the steam power plant is discussed by the continuous increasing of origination quantities. In the ease of using fly-ash, advantages are the improvement of workability, viscosity and long-time strength, and the reduction of hydration heat under the early ages, as the admixtures for concrete, but the studies for the application of fly-ash as recycled concrete admixtures are inadequacy. There fore, in this study, through investigating the properties of fresh, hardened and durability according to the replacement of recycled fine aggregate and fly-ash, it is intended to propose the fundamental data for structural application of recycled concrete using recycled fine aggregate and fly-ash. As the result of this study, they arc shown that the engineering properties and durability, in the case of replacement ratio 100% of recycled fine aggregate, arc similar to those of concrete using natural fine aggregate, so it is considered that recycled fine aggregate could be used as the fine aggregate for concrete. Also, the performances of recycled concrete are improved by replacing fly-ash.