• Korean Journal of Environmental Agriculture
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• v.41 no.2
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• pp.115-124
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• 2022

BACKGROUND: Methane is a major greenhouse gas attributed to global warming partly contributed by agricultural activities from ruminant fermentation and rice paddy fields. Methanotrophs are microorganisms that utilize methane. Their unique metabolic lifestyle is enabled by enzymes known as methane monooxygenases (MMOs) catalyzing the oxidation of methane to methanol. Rice absorbs, transports, and releases methane directly from soil water to its stems and the micropores and stomata of the plant epidermis. Methylobacterium species associated with rice are dependent on their host for metabolic substrates including methane. METHODS AND RESULTS: Methylobacterium spp. isolated from rice were evaluated for methane oxidation activities and screened for the presence of sMMO mmoC genes. Qualitatively, the soluble methane monooxygenase (sMMO) activities of the selected strains of Methylobacterium spp. were confirmed by the naphthalene oxidation assay. Quantitatively, the sMMO activity ranged from 41.3 to 159.4 nmol min^-1 mg of protein^-1. PCR-based amplification and sequencing confirmed the presence and identity of 314 bp size fragment of the mmoC gene showing over 97% similarity to the CBMB27 mmoC gene indicating that Methylobacterium strains belong to a similar group. CONCLUSION(S): Selected Methylobacterium spp. contained the sMMO mmoC gene and possessed methane oxidation activity. As the putative methane oxidizing strains were isolated from rice and have PGP properties, they could be used to simultaneously reduce paddy field methane emission and promote rice growth.

• KSBB Journal
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• v.8 no.4
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• pp.341-350
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• 1993

As an effort to develop an alternative transportation fuel, the production of methanol from methane gas was studied using the resting cells of an obligatory methanotroph, Methylosinus trichosporium OB3b. The reaction was carried out in high concentration phosphate buffer solutions with the flask-grown cells containing the exclusively cytoplasmic methane monooxygenase (sMMO) activity. The methanol accumulation rate was observed to be 79nmo1/mg·min during the initial 4.5hr. Phosphate-dependent inhibition was found for both sMMO and methanol dehydrogenase (MDH) activities, and the inhibition constants were 185mM and 42mM, respectively. The inhibition mode was noncompetitive. Methanol was found to be very inhibitory to the sMMO activity and the inhibition constant (noncompetitive) was 21mM when propylene was used as substrate. The sMO activity in the resting cells was declined very fast and the rate became very high during the methanol production. These results indicate that the use of M. trichosporium OB3b as a biocatalyst for the methanol production is heavily dependent on the stable maintenance of the whole-cell SMO activity as well as the effective alleviation of product inhibition.

• KSBB Journal
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• v.13 no.5
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• pp.483-490
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• 1998

Two unidentified methanotrophic strains (MM-white and MM-red) secreting soluble methane monooxygenase (sMMO) involved in thrichloroethylene biodegradation have been isolated from mixed methanotrophic consortium (MM) around Taejon area. Subsequently four methanotrophic strains were isolated from MM and named according to their color; white (MS-white), yellow (MS-yellow), pink (MS-pink) and reddish brown (MS-rbrown). All strains except MS-yellow which can take glucose as well as methane, metabolized methane as a sole carbon source. They all showed symbiotic behavior when methane was used as the sole carbon source. Optimum conditions of cell growth for MM were pH of 6.8 - 7.2, temperature of 29 - 32$^{\circ}C$, and gas flow rate of 6 (for methane), 40 (for air), and 4 ml/min (for carbon dioxide). The sMMO activity was expressed as naphthalene oxidation rate (${\mu}$mol/ mg protein/ hr). The sMMO activity for MM grown in flask culture with 1 ${\mu}$M of CuSO4 was 36, while it was 61 without copper. The activity for MM grown in the fermentor without CuSO4 was 1077, but is was 197 after reaction with 5 ppm of TCE. The methanotrophs showed significantly high sMMO activity despite the presence of 1 ${\mu}$M of CuSO4, although most of other strains already known could not express sMMO activity under this condition.

