• BMB Reports
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• v.35 no.3
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• pp.330-336
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• 2002

The lgtB genes that encode $\beta$-1,4-galactosyltransferases from Neisseria meningitidis ATCC 13102 and gonorrhoeae ATCC 31151 were isolated by a polymerase chain reaction using the pfu DNA polymerase. They were expressed under the control of lac and T7 promoters in Escherichia coli M15 and BL21 (DE3). Although the genes were efficiently expressed in E. coli M15 at $37^{\circ}C$ (33 kDa), most of the $\beta$-1,4-galactosyltransferases that were produced were insoluble and proteolysed into enzymatically inactive polypeptides that lacked C-terminal residues (29.5 kDa and 28 kDa) during the purification steps. When the temperature of the cell growth was lowered to $25^{\circ}C$, however, the solubility of the $\beta$-1,4-galactosyltransferases increased substantially. A stable N-terminal his-tagged recombinant enzyme preparation could be achieved with E. coli BL21 (DE3) that expressed lgtB. Therefore, the cloned $\beta$-1,4-galactosyltransferases were expressed under the control of the T7 promoter in E. coli BL21 (DE3), mostly to the soluble form at $25^{\circ}C$. The proteins were easily purified to homogeneity by column chromatography using Ni-NTA resin, and were found to be active. The galactosyltransferases exhibited pH optimum at 6.5-7.0, and had an essential requirement for the $Mn^{+2}$ ions for its action. The $Mg^{+2}$ and $Ca{+2}$ ions showed about half of the galactosyltransferase activities with the $Mn^{+2}$ ion. In the presence of the $Fe^{+2}$ ion, partial activation was observed with the $\beta$-1,4-galactosyltransferase from N. meningitidis(64% of the enzyme activity with the $Mn^{+2}$$Ni^{+2}$, $Zn^{+2}$, and $Cu^{+2}$ ions could not activate the $\beta$-1,4-galactosyltransferase activity. The inhibited enzyme activity with the $Ni^{+2}$ ion was partially recovered with the $Mn^{+2}$$Fe^{+2}$, $Zn^{+2}$, and $Cu^{+2}$ ions, the $Mn^{+2}$$\beta$-1,4-galactosyltransferase activity was 1.5-fold stimulated with the non-ionic detergent Triton X-100 (0.1-5%).

• Microbiology and Biotechnology Letters
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• v.9 no.3
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• pp.159-164
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• 1981

Reinforced Calcium-alginate gel entrappment method for enzyme immobilization is described with an example of penicillin amidase from Bacillus megaterium KFCC 10029, a partially constitutive mutant of B. megaterium ATCC 14945. Penicillin amidase recovered from the fermentation broth by adsorption on celite is mixed with alginate and gelatin solution, and cast into a pellet or noodle form by coagulation in calcium salt solution followed by crosslinking with glutaraldehyde. Optimum pH and temperature of the immobilized enzyme preparation were 8.0 and 6$0^{\circ}C$, respectively. Kinetic constants such as Km value and the inhibition constant of 6-APA and phenylacetic acid were 2.6 mM, 7.4 mM and 21.2 mM, respectively. The enzyme leakage from the adsorbent during operation was successfully prevented owing to the increase of physical strength of gel coat. The half lives in a column reactor were 6 and 30 days at the respective temperature of 4$0^{\circ}C$ and 3$0^{\circ}C$, which were the 6-8 fold increased values as compared with those of without entrappment. The results highly recommended the use of reinforced Calcium-alginate gel entrappment method for the enhancement of physical strength and the operational stability of alginate gel entrapped enzyme.

• The Korean Journal of Mycology
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• v.20 no.4
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• pp.324-336
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• 1992

On the five interspecific protoplast fusants of Ganoderma lucidum and G. applanatum was the antitumor test performed. The fusant F-2 was selected, to examine the cultured mycelia (protein bound polysaccharide) as antitumor components. When a dose of 20 mg/kg/day of each components purifed from F-2 fusant was, i.p., injected into ICR mice, the inhibition ratio of Fr. II against the solid form of sarcoma 180 increased to 1.5 times as compared with that of their parents. When Fr. II was examined for immunopotentiation activity, it increased the amount of the superoxide anion in activated macrophages to 1.2 times and the count of hemolytic plaque forming cells in the spleen to 4.3 times as compared with that of each control group. Its chemical analysis showed 85.2% polysaccharide which consisted of glucose, galactose, mannose, fucose and xylose, and 0.39% protein of 15 amino acids. The content of hexosamine was 0.39% and the molecular weight of Fr. V was $5.6{\times}10^4$ dalton.

