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Development of the feedback resistant pheAFBR from E. coli and studies on its biochemical characteristics (E. coli 유래 pheA 유전자의 되먹임제어 저항성 돌연변이의 구축과 그 단백질의 생화학적 특성 연구)

  • Cao, Thinh-Phat;Lee, Sang-Hyun;Hong, KwangWon;Lee, Sung Haeng
    • Korean Journal of Microbiology
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    • v.52 no.3
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    • pp.278-285
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    • 2016
  • The bifunctional PheA protein, having chorismate mutase and prephenate dehydratase (CMPD) activities, is one of the key regulatory enzymes in the aromatic amino acid biosynthesis in Escherichia coli, and is negatively regulated by an end-product, phenyalanine. Therefore, PheA protein has been thought as useful for protein engineering to utilize mass production of essential amino acid phenylalanine. To obtain feedback resistant PheA protein against phenylalanine, we mutated by using random mutagenesis, extensively screened, and obtained $pheA^{FBR}$ gene encoding a feedback resistant PheA protein. The mutant PheA protein contains substitution of Leu to Phe at the position of 118, displaying that higher affinity (about $290{\mu}M$) for prephenate in comparison with that (about $850{\mu}M$) of wild type PheA protein. Kinetic analysis showed that the saturation curve of $PheA^{FBR}$ against phenyalanine is hyperbolic rather than that of $PheA^{WT}$, which is sigmoidal, indicating that the L118F mutant enzyme has no cooperative effects in prephenate binding in the presence of phenylalanine. In vitro enzymatic assay showed that the mutant protein exhibited increased activity by above 3.5 folds compared to the wild type enzyme. Moreover, L118F mutant protein appeared insensitive to feedback inhibition with keeping 40% of enzymatic activity even in the presence of 10 mM phenylalanine at which the activity of wild type $PheA^{WT}$ was not observed. The substitution of Leu to Phe in CMPD may induce significant conformational change for this enzyme to acquire feedback resistance to end-product of the pathway by modulating kinetic properties.

Characteristics of Lactate Dehydrogenase Produced from Lactobacillus sp. FFy111-1 as a Ruminant Probiotic (반추동물용 활성제로서 Lactobacillus sp. FFy111-1이 생산한 Lactate Dehydrogenase의 특성에 관한 연구)

  • Sung, H.G.;Kim, D.K.;Bae, H.D.;Shin, H.T
    • Journal of Animal Science and Technology
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    • v.46 no.4
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    • pp.625-634
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    • 2004
  • The objective of this experiment is to study the possibility of lactate dehydrogenase(LDH) enzyme to prevent lactate accumulation in the rumen, For understanding capacity of bacterial LDH in rumen environments, this study was conducted to explore the effects of temperature, pH, VFAs and metal ions on Lactobacillus sp. FFy111-1's LDH activity, and the LDH activation in rumen fluid accumulated lactate. The optimum pH and temperature of LDH were pH 7.5 and 40$^{\circ}C$, respectively. The LDH activity had a good thennostability at range from 30 to 50$^{\circ}C$. The highest pH stability of the enzyme was at ranges from pH 7.0 to 8.0 and the enzyme activities showed above 64% level of non-treated one at pH 6.0 and 6.5. The LDH was inactivated by VFAs treatments but was enhanced by metal ion treatments without NaCl and $CuSO_4$ Especially, the LDH activity was increased to 127% and 124% of its original activity by 2 mM of $BaCl_2$ and $MnSO_4$, addition, respectively. When the acidic rumen fluid was treated by LDH enzyme of Lactobacillus sp. FFy111-1, the lactate concentration in the rumen fluid was lower compared with non-treated rumen fluid(P<0.05). This lactate reduction was resulted from an action of LDH. It was proved by result of purified D,L-LDH addition that showed the lowest lactate concentration among the treatments(P<0.05). Although further investigation of microbial LDH and ruminal lactate is needed, these findings suggest that the bacterial LDH has the potential capability to decrease the lactate accumulated in an acidic rumen fluid. Also, screening of super LDH producing bacteria and technical development for improving enzyme activity in rumen environment are essential keys for practical application.

