• Microbiology and Biotechnology Letters
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• v.20 no.5
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• pp.519-526
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• 1992

The gene coding for urease of alkalophilic Bacillus pasteurii had been cloned in Escherichia coli previously. The urease protein was purified 63.1-fold by TEAE-cellulose, DEAE-Sephadex A-50, Sephadex G-150 and Sephadex G-200 chromatographies with a 7.3% yield from the sonicated fluid of the E. coli HB1Ol(pBUll) encoding B. pasteurii urease gene. The ureases of E. coli (pBUll) and B. pasteurii possessed as a $K_m$ for urea, 42.1 mM and 40.4 mM, respectively. They hydrolyzed urea with $V_{max}$ of 86.9$\mu$mol/min and 160$\mu$mol/min, respectively. Both ureases were composed with four subunits (Mrs 67,000) and a subunit (Mr 20,000). The molecular weight of both native enzymes was Mr 280,OOO$pm$10,000 determined by gel filtration chromatography and Coomassie blue staining of the subunits. The optimal reaction pH of both ureases were pH 7.5. The ureases were stabled in pH 5.5-10.5. The optimal reaction temperature of both ureases were $60^{\circ}C$, and the ureases were stable for an hour at $50^{\circ}C$, 40min at $60^{\circ}C$ and 10 min at $70^{\circ}C$ The activity of both enzymes were inhibited completely by $Ag^{2+}$, $Hg^{2+}$, $Zn^{2+}$, $Cu^{2+}$, and were inhibited 60% by CoH, 30% by $Fe^{2+}$ and 10% by $Pb^{2+}$. However it was increased by the addition of $Sn^{2+}$, $Mn^{2+}$, $Mg^{2+}$ at concentration of $1{\times}10^{-3}$M. Both ureases were inhibited completely by p-CMB and acetohydroxamic acid. The urease expressed in E. coli (pBU11) was inhibited 70% by SDS. The urease of B. pasteurii was inhibited 40% by hydroxyurea, whereas the recombinant urease of E. coli strain was inhibited 17%. Both enzymes were not inhibited by cyclohexanediaminetetraacetic acid (CDTA) and ethylendiaminetetraacetic acid (EDTA).

• Korean Journal of Microbiology
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• v.48 no.2
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• pp.109-115
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• 2012

Helicobacter pylori is an important factor of chronic gastritis, digestive ulcer, and stomach cancer. CagL, a virulence factor of H. pylori, is well-known as a pilus protein which acts as adhesion to host cell and a component of Type 4 secretion system. In this study, we evaluated the protective response of recombinant CagL protein (rCagL) using Mongolian gerbil animal model for H. pylori infection. The cagL gene was cloned from 26695 H. pylori followed by over-expression and purification of the protein in E. coli. Mongolian gerbils were immunized with rCagL protein mixed with aluminum adjuvant via intramuscular injections once a week during 4 weeks. At a week after the last immunization, the Mongolian gerbils were administrated with H. pylori 7.13 strain into the stomach and sacrificed to measure antibody titer on rCagL by ELISA and bacterial colonization in the stomach, and to examine the histopathological changes and cytokine expression at 6 week after challenge. Antibody titers on recombinant protein were significantly increased from a week after the first immunization. There was no significant change of the number of bacterial colony between control group and immunized group. The relative stomach weight was significantly decreased in immunized group, but the significant change of histopathological assessment was not observed in the stomach. Cytokine expression such as IL-$1{\beta}$ and KC also was not significantly different between control and immunized groups. These results indicate that rCagL could effectively induce the formation of the specific IgG antibodies. However, bacterial colonization and histopathological lesions could not be inhibited by the immunization in the stomach, indicating not enough protection against H. pylori infection. We consider that along with CagL other adequate antigens could be needed stimulating immune response and inducing protective effects against gastric disease, and also a better adjuvant could be considered.

• Journal of the Korea Concrete Institute
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• v.21 no.1
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• pp.85-92
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• 2009

A concrete is considered unfriendly-environmental material because it uses cement which emits much $CO_2$ during producing process. However, a concrete absorbs $CO_2$ through carbonation process during service life. In this paper how much concrete absorbs $CO_2$ through carbonation was calculated using 1) concentration of carbonatable substances in concrete, 2) carbonated volume of concrete, 3) molecular weight of $CO_2$ based on references and the method was proposed. $CO_2$ emission from producing $1m^3$ concrete was calculated based on $CO_2$ emission datum of materials used in concrete. From using these methods that calculate $CO_2$ emission and absorption of concrete, assessment of $CO_2$ emission-absorption against a real apartment was conducted by subtracting absorption $CO_2$ according to service life from $CO_2$ emission in the process of making concrete. As a result, a ratio of absorption over emission of $CO_2$ through concrete carbonation according to service life 40, 60, 80 years was assessed about 3.65, 4.47, 5.18%. An objective of this study is to propose how to calculate emission - absorption of $CO_2$ from producing and using concrete. Although the result value, emission - absorption of $CO_2$, is 5.18% very low when the service life of an apartment is 80years, the value can be improved by reducing emission from using blended cement such as blast furnace slag or increasing replacement ratio of cement and increasing carbonated volume of concrete from expanding service life of a building. This study may be useful when $CO_2$ emission - absorption of concrete is evaluated in the further study.