• Journal of Korean Society for Atmospheric Environment
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• v.34 no.4
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• pp.554-566
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• 2018

The rice husk contains nutrients which can be easily utilized by microorganisms, and also has a water retaining ability, which played a crucial part in enabling it to become a biofilter media. In this study, we evaluated the applicability of rice husk pellet bio-scrubber as a microbiological carrier. The pelletization experiment of rice husk as a biological media was performed using PVA and EVA binder. Also, the feasibility tests of rice husk as a biological media for odor removal were carried out in order to know whether rice-husk contains useful components as a media for microbiological growth or not. Lastly, a combined test for odor gas absorption and biological oxidation was conducted using a lab scale bio-filter set-up packed with rice-husk pellets as wet-scrubber. The major components of the rice husk were carbon, hydrogen, nitrogen, and oxygen, while carbon acted as the main ingredient which comprised up to 23.00%. The C : N : P ratio was calculated as 45 : 1 : 2. Oxygen uptake rate, yield and decay rate of the rice husk eluent was calculated to be $0.0049mgO_2/L/sec$, 0.24 mgSS/mgCOD and 0.004 respectively. The most stable form of rice husk pellets was produced when the weight of the rice husk, EVAc, PVAc, and distilled water was 10 : 2 : 0.2 : 10. The prepared rice husk pellets had an apparent density of 368 g/L and a porosity of 59.00% upon filling. Dry rice husks showed high adsorption capacity for ammonia gas but low adsorption capacity for hydrogen sulfide. The bio-filter odor removal column filled with rice husk pellets showed more than 99.50% removal efficiency for NH3 and H2S gas. Through the analysis of circulation water, the prime removal mechanism is assumed to be the dissolution by water, microbial nitrification, and sulfation. Finally, it was confirmed that the microorganisms could survive well on the rice husk pellets, which provided them a stable supply of nutrients for their activity in this long-term experiment. This adequate supply of nutrients from the rice husk enabled high removal efficiency by the microorganisms.

• Journal of the Korea Concrete Institute
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• v.24 no.3
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• pp.259-266
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• 2012

A general earthquake resistant design philosophy of ductile frame buildings allows beams to form plastic hinges adjacent to beam-column connections. In order to carry out this design philosophy, the ultimate bond or shear strength of the beam should be greater than the flexural yielding force and should not degrade before reaching its required ductility. The behavior of RC members dominated by bond or shear action reveals a dramatic reduction of energy dissipation in the hysteretic response due to the severe pinching effects. In this study, a method was proposed to predict the deformability of reinforced concrete members with short-span-to-depth-ratios, which would result in bond failure after flexural yielding. Repeated or cyclic loading produces a progressive deterioration of bond that may lead to failure at lower cyclic bond stress levels. Accumulation of bond damage is caused by the propagation of micro-cracks and progressive crushing of concrete in front of the lugs. The proposed method takes into account bond deterioration due to the degradation of concrete in the post yield range. In order to verify bond deformability of the proposed method, the predicted results were compared with the experimental results of RC members reported in the technical literature. Comparisons between the observed and calculated bond deformability of the tested RC members showed reasonably good agreement.

• Journal of the Korean Society of Food Science and Nutrition
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• v.16 no.3
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• pp.75-84
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• 1987

Lipids in Bush Clover (Lespedza bicolor) seed were extracted with the mix ture of chloro-form-methanol (2 : 1, v/v) and then fractionated into neutral lipids, glycolipids and phospholipids by silicic acid column chromatography. Components and fatty acid composition of each fraction were determined by thin layer and gas chromatographies. The results were summarized as follows. In Bush Clover seed, the contents of neutral lipids, glycolipids and phospholipids were 71.75%, 23.26% and 4.99% respectively. Triglycerides(61.90%) and free fatty acids(22.04%) were the major components among the neutral lipids. Esterified sterols, free sterols, diglycerides and monoglycerides were the minor components. The major components of glycolipids were monogalactosyl diglycerides(38.19%) the others were esterified steryl glycosides, cerebrosides and digalactosyl diglycerides. The major components of the phospholipids were phosphatidyl cholines(36.46%), phosphatidyl inositols(21.52%) and phosphatidyl ethanolamines(17.29%). The major fatty acid of total lipid, neutral lipids and glycolipids were linoleic acid, linolenic acid, oleic acid and palmitic acid. On the other hand, predominate fatty acid of phospholipids were linoleic acid, palmitic acid, linolenic acid and stearic acid.

• Journal of Microbiology and Biotechnology
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• v.16 no.10
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• pp.1583-1590
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• 2006

