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Efficient (3R)-Acetoin Production from meso-2,3-Butanediol Using a New Whole-Cell Biocatalyst with Co-Expression of meso-2,3-Butanediol Dehydrogenase, NADH Oxidase, and Vitreoscilla Hemoglobin

  • Guo, Zewang;Zhao, Xihua;He, Yuanzhi;Yang, Tianxing;Gao, Huifang;Li, Ganxin;Chen, Feixue;Sun, Meijing;Lee, Jung-Kul;Zhang, Liaoyuan
    • Journal of Microbiology and Biotechnology
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    • v.27 no.1
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    • pp.92-100
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    • 2017
  • Acetoin (AC) is a volatile platform compound with various potential industrial applications. AC contains two stereoisomeric forms: (3S)-AC and (3R)-AC. Optically pure AC is an important potential intermediate and widely used as a precursor to synthesize novel optically active materials. In this study, chiral (3R)-AC production from meso-2,3-butanediol (meso-2,3-BD) was obtained using recombinant Escherichia coli cells co-expressing meso-2,3-butanediol dehydrogenase (meso-2,3-BDH), NADH oxidase (NOX), and hemoglobin protein (VHB) from Serratia sp. T241, Lactobacillus brevis, and Vitreoscilla, respectively. The new biocatalyst of E. coli/pET-mbdh-nox-vgb was developed and the bioconversion conditions were optimized. Under the optimal conditions, 86.74 g/l of (3R)-AC with the productivity of 3.61 g/l/h and the stereoisomeric purity of 97.89% was achieved from 93.73 g/l meso-2,3-BD using the whole-cell biocatalyst. The yield and productivity were new records for (3R)-AC production. The results exhibit the industrial potential for (3R)-AC production via whole-cell biocatalysis.

Analysis of Spatial Variability in a Korean Paddy Field Using Median Polish Detrending (Median polish 기법을 이용한 한국 논의 공간변이 분석)

  • Chung, Sun-Ok;Jung, In-Kyu;Sung, Je-Hoon;Sudduth, Kenneth A.;Drummond, Scott T.
    • Journal of Biosystems Engineering
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    • v.33 no.5
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    • pp.362-369
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    • 2008
  • There is developing interest in precision agriculture in Korea, despite the fact that typical Korean fields are less than 1 ha in size. Describing within-field variability in typical Korean production settings is a fundamental first step toward determining the size of management zones and the inter-relationships between limiting factors, for establishment of site-specific management strategies. Measurements of rice (Oriza Sativa L) yield, chlorophyll content, and soil properties were obtained in a small (100-m by 30-m) Korean rice paddy field. Yield data were manually collected on 10-m by 5-m grids (180 samples with 3 samples in each of 60 grid cells) and chlorophyll content was measured using a Minolta SPAD 502 in 2-m by 2-m grids. Soil samples were collected at 275 points to compare results from sampling at different scales. Ten soil properties important for rice production in Korea were determined through laboratory analyses. Variogram analysis and point kriging with and without median polishing were conducted to determine the variability of the measured parameters. Influence of variogram model selection and other parameters on the interpretation of the data was investigated. For many of the data, maximum values were greater than double the minimum values, indicating considerable spatial variability in the small paddy field, and large-scale spatial trends were present. When variograms were fit to the original data, the limits of spatial dependency for rice yield and SP AD reading were 11.5 m and 6.5 m, respectively, and after detrending the limits were reduced to 7.4 m and 3.9 m. The range of spatial dependency for soil properties was variable, with several having ranges as short as 2 m and others having ranges greater than 30 m. Kriged maps of the variables clearly showed the presence of both large-scale (trend) variability and small-scale variability in this small field where it would be reasonable to expect uniformity. These findings indicate the potential for applying the principles and technology of precision agriculture for Korean paddy fields. Additional research is needed to confirm the results with data from other fields and crops.d similar tendency with the result for the frequency less than 20 Hz, but the width of change was reduced highly.