• Journal of the Korea Institute of Information and Communication Engineering
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• v.16 no.11
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• pp.2563-2568
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• 2012

The growth speed of the industry of national smart phone game service has become faster and the its economical and cultural effect has increased steadily these days. The DB design for data perpetuation in C/S MMO which has accumulated data a lot and many simultaneous plays like smart phone games is very important. This paper, mindful of the operation and the scalability suitable for android OS, by utilizing RDBMS, enabling very short, many transactions, their complexity, and a large amount of game data process, suggests fully-qualified MMORPG game service architecture design techniques widely available for more than online game industries.

• Proceedings of the Korean Institute of Information and Commucation Sciences Conference
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• 2012.10a
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• pp.465-468
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• 2012

The growth speed of the industry of national smart phone game service has become faster and the its economical and cultural effect has increased steadily these days. The DB design for data perpetuation in C/S MMO which has accumulated data a lot and many simultaneous plays like smart phone games is very important. This paper, mindful of the operation and the scalability suitable for android OS, by utilizing RDBMS, enabling very short, many transactions, their complexity, and a large amount of game data process, suggests fully-qualified MMORPG game DB design techniques widely availabe for more than online game industries.

• Biotechnology and Bioprocess Engineering:BBE
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• v.8 no.5
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• pp.309-312
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• 2003

The transformation capacity (T$\_$c/) of Methylosinus trichosporium OB3b in the degradation of ethylene chlorides was determined by measuring the decrease of soluble methane monooxygenase (sMMO) activity of resting cells in batch experiments. All measurements of sMMO activity were taken in the presence of 20 mM formate to avoid the deficiency of reducing power, and within 2 hrs to avoid the effect of natural inactivation from instability of the resting cells. The constant T$\_$c/ values of 0.58 ${\pm}$ 0.132 and 0.80 ${\pm}$ 0.17 ${\mu}$mol/mg cell were obtained for trichloroethylene (TCE) and 1,2-dichloroethylene (cis and trans-1,2-DCE), respectively, regardless of their concentrations. The transformation capacity measured by this method can be used to predict the amount of cells that should be stimulated in in-situ bioremediation.

• Journal of The Korean Society of Grassland and Forage Science
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• v.20 no.4
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• pp.243-250
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• 2000

Selection of oat variety according to cultivation period is an important to increase productivity of forage crop. To investigate the growth characters and dry matter productivity of oats according to cultivation period, early-maturing oat(EM0) and medium-maturing oat(MM0) variety were sown in spring and autumn at central district of Korea. When oats were cultivated in spring, MMO variety was longer 8 cm in leaf length and wider 5 mm in leaf width than 25 cm and 12 mm of EM0 variety, rexpectively. but in autumn, those were not different between maturities of oat varieties. Plant length of MMO variety was longer 7 cm than that of EM0 Variety in spring cultivation, but in autumn, plant length of EM0 variety was longer 14 cm than that of MMO variety. Heading dates were 20th to 22th May in EM0 variety, and 30th May to 10th June in MMO variety in spring cultivation, and ear of EM0 variety emerged 30th Oct. to loth Nov. but that of MMO variety did not emerge at harvesting day in autumn cultivation. In Spring cultivation, dry matter(DM) yield of EM0 variety was 7,349 kg/ha that was less 56% than 11,466 kg/ha of MMO variety, but In Autumn, was more 15% than 6,575 kg/ha of MMO variety. In conclusion, selection of oat variety in spring and autumn cultivation can be different with cropping system of forage crop, and this trial is suggesting that oat variety could be sown MMO variety in spring and EM0 variety in autumn at central district of Korea. (Key words : Oat variety, Productivity, Cultivation season)

• KSBB Journal
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• v.14 no.4
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• pp.452-459
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• 1999

A two-stage continuous stirred tank reactor (CSTR)/trickling biofilter reactor (TBR) system was developed for the degradation of gas-phase trichloroethlene (TCE) using Methylosinus trichoporium OB3b. Mrthylosinus trichosporium OB3b was immobilized on activated carbons in TBR and the microbial growth reactor of a CSTR was coupled for the reactivation of the deactivated cells during TCE degradation. The effect of operation variables on TCE conversion and degradation rate were studied. At inlet TCE concentrations ranging from 10 to 80 $\mu$mol/L, TCE degradation rate was increased up to 525 mg TCE/Lㆍday with 75% conversion. The TCE degradation rates were also increased with increse in broth recycle flow rate, gas flow rate and dilution rate. When the temperature of TBR was changed from 3$0^{\circ}C$ to 15$^{\circ}C$, TCE degradation rate and TCE conversion were increased due to the enhanced TCE transfer from gas-phase. The two-stage reactor system was found to be stable and has been operated for more than 270 days.