Agricultural Geography of Rice Culture in California (미국 캘리포니아주(州)의 벼농사에 관한 농업지리학적 연구)

  • Lee, Jeon;Huh, Moo-Yul
    • Journal of the Korean association of regional geographers
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    • v.2 no.1
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    • pp.51-67
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    • 1996
  • There are three main rice-growing regions in the United States: the prairie region along the Mississippi River Valley in eastern Arkansas; the Gulf Coast prairie region in southwestern Louisiana and southeastern Texas; and the Central Valley of California. The Central Valley of California is producing about 23% of the US rice(Fig. 1). In California. most of the crop has been produced in the Colusa, Sutter, Butte, Glenn Counties of the Sacramento Valley since 1912, when rice was commercially grown for the first time in the state(Fig. 2). Roughly speaking, the average annual area sown to rice in California is about 300,000 acres to 400,000 acres during the last forty years(Fig. 3). California rice is grown under a Mediterranean climate characterized by warm, dry, clear days, and a long growing season favorable to high photosynthetic rates and high rice yields. The average rice yield per acre is probably higher in California than in any other rice-growing regions of the world(Fig. 4). A dependable supply of irrigation water must be available for a successful rice culture. Most of the irrigation water for California rice comes from the winter rain and snow-fed reservoir of the Sierra Nevada mountain ranges. Less than 10 percent of rice irrigation water is pumped from wells in areas where surface water is not sufficient. It is also essential to have good surface drainage if maximum yields are to be produced. Rice production in California is highly mechanized, requiring only about four hours of labor per acre. Mechanization of rice culture in California includes laser-leveler technology, large tractors, self-propelled combines for harvesting, and aircraft for seeding, pest control, and some fertilization. The principal varieties grown in California are medium-grain japonica types with origins from the cooler rice climates of the northern latitudes (Table 1). Long-grain varieties grown in the American South are not well adapted to California's cooler environment. Nearly all the rice grown recently in California are improved into semidwarf varieties. Choice of variety depends on environment, planting date, quality desired, marketing, and harvesting scheduling. The Rice Experiment Station at Biggs is owned, financed, and administered by the rice industry. The station was established in 1912, as a direct result of the foresight and effort of Charles Edward Chambliss of the United States Department of Agriculture. Now, The station's major effort is the development of improved rice varieties for California.

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The Cross-sectional Mass Flux Observation at Yeomha Channel, Gyeonggi Bay at Spring Tide During Dry and Flood Season (단면 관측을 통한 경기만 염하수로의 대조기 평수시와 홍수시 유출입량 변화특성 조사)

  • Lee, Dong-Hwan;Yoon, Byung-Il;Kim, Jong-Wook;Gu, Bon-Ho;Woo, Seung-Buhm
    • Journal of Korean Society of Coastal and Ocean Engineers
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    • v.24 no.1
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    • pp.16-25
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    • 2012
  • To calculate the total mass flux that change in dry and flood season in the Yeomha Channel of Gyeonggi Bay, the 13 hour bottom tracking observation was performed from the southern extremity. The value of the total mass flux(Lagrange flux) was calculated as the sum of the Eulerian flux value and stroke drift value and the tidal residual flow was harmonically analyzed through the least-squares method. Moreover, the average during the tidal cycle is essential to calculate the mass flux and the tidal residual flow and there is the need to equate the grid of repeatedly observed data. Nevertheless, due to the great differences in the studied region, the number of vertical grid tends to change according to time and since the horizontal grid differs according to the transport speed of the ship as a characteristic of the bottom tracking observation, differences occur in the horizontal and vertical grid for each hour. Hence, the present study has vertically and horizontally normalized(sigma coordinate) to equate the grid per each hour. When compared to the z-level coordinate system, the Sigma coordinate system was evaluated to have no irrationalities in data analysis with 5% of error. As a result of the analysis, the tidal residual flow displayed the flow pattern of sagging in the both ends in the main waterway direction of dry season. During flood season, it was confirmed that the tidal residual flow was vertical 2-layer flow. As a result of the total mass flux, the ebb properties of 359 cm/s and 261 cm/s were observed during dry and flood season, respectively. The total mass flux was moving the intertidal region between Youngjong-do and Ganghwa-do.