Improving Microalgal Biomass Productivity and Preventing Biofouling in Floating Marine Photobioreactors via Sulfonation of Selectively Permeable Membranes (부유형 해양 광생물반응기의 선택적 투과막의 술폰화 반응을 통한 Biofouling 억제 및 미세조류 생산성 향상)

  • Kim, Kwangmin;Lee, Yunwoo;Kim, Z-Hun;Park, Hanwool;Jung, Injae;Park, Jaehoon;Lim, Sang-Min;Lee, Choul-Gyun
    • Journal of Marine Bioscience and Biotechnology
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    • v.9 no.1
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    • pp.14-21
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    • 2017
  • The purpose of this study was to inhibit biofouling on a selectively permeable membrane (SPM) and increase biomass productivity in marine photobioreactors (PBRs) for microalgal cultivation by chemical treatment. Surfaces of a SPM, composed of polyethylene terephthalate (PET), was sulfonated to decrease hydrophobicity through attaching negatively charged sulfonic groups. Reaction time of sulfonation was varied from 0 min to 60 min. As the reaction time increased, the water contact angle value of SPM surface was decreased from $75.5^{\circ}$ to $44.5^{\circ}$, indicating decrease of surface hydrophobicity. Furthermore, the water permeability of sulfonated SPM was increased from $5.42mL/m^2/s$ to $10.58mL/m^2/s$, which reflects higher nutrients transfer rates through the membranes, due to decreased hydrophobicity. When cultivating Tetraselmis sp. using 100-mL floating PBRs with sulfonated SPMs, biomass productivity was improved by 34% compared with the control group (non-reacted SPMs). In addition, scanning electron microscopic observation of SPMs used for cultivation clearly revealed lower degree of cell attachment on the sulfonated SPMs. These results suggest that sulfornation of a PET SPM could improve microalgal biomass productivity by increasing nutrients transfer rates and inhibiting biofouling by algal cells.

Effects of Endomycorrhizal Glomus Inoculation on Drought Resistance and Physiological Changes of Lespedeza cyrtobotrya Seedlings Exposed to Water Stress (Glomus 내생균근균(內生菌根菌) 접종(接種)이 수분(水分) 스트레스에 노출(露出)된 참싸리의 건조저항성(乾燥抵抗性) 및 생리적(生理的) 변화(變化)에 미치는 영향(影響))

  • Kim, Hyo-Jin;Lee, Kyung-Joon;Han, Sim-Hee
    • Journal of the Korean Society of Environmental Restoration Technology
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    • v.5 no.2
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    • pp.53-62
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    • 2002
  • The objectives of this study were to understand the tolerance mechanism of woody plants to water stress and tolerance changes in relation to mycorrhizal formation. Lespedeza cyrtobotrya Miq. commonly used for erosion control in slopes were raised from seeds and transplanted to 120 plastic pots. Sixty pots received the top soil of a Fraxinus americana forest, while remaining 60 pots received the autoclaved top soil. The forest soil contained 1,200 spores per 100g of arbuscular endomycorrhizal fungus, mostly Glomus sp. The plants were raised outside with regular supply of water and mineral nutrients. Two kinds of water deficit treatment and a control were started at the middle of July : cyclic water deficit treatment with 3 cycles of sequential water stress at the point of xylem water potential of about -0.6, -0.6, and -1.7 MPa and recovery, and non-cyclic water deficit treatment with single water stress at about -1.5 MPa. The non-stressed plants received plenty of water throughout the period. In late August the plants were harvested for measurements of dry weight, N, P, carbohydrate contents, net photosynthesis and superoxide dismutase(SOD) activities. Both cyclic and non-cyclic water deficit treatments reduced dry weight by 60% and 40%, respectively, and reduced nitrogen absorption, while increased SOD activities. Water-stressed plants also showed increased carbohydrate contents in the leaves and lowered stomatal conductance. Mycorrhizal inoculation resulted in an average of 40% infection of roots and 2-3 times increase in P absorption in water-stressed as well as non-stressed plants. Mycorrhizal formation also increased shoot-root ratio. The results that SOD activities of water-stressed plants with mycorrhizal infection were significantly lower than those of non-mycorrhizal plants suggest the possibility of improvement of water-stressed condition by mycorrhizal formation. It was concluded that endomycorrhizal formation increased tolerance of Lespedeza cyrtobotrya seedlings to water stress.