T4 endonuclease V (T4 endo V) [EC 3. 1. 25. 1], found in bacteriophage T4, is responsible for excision repair of damaged DNA. The enzyme possesses two activities: a cyclobutane pyrimidine dimer DNA glycosylase (CPD glycosylase) and an apyrimidic/apurinic endonuclease (AP lyase). T4 denV (414 bp cDNA) encoding T4 en do V (138 amino acid) was synthesized and expressed using either an expression vector, pTriEx-4, in E. coli or a baculovirus AcNPV vector, pBacPAK8, in insect cells. The recombinant His-Tag/T4 endo V (rHis-Tag/T4 endo V) protein expressed from bacteria was purified using one-step affinity chromatography with a HiTrap Chelating HP column and used to make rabbit anti-His-Tag/T4 endo V polyclonal antibody for detection of recombinant T4 endo V (rT4 endo V) expressed in insect cells. In the meantime, the recombinant baculovirus was obtained by cotransfection of BacPAK6 viral DNA and pBP/T4 endo V in Spodoptera frugiperda (Sf21) insect cells, and used to infect Sf21 cells to overexpress T4 endo V protein. The level of rT4 endo V protein expressed in Sf21 cells was optimized by varying the virus titers and time course of infection. The optimal expression condition was set as follows; infection of the cells at a MOI of 10 and harvest at 96 h post-infection. Under these conditions, we estimated the amount of rT4 endo V produced in the baculovirus expression vector system to be 125 mg/l. The rT4 endo V was purified to homogeneity by a rapid procedure, consisting of ion-exchange, affinity, and reversed phase chromatographies, based on FPLC. The rT4 endo V positively reacted to an antiserum made against rHis-Tag/T4 endo V and showed a residual nicking activity against CPD-containing DNA caused by UV. This is the first report to have T4 endo V expressed in an insect system to exclude the toxic effect of a bacterial expression system, retaining enzymatic activity.

• Journal of Biosystems Engineering
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• v.25 no.3
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• pp.221-226
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• 2000

Hot air heater with light oil burner is the most common heater for greenhouse heating in the winter season in Korea. However, since the thermal efficiency of the heater is about 80∼85%, considerable unused heat amount in the form of exhaust gas heat discharges to atmosphere. In order to capture this exhaust heat a heat recovery system for plant bed heating in the greenhouse was built and tested in the hot air heating system of greenhouse. The heat recovery system is made for plant bed or soil heating in the greenhouse. The system consisted of a heat exchanger made of copper pipes, ${\Phi}12.7{\times}0.7t$ located in the rectangular column of $330{\times}330{\times}900mm$, a water circulation pump, circulation plastic pipe and a water tank. The total heat exchanger area is 1.5$m^2$, calculated considering the heat exchange amount between flue gas and water circulated in the copper pipes. The system was attached to the exhaust gas path. The heat recovery system was designed as to even recapture the latent heat of flue gas when exposing to low temperature water in the heat exchanger. According to the performance test it could recover 45,200 to 51,000kJ/hr depending on the water circulation rates of 330 to $690\ell$/hr from the waste heat discharged. The exhaust gas temperature left the heat exchanger dropped to $100^{\circ}C$ from $270^{\circ}C$ by the heat exchange between the water and the flue gas, while water gained the difference and temperature increased to $38^{\circ}C$ from $21^{\circ}C$ at the water flow rate of $690\ell$/hr. By the feasibility test conducted in the greenhouse, the system did not encounter any difficulty in operations. And, the system could recover 220,235kJ of exhaust gas heat in a day, which is equivalent of 34% of the fuel consumption by the water boiler for plant bed heating of 0.2ha in the greenhouse.

• Journal of agriculture & life science
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• v.45 no.6
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• pp.251-263
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• 2011

In order to investigate treatment effects of limestone and steel refining slag for paddy soils contaminated with arsenic and heavy metals, a lab-column test was carried out under reducing environments of flooded paddy soils. In conditions of the flooded paddy soils, at the point of time when iron and manganese were reduced and leached rapidly, heavy metals also leached rapidly, and some leachate samples from an untreated soil exceeded regulatory standards. On the contrary, all samples from soils treated with limestone 5% and steel refining slag 5% respectively were below the regulatory standards, showing much lower heavy metal concentrations than in the untreated soil. Arsenic increased continuously during the observation period according to its typical characteristics, and along with decreasing redox potential, arsenic was expected to leach as $H_3AsO_3$-of form $A^{3+}$ with high mobility and strong toxicity. Limestone and steel refining slag showed high treatment effects against heavy metals present in soil and steel refining slag especially showed the high treatment effects against arsenic.

• Journal of the Korean Geosynthetics Society
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• v.20 no.4
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• pp.43-52
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• 2021

As port development in soft ground is actively promoted for international logistics and transportation, the Deep Mixing Method (DMM) is continuously applied to form an improved column body directly in the ground by mixing cement with soil to secure the stability of the structure. However, in the case of cement, there is a problem of emitting a lot of greenhouse gases during the production process, so the development and use of new alternative materials are socially required to achieve the national goal of carbon neutrality. Accordingly, in this study, CMD-SOIL, developed to induce a hardening reaction similar to cement by recycling recycled resources, was used as a ground improvement material for the DMM. In addition, it was attempted to determine the possibility of replacing cement by conducting on-site test construction and evaluating applicability. As a result of the study, the compressive strength of CMD-SOIL compared to the design reference strength was 1.46 to 2.64 times higher in the field mixing test and 1.2 to 5.03 times higher than in the confirmed boring. In addition, the ratio (λ) of the compressive strength in the field to the design reference strength was 0.63 to 1.14, which was similar to the previous research results. Therefore, in the case of CMD-SOIL, it is possible to express the compressive strength necessary to secure stability, and there is no difference in applicability compared to existing materials such as ordinary portland cement and blast furnace slag cement, so it was analyzed that it could be used as a ground improvement material for the DMM.