• Proceedings of the Korean Society for Applied Microbiology Conference
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• 1986.12a
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• pp.509.1-509
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• 1986

Recent advances in recombinant DNA techniques have provided a tool for breeding of microorganisms of hyper production. Enzyme production by cloned microorganism has some advantages. They are ⅰ) Enzymes can be produced by a microorganism easily cultured ⅱ) Hyper production. ⅲ) In some cases, such as thermophilic enzyme gene is cloned in a mesophilic bacteria, the enzyme purification procedure can be simplified. One example, production of thermophilic ${\beta}$-galactosidase in B. subtilis will be presented. Bacillus stearothermophilus IAM 11001 produced three ${\beta}$-galactosidases, ${\beta}$-galactosidase I, II and III (${\beta}$-gal-I, II and III). By connecting restriction fragments of the chromosomal DNA to plasmid vector, followed by transformation of Escherichia coli, two ${\beta}$-galactosidase genes (bgaA and bgaB) located close to each other on the chromosome were cloned.

• KSBB Journal
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• v.27 no.6
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• pp.341-346
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• 2012

Indican (indoxyl-${\beta}$-D-glucoside) is a colorless natural compound and can be used as a precursor for the production of indigo. This production step only require an enzyme, ${\beta}$-glucosidase, that readily screened from microbial resource by using selective media supplemented with indican as a sole carbon source. Agrobacterium tumefaciens was well grown in this media and thus presumed to produce a related enzyme. The corresponding gene, encoding a protein with a calculated molecular mass of 51 kDa, was cloned and overexpressed as MBP fusion proteins. The purified enzyme was determined to be a dimer and showed the maximum activity for indican at pH 7.0 and $40^{\circ}C$. The kinetic parameters for indican, Km and Vmax, were determined to be 1.4 mM and 373.8 ${\mu}M/min/mg$, respectively. The conversion yield of indican into indigo using this enzyme was about 1.7-1.8 folds higher than that of previously isolated enzyme from Sinorhizobium meliloti. Additionally, this enzyme was able to hydrolyze various ${\beta}$-1,4 glycoside substrates.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1997.04a
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• pp.93-93
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• 1997

Analysis of protein is often frustrated by the inability to isolate large amounts of purified protein from a native source. To overcome this problem, fusion protein expression systems such as pGEX system have been widely used. Using pGEX system, the desired protein could be easily obtained in a large amount in E. coli, and then the fusion protein could be used for the study of the function of the given protein. To analyze and purify the GST fusion protein, anti-GST antibody could be used as one of the system of choice. However, the production and characterization of monoclonal anti-GST antibody has not been studied extensively yet. To produce monoclonal anti-GST antibody, GST was purified from E. coli transformed with pGEX-cs, one of the pGEX system and was used as an antigen. The monoclonal antibody was produced by fusion of the immunized spleen cells with SP2-0 myeloma cells. The antibody was characterized by ELISA, western blotting, etc. The monoclonal antibody produced in this study (mAb-GSTA) showed strong and specific immunoreactivity against not only GST but also GST-fusion proteins. Also, mAb-GSTA was successfully used for the immunoaffinity purification of the GST ${\beta}$-Rc.-third intracellular-loop fusion protein. The results of the present study suggest that mAb-GSTA may be used for the identification and purification of GST fusion proteins.

• Asian-Australasian Journal of Animal Sciences
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• v.23 no.10
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• pp.1369-1379
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• 2010

This study investigated purification and milk-clotting activity of the enzymes produced by Bacillus subtilis var, natto and Rhizopus oligosporus compared with that of commercial rennet. The clotting time, viscosity, tension and microstructure of the curd and electrophoretic patterns of milk proteins were determined. The milk-clotting activity/proteolytic activity ratios (MCA/PA ratio) of B. subtilis, R. oligosporus and commercial rennet were also compared. The results revealed that the curd formed by the commercial rennet had the highest viscosity and curd tension and the shortest clotting time among the three enzymes. However, curd produced by Rhizopus enzymes was ranked as second. From the MCA/PA ratio and electrophoretogram analyses it could be concluded that the enzyme produced by B. subtilis had the highest proteolytic activity, while the commercial rennet had the highest milk-clotting activity. Observations of microstructures of SEM showed that the three-dimensional network for curd formed by commercial rennet was denser, firmer and more smooth. The milk-clotting activity, specific activity, purification ratio and recovery of the purified enzymes produced by both the tested organisms were also determined with ion exchange chromatography and gel filtration.

• The Plant Pathology Journal
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• v.20 no.2
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• pp.135-141
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• 2004

A filamentous virus was isolated from taro (Colocasia esculenta Schott) showing mosaic and chlorotic feather-ing symptoms in Chuncheon, Gangwon province in 2002. Based on ELISA, its appearance in electron microscope, serological relationships, and RT-PCR using specific primer and nucleotide sequence analysis of the CP gene, the isolated virus was identified as Dasheen mosaic virus (DsMV) and designated as Korean isolated (DsMV-Kr). DsMV was not serologically related to Zantedeschia mosaic virus (ZaMV), which has been reported to infect an Araceae plants. Since the coat protein revealed electrophoretic heterogeneity, about 42 kDa, 39 kDa and 31 kDa by SDS-PAGE, an improved purification method was established for the production of antisera against DsMV-Kr. The purification method used in this study may be effectively applied to the purification of other filamentous viruses.