Quantitative Analysis of Bacillus amyloliquefaciens GR4-5 in Soil (Bacillus amyloliquefaciens GR4-5 균주의 토양 내 정량 분석)

  • Kim, Dayeon;Kim, Byung-Yong;Ahn, Jae-Hyung;Weon, Hang-Yeon;Kim, Sung-Il;Kim, Wan-Gyu;Song, Jaekyeong
    • Korean Journal of Organic Agriculture
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    • v.23 no.4
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    • pp.847-858
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    • 2015
  • Bacillus amyloliquefaciens GR4-5 was isolated from the rhizosphere soil of Korean ginseng and displayed broad-spectrum suppression of ginseng root rot pathogens. The survivability of B. amyloliquefaciens GR4-5 in soil was investigated under three different conditions; indoor, outdoor - of which soil was put in 14 mL tube after treatment - and field environments. Soil samples were collected over a four-week period from three experimental designs, and assessed for 16S rRNA gene copy number by quantitative polymerase chain reaction (qPCR). In outdoor condition, the 16S rRNA gene copy number of Bacillus spp. was 8.35 log copies g $soil^{-1}$ immediately after the GR4-5 treatment. Two weeks later, the 16S rRNA gene copy number of Bacillus spp. (6.70 log copies g $soil^{-1}$) was similar to that of the control (6.38 log copies g $soil^{-1}$). In indoor condition, the 16S rRNA gene copy number of Bacillus spp. maintained in a certain level for a longer period than those in outdoor and field. The 16S rRNA gene copy number of Bacillus spp. in field experiment was reduced faster than that of outdoor condition. Our results show that B. amyloliquefaciens GR4-5 can survive in bulk soil for 1 week, indicating its potential use as a biocontrol agent following 7 day application intervals. This study presents that outdoor microcosm system design could be a useful method to assess easily the survivability of beneficial microorganisms.

Expression of Ang-2/Tie-2 and PI3K/AKT in Colorectal Cancer

  • Zhang, Ji-Hong;Wang, Li-Hua;Li, Xiang-Jun;Wang, Ai-Ping;Reng, Li-Qun;Xia, Feng-Guo;Yang, Zhi-Ping;Jiang, Jing;Wang, Xiao-Dan;Wen, Chun-Yang
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.20
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    • pp.8651-8656
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    • 2014
  • Purpose: To study the expression of angiogenin-2 (Ang-2) and its receptor Tie-2 in colorectal cancer and discuss the possible mechanisms behind this process. Materials and Methods: Using the streptavidin-peroxidase (SP) immunohistochemical method, paraffin sections from 100 colorectal cancer samples and 10 samples from tumor-adjacent normal tissue (> 2 cm from the edge of the gross tumor) were tested for protein expression of Ang-2, Tie-2, PI3K, and AKT. Reverse transcription-polymerase chain reaction and Western blots were further used to measure expression of the 4 genes and proteins in 20 freshly-resected colorectal cancer samples and tumor-adjacent normal tissues. Results: In colorectal cancer tissues, the expression of the Ang-2, Tie-2, PI3K, and AKT genes and their proteins was significantly higher than in tumor-adjacent normal tissues. Protein expression in poorly-differentiated adenocarcinoma was higher than that in well and moderately differentiated adenocarcinoma. According to Duke's classification, the protein expression in Stages C and D was significantly higher than that in Stages A and B. In the group with lymphatic metastasis, the protein expression was higher than that without lymphatic metastasis. Conclusions: In colorectal cancer, the expression of the Ang-2, Tie-2, PI3K, and AKT genes and their proteins is markedly higher than those in tumor-adjacent normal tissues. No correlation was observed between protein expression and gender, location, or histologic type. Correlations did exist between protein expression and differentiation level, stage of Duke's classification, and lymphatic metastasis; in colorectal cancer tissues with lower differentiation levels, higher stages of Duke's classification, and lymphatic metastasis, the expression of all 4 proteins was higher. The study of their expression patterns and relationships with aggression and metastasis will provide a valuable experimental foundation for assessing prognosis and targeted therapy of colorectal cancer.