Changes in Functional Compounds and Antioxidant Activities in Storage Duration with Accelerated Age-Conditioning of Oats (노화가속화 조건에서 저장 기간에 따른 귀리의 기능성 성분 및 항산화 활성 변화)

  • Son, Yu Rim;Lee, Ji Hae;Park, Hyoung-Ho;Lee, Byong Won;Kim, Hyun-Joo;Han, Sang-Ik;Woo, Koan Sik;Lee, Byoung-kyu;Lee, Sang-Chul;Lee, Yu Young
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.63 no.2
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    • pp.149-157
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    • 2018
  • In this study, we investigated the changes in physicochemical properties, antioxidant activities, and contents of functional compounds, such as avenanthramides (AVNs), vitamin E, and ${\beta}$-glucan, in oats by accelerated age-conditioning (temperature: $45^{\circ}C$, relative humidity: 20%). No significant differences were observed in crude protein, crude fat, and AVNs contents of three oat cultivars, up to 63 days of storage; however, their antioxidant activities, as well as ${\beta}$-glucan, vitamin E, and fatty acid contents were significantly different (p < 0.05). ${\beta}$-glucan and fatty acid contents and the antioxidant activities of Deayang (DY) cultivar did not change during storage. ${\beta}$-glucan and unsaturated fatty acid contents of Choyang (CY) and Jopung (JP) increased during the storage period, while antioxidant activities did not (DPPH-CY; 48.1 to 26.9 mg TEAC/100 g, JP; 49.4 to 26.7 mg TEAC/100 g. ABTS-CY; 88.4 to 56.3 mg TEAC/100 g, JP; 80.0 to 55.8 mg TEAC/100 g). The total vitamin E content in DY (1.20 to 0.85 mg/100 g) and CY (1.73 to 1.33 mg/100 g) decreased, but it was maintained in JP. This study indicated that the changes in physicochemical properties and functional compounds of oat grains during storage depends on the cultivars. The result showed that DY, which has the highest AVNs content, has more stable functional compounds and antioxidant activities during storage. These results can serve as essential data for post-harvest management and development of functional food materials for extending the use of oats.

Neural and Cholinergic Differentiation of Mesenchymal Stem Cells Derived from the Human Umbilical Cord Blood (인간 제대혈액에서 유래된 중간엽 줄기세포의 신경 및 콜린성 분화)

  • Kam, Kyung-Yoon;Kang, Ji-Hye;Do, Byung-Rok;Kim, Hea-Kwon;Kang, Sung-Goo
    • Development and Reproduction
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    • v.11 no.3
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    • pp.235-243
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    • 2007
  • Human umbilical cord blood(HUCB) contains a rich source of hematopoietic stem cells, mesenchymal stem cells and endothelial cell precursors. Mesenchymal stem cells(MSCs) in HUCB are multipotent stem cells, differ from hematopoietic stem cells and can be differentiated into neural cells. We studied on transdifferentiation-promoting conditions in neural cells and cholinergic neuron induction of HUCB-derived MSCs. Neural differentiation was induced by addingdimethyl sulphoxide(DMSO) and butylated hydroxyanisole(BHA) in Dulbeco's Modified Essential Medium(DMEM) and fetal bovine serum(FBS). Differentiation of MSCs to cholinergic neurons was induced by combined treatment with basic fibroblast growth factor(bFGF), retinoic acid(RA) and sonic hedgehog(Shh). MSCs treated with DMSO and BHA rapidly assumed the morphology of multipolar neurons. Both immunocytochemistry and RT-PCR analysis indicated that the expression of a number of neural markers including $\beta$-tubulin III, GFAP and MBP, was markedly elevated during this acute differentiation. The differentiation rate was about $32.3{\pm}2.9%$ for $\beta$-tubulin III-positive cells, $11.0{\pm}0.9%$ for GFAP, and $9.4{\pm}1.0%$ for Gal-C. HUCB-MSCs treated combinatorially with bFGF, RA and Shh were differentiated into cholinergic neurons. After cholinergic neuronal differentiation, the $\beta$-tubulin III-positive cell population of total cells was $31.3{\pm}3.2%$ and of differentiated neuronal population, $70.0{\pm}7.8%$ was ChAT-positive showing 3 folds higher in cholinergic population than neural induction. Conclusively, HUCB-derived MSCs can be differentiated into neural and cholinergic neurons and these findings suggest that HUCB are alternative cell source of treatment for neurodegenerative diseases such as Alzheimer's disease.