• Journal of the Korean Society of Food Science and Nutrition
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• v.33 no.1
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• pp.52-58
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• 2004

The active principle responsible for lipid lowering in Chinese cabbage kimchi, 3-(4-hydroxyl-3'5'-dimethoxyphenyl)propionic acid, of molecular weight 226, was chemically synthesized and then used for the treatment of hypercholesterolemia in male New Zealand white rabbit. Hypercholesterolemia in rabbits were induced by feeding 0.5% cholesterol added chow diet for 5 weeks. Each experimental group has four rabbits in it. for the 1st experiment,3-(4-hydroxyl-3'5'-dimethoxyphenyl)propionic acid or simvastatin was injected to the ear vein of rabbit every other day for 16 days (2 mg/3 kg/2 days) while normal chow diet was provided. Blood was drawn every 4th day. For the 2nd experiment, all the experimental condition was same as the 1st trial except 0.5% cholesterol diet was provided while 16 days. Plasma cholesterol level was decreased when cholesterol in the diet was removed. Decreased in cholesterol in kimchi and simvastatin groups were 18.65 and 29.67%, respectively compared to the control when the normal diet was given, and cholesterol increase was inhibited by 33.79 and 21.81% for kimchi and simvastatin groups, respectively, when 0.5% cholesterol diet was provided. The drop in LDL-C level by the active principle of kimchi and simvastatin was not significant when normal diet was given, however the changes was significant (p<0.05), approximately 130% decrease, when 0.5% cholesterol diet was given. The 105% and 62% decreased in triglyceride concentration were observed from 0.5% cholesterol diet fed kimchi and simvastatin groups respectively HDL cholesterol levels in experimental groups were not changed significantly from the both trials. The HMG-CoA reductase activity of kimchi and simvastatin groups were found to be higher than that of control to compensate the hypercholesterolemic condition induced by 0.5% cholesterol diet in these groups. In conclusion, diet is an important factor to control the hypercholesterolemia besides drug treatment. 3-(4-hydroxyl-3'5'-dimethoxyphenyl)propionic acid that is the active principle in Chinese cabbage kimchi seems a beneficial to the hypercholesterolemia and its effect is comparable to that of simvastatin.

• Journal of the Korean Society of Food Science and Nutrition
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• v.41 no.12
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• pp.1764-1770
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• 2012

This study was performed to investigate the changes of total and soluble ${\beta}$-glucan contents, purity, and physical characteristics of three heated barley varieties: Saessalbori (SSB), Saechalssalbori (SCSB), and Hinchalssalbori (HCSB). The barleys were heated at different temperatures of 110, 120, 130, 140 and $150^{\circ}C$ for 2 hours. The total ${\beta}$-glucan contents of raw SSB, SCSB, and HCSB were 8.40, 7.77 and 8.28%, and the soluble ${\beta}$-glucan contents were 4.79, 4.14, and 4.61%, respectively. After heating at $130^{\circ}C$, the total ${\beta}$-glucan contents increased to 11.59, 14.6, and 13.36%, as did the soluble ${\beta}$-glucan contents to 4.21, 7.96, and 7.23%, respectively. The purities of soluble ${\beta}$-glucan of the raw barleys were 35.11, 32.74 and 25.62%, but after heating at $150^{\circ}C$, it increased to 83.43, 91.02, and 88.01%, respectively. The molecular weight and viscosity of the ${\beta}$-glucan solution decreased with increasing heating temperature. The re-solubility of raw barley ${\beta}$-glucan was about 50%, but it was increased to 97% with increasing heating temperature. These results suggest that heating of ${\beta}$-glucan can improve the utilization of barley ${\beta}$-glucan.