Effects of IFN-γ on IL-18 Expression in Pregnant Rats and Pregnancy Outcomes

  • Si, Li-Fang;Zhang, Shou-Yan;Gao, Chun-Sheng;Chen, Shu-Lin;Zhao, Jin;Cheng, Xiang-Chao
    • Asian-Australasian Journal of Animal Sciences
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    • v.26 no.10
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    • pp.1399-1405
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    • 2013
  • The present study focused on establishing the effects of interferon-gamma (IFN-${\gamma}$) on interleukin-18 (IL-18) expression patterns and pregnancy outcomes in pregnant rats. Pregnant rats at the post-implantation stage were randomized into control, low IFN-${\gamma}$ (L-IFN-${\gamma}$) and high IFN-${\gamma}$ groups (H-IFN-${\gamma}$) that received normal saline, 100 IU/g of IFN-${\gamma}$ and 500 IU/g of IFN-${\gamma}$ vaginal muscular injection, respectively. The effects of IFN-${\gamma}$ on IL-18 expression and pregnancy outcomes were assessed systematically using several methods, including immunohistochemistry streptavidin-perosidase (SP), image pattern analysis, enzyme-linked immune-sorbent assay (ELISA), whole blood count (WBC) count, microscopy and visual observation. IL-18 was detected in the uteri of all pregnant rats, and mainly distributed in the endometrium, decidual cells, vascular endothelium and myometrium. Immunohistochemistry and image pattern analyses revealed significantly lower IL-18 expression in the H-IFN-${\gamma}$ group compared to the L-IFN-${\gamma}$ and control groups (p<0.01), indicating that high doses of IFN-${\gamma}$ induce downregulation of IL-18 in the uterus of pregnant rats. ELISA results disclosed that IL-18 expression in peripheral blood of the H-IFN-${\gamma}$ group was lower than that of the L-IFN-${\gamma}$ group (p<0.05), and significantly reduced compared to the control group (p<0.01). Moreover, the number of peripheral leukocytes in the H-IFN-${\gamma}$ group was significantly higher than those in the control and L-IFN-${\gamma}$ groups (p<0.01). Morphology analysis showed no evident differences between the L-IFN-${\gamma}$ and control groups. However, for the H-IFN-${\gamma}$ group, uterine mucosa bleeding, necrosis and excoriation were observed using microscopy. Visual observation revealed marroon, swelling, crassitude and no embryo in the uterus, which are obvious indicators of abortion. These results indicate that IFN-${\gamma}$ plays a regulatory role in IL-18 expression in the uterus and peripheral blood of pregnant rats at the post-implantation stage. Moreover, high levels (500 IU/g) of IFN-${\gamma}$ influence normal pregnancy at the early stages in rats by downregulating IL-18 expression in the uterus and peripheral blood and increasing the number of peripheral leukocytes, consequently triggering termination of pregnancy.