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Shipborne Mobile LiDAR(Light Detection and Ranging) System for the Monitoring of Coastal Changes (해안지형 모니터링을 위한 해상모바일라이다 지형 측정 시스템 구축)

  • Kim, ChangHwan;Kim, HyunWook;Kang, GilMo;Kim, GiYoung;Kim, WonHyuck;Park, ChanHong;Do, JongDae;Lee, MyoungHoon;Choi, SoonYoung;Park, HyeonYeong
    • Economic and Environmental Geology
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    • v.49 no.4
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    • pp.281-290
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    • 2016
  • Coastal areas, used as human utilization areas like leisure space, medical care, ports and power plants, etc., are regions that are continuously changing and interconnected with oceans and land. Regular monitoring of coastal changes is essential at key locations with such volatility. But the survey method of terrestial LiDAR(Light Detection and Ranging) system has much time consuming and many restrictions. For effective monitoring coastal changes, KIOST(Korea Institute of Ocean Science & Technology) has constructed a shipborne mobile LiDAR system. The shipborne mobile LiDAR system, installed in a research vessel, comprised a land based LiDAR(RIEGL LMS-420i), an IMU(MAGUS Inertial+), a RTKGNSS(LEICA GS15 GS25), and a fixed platform. The shipborne mobile LiDAR system is much more effective than a land based LiDAR system in the measuring of fore shore areas without shadow zone. Because the vessel with the shipborne mobile LiDAR system is continuously moved along the shoreline, it is possible to efficiently survey a large area in a relatively short time. We conducted test measurements in the Anmok-Songjung beach around the Gangneung port. Effective monitoring of the changes using the constructed shipborne mobile LiDAR system for seriously eroded coastal areas will be able to contribute to coastal erosion management and response.

Preventive Effect of Milk Products against Diabetes Mellitus Type 2: A Review (유제품의 제2형 당뇨병 예방 효과: 총설)

  • Kim, Dong-Hyeon;Chon, Jung-Whan;Kim, Hyun-Sook;Kim, Hong-Seok;Song, Kwang-Young;Kim, Young-Ji;Kang, Il-Byung;Lee, Soo-Kyung;Jeong, Dong-Gwan;Kim, Soo-Ki;Seo, Kun-Ho
    • Journal of Dairy Science and Biotechnology
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    • v.33 no.2
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    • pp.129-137
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    • 2015
  • Diabetes mellitus type 2 is a metabolic disorder that is characterized by hyperglycemia (high blood sugar level) in the context of insulin resistance and relative lack of insulin. Recently, much scientific evidence has shown that the risk of diabetes mellitus type 2 could be reduced by dairy intake. A significantly strong relationship has been noted between this disease and dairy intake. In particular, from the different types of fat in dairy foods that were reported to have a beneficial impact, low-fat dairy foods have been found to have the best effect with respect to reducing the risk of diabetes mellitus type 2. Therefore, the role of specific components of dairy foods, such as calcium, vitamin D, dairy fat, and trans-palmitoleic acid, which could be responsible for this effect and for the positive effect of dairy foods in obesity and metabolic syndrome, needs to be identified. There is a strong and relatively consistent body of accumulating evidence indicating that dairy foods may significantly reduce the risk of diabetes mellitus type 2, likely in a dose-response manner. Dairy recommendations should be an essential part of public health guidance, and identifying strategies to increase dairy food consumption to optimal levels is of utmost importance. Hence, this review summarizes various positive effects of dairy foods with respect to reducing the risk of diabetes mellitus type 2, based on available evidence, and discusses the need for further research on preventing or decreasing the risk of diabetes mellitus type 2.