• Journal of Dental Rehabilitation and Applied Science
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• v.19 no.2
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• pp.87-96
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• 2003

The purposes of this study were to find out the optimum intensity of magnetic field where magnetism could promote the activity of osteoblast, and to discover the possibility of clinical application in the areas of dental implants and bone grafts by confirming the effect of clinically increasing bone formation. In this experiment, we used the Neodymium magnet, which had magnetic power six times as strong as the current ones and enabled the resistances against the demagnetization up to 20 to 50 times to be minimized with the size of 1mm in sight. In order to culture cells, a specially designed device was used. It was made to adjust the distance and accordingly to control the intensity of the magnetic field, by placing the cell culture plate in the center with a magnet of 1mm long and thick installed on the both ends. Using MC3T3-E1 cell, a kind of osteoblast-like cell, we cultured, for 24 hours, not only the test group which had been cultured under the magnetic fields with different intensity of 5, 10, 50, 100, 500, and 1000 Gauss, but also the control group excluding the influences of the magnetic field. After observing the cell's form and the density of the culture medium through an inverted microscope, we made a series of proceedings needed for the immunofluoroscence staining, such as fixation, normal serum reaction, primary antibody reaction, and secondary antibody reaction. And with a fluorescence microscope, we observed those-above and compared the frequency of expression of IFG-1 receptor. To make a Western immunoblotting analysis, the cells cultured under the same condition as the above had the procedure of the lysis buffer and the acrylamide gel electrophoresis was carried out. Protein transferred into the nitrocellulose membrane and tested on the primary and the secondary antibody reactions was observed and compared. The results were as follows: When observed through an inverted microscope, the nuclear divisions of the cells under the magnetic field of 10 Gauss were the most active, and the density of the cells could be observed the most enormously. As the result of an immunofluoroscence staining of IGF-1 receptor, the expression of IFG-1 was the most frequently observed under the magnetic field of 10 Gauss. On the other hand, few differences of consideration were made between the test group cultured under the magnetic fields of 5, 500, and 1000 Gauss and the control group. In respect of the expression of IFG-1 receptor, the test group cultured under the magnetic fields of 50 and 100 Gauss were higher than the control group, and lower than that cultured under the magnetic field of 10 Gauss.(p<0.05) According to the Western immunoblotting analysis, the band of IFG-1 receptor which had 85KDa of molecular weight was the darkest. Judging from the above-mentioned results, the growth factor receptor of an osteoblast cell which was an important criterion for the bone formation was increased in maximum under the magnetic field of 10 Gauss. Moreover it was observed that the optimum intensity of magnetic field in which magnetism made the activity of the osteoblast cell increase was about 10 Gauss.

• The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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• v.20 no.3
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• pp.131-140
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• 2015

To set up unicellular cyanobacterial strains with photo-biological $H_2$ production potential, live samples were repeatedly collected from 68 stations in the coastal zone of Korea for the four years since 2005. Among 77 cyanobacterial strains established six (KNU strains, CB-MAL002, 026, 031, 054, 055 and 058) were finally chosen as the excellent strains for $H_2$ production with $H_2$ accumulation over 0.15 mL $H_2\;mL^{-1}$ under general basic $H_2$ production conditions as well as positive $H_2$ production for more than 60 hr. To explore optimum procedures for higher $H_2$ production efficiency of the six cyanobacterial strains, the inter-strain differences in the growth rate under the gradients of water temperature and salinity were investigated. The maximum daily growth rates of the six strains ranged from 1.78 to 2.08, and all of them exhibited $N_2-fixation$ ability. Based on the similarity of the 16S rRNA sequences, all the test strains were quite close to Cyanothece sp. ATCC51142 (99%). The six strains, however, were grouped into separate clades from strain ATCC51142 in the molecular phylogeny diagram. Chlorophyll- a content was 3.4~7.8% of the total dried weight, and the phycoerythrin and phycocyanin contents were half of those in the Atlantic strain, Synechococcus sp. Miami BG03511. The growth of the six strains was significantly suppressed at temperatures above the optimal range, $30{\sim}35^{\circ}C$, to be nearly stopped at $40^{\circ}C$. The growth was not inhibited by high salinities of 30 psu salinity in all the strains while strain CB055 maintained its high growth rate at low salinities down to 15 psu. The euryhaline strains like CB055 might support massive biotechnological cultivation systems using natural basal seawater in temperate latitudes. base seawater. The biological and ecophysiological characteristics of the test strains may contribute to designing the optimal procedures for photo-biological $H_2$ production by unicellular cyanobacteria.