• Microbiology and Biotechnology Letters
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• v.20 no.2
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• pp.178-182
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• 1992

For an industrial-scale purification and production of cephalosporin C from a culture broth of Ceplzalos#mium nmemonium CSA-2.8A3 mutant, ultrafiltration, column chromatography, reverse osmosis, and spray drying were empolyed. Above 90% of yield and high purity of cephalosporin C were obtained through WA-30, HP-20, XAD-2000 and SK-1B column chromatographies. Especially, in the tendom operation of the columns, the recovery yield was increased up to 96%. The purified cephalosporin C was stable at $4^{\circ}C$ and in acidic condition, while it was unstable at room temperature and in alkaline condition at pH above 8.0. Cephalosporin C powder or a final product prepared by spray drying contained 85.554 of sodium cephalosporin C, 6.3%' of water, 4.63% of free $Na^+$ ions. and traces of metal ions.

• Proceedings of the Korean Institute of Information and Commucation Sciences Conference
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• 2022.10a
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• pp.43-46
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• 2022

The optimal operation of the water purification plant can be carried out only when the required flow rate is supplied in a timely manner using the minimum electrical energy by accurately predicting the pattern and amount of tap water used in the consumer. In order to ensure the stability of tap water production and supply, a system that can be pre-verified before applying AI-based composite sensors to the water purification plant was established to derive complementary matters through the pre-verification model for each composite sensor and improve the quality and operation stability of the composite sensor data.

• Journal of the Korean Magnetic Resonance Society
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• v.15 no.1
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• pp.25-39
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• 2011

Syndecan-4 is a transmembrane heparan sulfate proteoglycan, which is a coreceptor with integrins in cell adhesion. To get better understand the mechanism and function of Syndecan-4, it is critical to elucidate the three-dimensional structure of a single transmembrane spanning region of them. Unfortunately, it is hard to prepare the peptide because syndecan-4 is membrane-bound protein that transverse the lipid bilayer of the cell membrane. Generally, the preparation of transmembrane peptide sample is seriously difficult and time-consuming. In fact, high yield production of transmembrane peptides has been limited by experimental adversities of insufficient yields and low solubility of peptide. Here, we demonstrate experimental processes and results to optimize expression, purification, and NMR measurement condition of Syndecan-4 transmembrane peptide.

• Progress in Medical Physics
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• v.27 no.3
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• pp.156-161
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• 2016

When air discharged from a radioisotope production facility is contaminated with radiation, the public may be exposed to radiation. The objective of this study is to manage such radiation exposure. We measured the airborne radioactivity concentration at a 30 MeV cyclotron radioisotope production facility to assess whether the exhaust gas was contaminated. Additionally, we investigted the radioactive contamination of the air filter for efficient air purification and radiation safety control. To measure the airborne radiation concentration, specimens were collected weekly for 4 h after the beginning of the radioisotope production. Regarding the air purifier, five specimens were collected at different positions of each filter-pre-filter, high-efficiency particulate air filter, and charcoal filter-installed in the cyclotron production room. The concentrations of F-18, I-123, I-131, and Tl-201 generated in the radioiodine production room were $13.5Bq/m^3$, $27.0Bq/m^3$, $0.10Bq/m^3$, and $11.5Bq/m^3$, respectively; the concentrations of F-18, I-123, and I-131 produced in the radioisotope production room were $0.05Bq/m^3$, $16.1Bq/m^3$, and $0.45Bq/m^3$, correspondingly; and those of F-18, I-123, I-131, and Tl-201 generated in the accelerator room were $2.07Bq/m^3$, $53.0Bq/m^3$, $0.37Bq/m^3$, and $0.15Bq/m^3$, respectively. The maximum radiation concentration of I-123 generated in the radioiodine production room was 1,820 Bq/g, which can be disposed after 2 days. The maximum radiation concentration of Tl-202 generated in the radioisotope production room was 205 Bq/g, and this isotope must be stored for 53 days. The I-123 generated in the radioiodine production room had a maximum concentration of 1,530 Bq/g and must be stored for 2 days. The maximum radiation concentration of Na-22 generated in the radioisotope production room was 0.18 Bq/g and this isotope must be disposed after 827 days. To manage the exhaust, the efficiency of air purification must be enhanced by selecting an air purifier with a long life and determining the appropriate replacement time by examining the differential pressure through systematic measurements of the airborne radiation contamination level.

• Journal of the Korean Society of Food Science and Nutrition
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• v.27 no.2
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• pp.333-337
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• 1998

This study was carried out to get infomation on the nitric oxide production ability and its formation mechanisms of polysaccharides extracted from Ganoderma lucidum(PSG) by using murine macrophage cell line. The cultured mycelial cells of Ganoderma lucidum were extracted by alkali, and than neutralized by acid. The extract were passed through the column of DEAE cellulose for more purification. The neutral fraction was concentrated and precipitated with 95% ethanol. The precipitate was lyophilized and PSG was obtained. The immunomodulating effects of PSG on macrophage were performed by using murine macrophage cell line ATCC TIB 71 cells with PSG 0.5mg. PSG alone could not induce the production of nitrite, but it had a significant potential effect on nitrite secretion when the cells were primed and triggered with BCG and Interferon(IFN)-${\gamma}$. Also it was prominent by using calcium channel blocker(verapamil) and adenylate cyclase activator(forskolin).