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Phosphorus Removal Characteristics by Bacteria Isolated from Industrial Wastewater (산업폐수로부터 분리한 인제거 미생물의 인 제거 특성)

  • Kim, Hee-Jung;Lee, Seok-Eon;Hong, Hyeon-Ki;Kim, Deok-Hyun;An, Jung-Woo;Choi, Jong-Soon;Nam, Ju-Hyun;Lee, Moon-Soon;Woo, Sun-Hee;Chung, Keun-Yook
    • Korean Journal of Environmental Agriculture
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    • v.31 no.2
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    • pp.185-191
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    • 2012
  • Background: The removal of phosphate(P) in the wastewater is essential for the prevention of eutrophication in the river and stream. This study was initiated to evaluate the P removal by three strains of bacteria isolated from industrial wastewater. The three strains of bacteria, A1, A2, and A3, isolated were identified as Stenotrophomonas maltophilia strain CUPS 3, Rhodococcus erythropolis strain Sco-C01, Bacillus sp. 3434BRRJ, respectively. METHODS AND RESULTS: The experiments evaluating the effects of temperature, P concentration, aeration, and carbon sources on P removal by Bacillus sp. 3434BRRJ were performed in the following conditions: temperature, 15, 25 and $30^{\circ}C$; P concentrations, 20, 30, and 40 mg/L; oxygen condition, aerobic, anaerobic/aerobic conditions; carbon sources, glucose, acetate and mixture of glucose and acetate. As a result, the best optimum conditions for P removal by Bacillus sp. 3434BRRJ were as follows: temperature, $30^{\circ}C$; P concentration, 20 mg/L; carbon sources, mixture of glucose and acetate; oxygen concentration, anaerobic and aerobic conditions. The P removal efficiencies by Bacillus sp. 3434BRRJ, Stenotrophomonas maltophilia strain CUPS, and Rhodococcus erythropolis strain Sco-C01 were 99%, 50%, 20%, respectively. CONCLUSION: As a result, the best optimum conditions for P removal by Bacillus sp. 3434BRRJ selected and used in this study were as follows: temperature, $30^{\circ}C$; P concentration, 20 mg/L; carbon sources, mixture of glucose and acetate; oxygen concentration, anaerobic and aerobic conditions.

Control of Mg and P Ion Concentration as a Precondition to Use N, K and Ca Ion Sensors in Closed Hydroponics (N, K, Ca의 한정된 이온센서 이용을 전제로 한 순환식 수경재배에서 P, Mg의 조절 방법)

  • Choi, Gyeong Lee;Yeo, Kyung Hwan;Rhee, Han Cheol;Lee, Seong Chan;Lee, Jung-Sup;Kang, Nam Jun;Kim, Hak Jin;Jung, Dae Hyun
    • Horticultural Science & Technology
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    • v.34 no.6
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    • pp.871-877
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    • 2016
  • Recycling nutrient solutions in closed hydroponic production systems is usually accompanied by an imbalance of nutrient solutions when concentration is controlled according to electrical conductivity (EC) levels. This study investigated whether it was possible to automatically control the concentrations of five essential elements nitrogen (N), phosphorus (P), potassium (K), calcium (Ca) and magnesium (Mg) using only N, K and Ca ion sensors. N, P, K, Ca, and Mg uptake was measured in the nutrient solution, and relationships between absorbed ions were analyzed through twice-repeated experiments in lettuce. Results confirmed that the pattern of $PO_4$ ion uptake was similar that of N, and the pattern of Mg ion uptake was similar that of Ca. $PO_4$ ion uptake was most highly correlated with N, and Mg was most highly correlated with Ca. Regression coefficients of N and $PO_4$ were significantly different at 1.04 and 0.55, respectively, but were similar between Ca and Mg at 0.35 and 0.40, respectively. Additional experiments were conducted to measure nutrient uptake in pak choi and rose plants, both to confirm the results from the first experiment in lettuce, and to assess possible application to other crops. Coefficients of determination both for N and $PO_4$, and Ca and Mg were considerably high ($R^2=0.86$) in cultured pak choi, and similar results were observed in cultured rose ($R^2=0.87$ and 0.73, respectively). Regression coefficients for cultured pak choi were 0.56 and 0.24, respectively, and for rose were 0.51 and 0.16, respectively. Although the results obtained for N and $PO_4$ were not consistent between the lettuce experiments, N and $PO_4$ have similar regression coefficients for all crops. No common coefficient was found between Ca and Mg.