• Journal of Life Science
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• v.18 no.11
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• pp.1592-1599
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• 2008

The $\beta$-lactamase gene was cloned into E. coli DH5$\alpha$ from Bacillus sp. J105 with strong resistance against $\beta$-lactam antibiotics. The chromosomal DNA was partially digested with Sau3AI and ligated to BamHI digested pLAFR3. $\beta$-Lactamase positive clones were obtained by using in vitro packaging kit. The pKL11-${\Delta}4.6$ with $\beta$-lactamase activity was obtained by subcloning of the recombinant plasmid ($\beta$-lac +). The 6.5 kb fragment in the subcloned plasmid was sequenced. The DNA fragment that contains the $\beta$-lactamase gene encodes 309 amino acids. The 0.17 kb upstream region was similar to those of B. thuringinesis and B. cereus with 97% identity. The deduced amino acids sequence was also similar to those of $\beta$-lactamase from B. thuringinesis and B. cereus with 97% and 94% identity, respectively. The phylogenetic tree also showed the relationships of the $\beta$-lactamase gene of Bacillus sp. J105 to genetically related that of other Bacillus strains. Analysis of expression pattern of the pKL11-${\Delta}4.6$ in E. coli, revealed that the secretion efficiency of $\beta$-lactamase was $4{\sim}5%$ and the molecular weight was as same as that of original $\beta$-lactamase (31 kDa) from Bacillus sp. J105.

• Journal of Life Science
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• v.22 no.1
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• pp.98-109
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• 2012

The properties of lactate dehydrogenase (EC 1.1.1.27, LDH) and expression of monocarboxylate transporters (MCTs) 1, 2, and 4 were studied in tissues from Micropterus salmoides. Native-PAGE revealed that the LDH $A_4$ isozyme was predominantly located in skeletal muscle. The LDH $A_4$, $A_2B_2$, and $B_4$ isozymes were detected in heart, liver, eye, and brain tissues, while eye-specific $C_4$ isozyme was detected in eye tissue. In September, strong LDH $B_4$ isozyme activity was detected in heart tissue. High $A_4$ isozyme activity was noted in all other tissues except heart tissue. However, in November, strong $A_4$ isozyme activity was detected in heart tissue. The LDH/CS (Citrate synthase, EC 4.1.3.7) ratio in skeletal muscle and heart tissues indicated that anaerobic metabolism was high in those tissues. Native-PAGE after immunoprecipitation showed that eye-specific $C_4$ isozyme was more similar to the $A_4$ than the $B_4$ isozyme. The LDH $A_4$ isozyme was purified by affinity chromatography. The molecular weight of subunit A was 37,200. The LDH activity in tissues was consistently 11.05~28.32% due to inhibition by 10 mM pyruvate. The $K_m^{PYR}$ of LDH in eye tissue was very low. The optimum pH for LDH in tissues was pH 7.5~8.0. The LDH $A_4$ isozyme was detected in mitochondria of skeletal muscle, whereas the $B_4$ and $A_2B_2$ isozymes were detected in heart tissue mitochondria. Western blot analysis indicated that MCTs 1, 2, and 4 were located in the plasma membrane and mitochondria of skeletal muscle and heart tissues. The sizes of MCTs 1, 2, and 4 in skeletal muscle were 60, 54~38, and 63 kDa, while those in heart tissue were 57, 54~38, and 55.5 kDa, respectively. In conclusion, M. salmoides appears to use anaerobic metabolism predominantly when adapted to a hypoxic environment. In highly activated skeletal muscle and heart tissue, energy production is controlled by inward and outward flows of pyruvate and lactate through MCTs 1, 2, and 4 in the plasma membrane and mitochondria, with effective adjustment by LDH isozymes.

• Korean Journal of Ecology and Environment
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• v.33 no.4 s.92
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• pp.374-379
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• 2000

To elucidate the degradation mechanisms of organic materials in Lake Paldang, oxygen consuming rates in the water column, high molecular weight (HMW) organic materials and aggregates were analyzed. Also the chlorophyll a concentrations and environmental factors were measured three times in 1999. The concentrations of chlorophyll a ranged $5.5{\sim}14.2\;mg/m^3$ with the highest peak of $57.7\;mg/m^3$ at the surface water in April. Chlorophyll a concentration of aggregates retrieved from traps in 5 m and 20 m depths in May were 2779.5, $9044.8\;mg/m^3$, respectively. Those vlaues were more than 6 times higher compared with other months, and more than 49 times higher than water column chlorophyll a. Oxygen consuming rates of water column were in the narrow range of $0.4{\sim}0.5\;mg\;O_2\;l^{-1}\;day^{-1}$. HMW organic materials were using only small amount of oxygen, $0.01{\sim}0.04\;mg\;O_2\;l^{-1}\;day^{-1}$. The aggregates retrieved from 5 m depth by sediment trap consumed the oxygen in the range of $0.48{\sim}0.69\;mg\;O_2\;l^{-1}\;day^{-1}$, while aggregates collected from 20 m depth, 0.88 to $1.04\;mg\;O_2\;l^{-1}\;day^{-1}$. With these results, the HMW appeared not to be degraded in the water column, instead they seemed to be concentrated and affected the sediment oxygen demand.