• Journal of Nutrition and Health
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• v.41 no.8
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• pp.701-710
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• 2008

The wild simulated ginseng (WSG) has been effectively used in folk medicine as a remedy against hepatic disease, hypertension and arthritic disease. However, there is still lack of scientific proof about its antioxidant capability. The present study has been conducted to evaluate the protective role of the WSG ethanol extract in the CCl4-induced oxidative stress and resultant hepatic disfunction in ICR mice. The electron donating abilities and IC50 of WSG etnanol extract were 76.86 ${\pm}$ 1.06% and 33.3 ${\mu}g$/mL (that of ascobic acid was 16.5 ${\mu}g$/mL), respectively. Total antioxidant status of WSG extract was 2.13 ${\pm}$ 0.06 mmoL/mg, while the values of ascorbic acid and BHT were 3.63 ${\pm}$ 0.06 and 3.12 ${\pm}$ 0.02, respectively. ICR mice (aged 3weeks) were fed for 4 weeks on AIN-93M diet and had free access to food and water. The animals were divided into three groups: normal group (intraperitoneally (i.p) injected with PBS at 100 ${\mu}L$/mouse), group C; CCl4-induced and without any treatment. (i.p injected only PBS, 100 ${\mu}L$ /mice), group G; CCl4-induced and treated with WSG (i.p injected with 5 mg WSG extract per mouse, suspended in 100 ${\mu}L$ phosphate buffer). After the i.p. injection of WSG or PBS (5 times for 7weeks), all mice were administered CCl4 in olive oil at the last day of the experiment, except for normal group. The normal group was administered only olive oil. Determination of plasma triglyceride, total cholersterol, fasting glucose and GPT activity was performed using automatic blood analyzer. To evaluate the protective effect against the oxidative stress, DNA fragmentation and TBARS were determined in blood leucocytes and RBC and hepatocyte, respectively. Body and organs weights and food intake did not show significant differences among the groups. Blood total cholesterol of group G was similar to that of normal group, which was the lowest in group C. The fasting blood glucose level was the highest in normal group (205.20 ${\pm}$ 135.24), which were decreased in group C (134.2 ${\pm}$ 79.31) and group G (126.48 ${\pm}$ 77.05). TBARS values in a red blood cell and hepatic tisuue homogenate were lower in group G comparing to the group C. DNA% in tail, tail length (TL) and tail moment (TM) of blood leucoocytes showed the highest values in group C (20.11 ${\pm}$ 2.47, 17.36 ${\pm}$ 2.58, 94.11 ${\pm}$ 12.29) and they were significantly diminished in group G (9.63 ${\pm}$ 1.19, 7.04 ${\pm}$ 1.50, 38.64 ${\pm}$ 7.60). In conclusion, wild simulated ginseng might be a protective agent against the oxidative stress.

• 한국생물공학회:학술대회논문집
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• 2000.04a
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• pp.485-488
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• 2000

Methyl mercaptan oxidase was isolated and purified from Thiobacillus thioparus TK-m for the detection of mercaptans. The procedure of purification involved DEAE-Sephacel and Superose 12 column chromatographies with recovery yields of 47.5 and 48.5 %, and specific activity of 374 and 1240.8 units/mg-protein, respectively. The molecular weight of purified methyl mercaptan oxidase was determined to be 66.1 kDa by SDS-PAGE. Optimum temperature for activity was observed at $55\;^{circ}C$. This enzyme was activated by $(NH_4)_2SO_4$ and NaCl and inhibited by $NH_4Cl$.