Hexagonal Boron Nitride Monolayer Growth without Aminoborane Nanoparticles by Chemical Vapor Deposition

  • Han, Jaehyu;Yeo, Jong-Souk
    • Proceedings of the Korean Vacuum Society Conference
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    • 2014.02a
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    • pp.409-409
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    • 2014
  • Recently hexagonal boron nitride (h-BN), III-V compound of boron and nitrogen with strong covalent $sp^2$ bond, is a 2 dimensional insulating material with a large direct band gap up to 6 eV. Its outstanding properties such as strong mechanical strength, high thermal conductivity, and chemical stability have been reported to be similar or superior to graphene. Because of these excellent properties, h-BN can potentially be used for variety of applications such as dielectric layer, deep UV optoelectronic device, and protective transparent substrate. Ultra flat and charge impurity-free surface of h-BN is also an ideal substrate to maintain electrical properties of 2 dimensional materials such as graphene. To synthesize a single or a few layered h-BN, chemical vapor deposition method (CVD) has been widely used by using an ammonia borane as a precursor. Ammonia borane decomposes into hydrogen (gas), monomeric aminoborane (solid), and borazine (gas) that is used for growing h-BN layer. However, very active monomeric aminoborane forms polymeric aminoborane nanoparticles that are white non-crystalline BN nanoparticles of 50~100 nm in diameter. The presence of these BN nanoparticles following the synthesis has been hampering the implementation of h-BN to various applications. Therefore, it is quite important to grow a clean and high quality h-BN layer free of BN particles without having to introduce complicated process steps. We have demonstrated a synthesis of a high quality h-BN monolayer free of BN nanoparticles in wafer-scale size of $7{\times}7cm^2$ by using CVD method incorporating a simple filter system. The measured results have shown that the filter can effectively remove BN nanoparticles by restricting them from reaching to Cu substrate. Layer thickness of about 0.48 nm measured by AFM, a Raman shift of $1,371{\sim}1,372cm^{-1}$ measured by micro Raman spectroscopy along with optical band gap of 6.06 eV estimated from UV-Vis Spectrophotometer confirm the formation of monolayer h-BN. Quantitative XPS analysis for the ratio of boron and nitrogen and CS-corrected HRTEM image of atomic resolution hexagonal lattices indicate a high quality stoichiometric h-BN. The method presented here provides a promising technique for the synthesis of high quality monolayer h-BN free of BN nanoparticles.

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The Change of c-jun Promoter Activity in TPA-Induced U937 Cells Infected with Human Cytomegalovirus (HCMV) (TPA로 분화된 U937 세포에서 사람 세포거대바이러스에 의한 c-jun Promoter 활성도의 변화)

  • Park, Chung-Gyu;Kim, Dae-Joong;Kim, Jin-Hee;Han, Tae-Hee;Hwan, Eung-Soo;Choi, Myong-Sik;Kook, Yoon-Hoh;Choi, Sung-Bae;Cha, Chang-Yong
    • The Journal of Korean Society of Virology
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    • v.29 no.2
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    • pp.129-136
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    • 1999
  • Transient transfection assay has been done to evaluate whether the c-jun activation would be prerequisite to the induction of permissiveness against human cytomegalovirus using in vitro cell model in which U937 has been induced to express CD11b and CD14 to become potential monocyte/macrophage cells by TPA treatment. U937 cells were treated with $10\;{\mu}M$, $50\;{\mu}M$ or $100\;{\mu}M$ of TPA. The cell morphology change was observed and the expression of the CD11b and CD14 was confirmed by FACS. Differentiated cells were transfected with pJLuc reporter vector which contained the wild type murine c-jun promoter spanning the SP1, CTF, ATF/CREB and MEF-2 binding sites upstream of the firefly luciferase gene. After 48 hrs of transfection, the cells were infected with HCMV Towne strain and the luciferase activity was assessed at 1 hand 4 h pi. The transfection assay showed no activation of the c-jun promoter at 1 h pi, instead, it showed 2 times increase of the its activity at 4 h pi. There was no difference of the c-jun promoter activation between TPA treated and untreated U937 cells, implying that c-jun activation might not be prerequisite for allowing cells to be premissive to HCMV, although HCMV infection itself could activate c-jun promoter.

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Analysis of Trans Fatty Acid Content in Retort Food, Powdered Milk, Biscuit and Pizza Products (레토르트식품, 분유, 비스킷 및 피자 내에 함유되어 있는 트랜스지방산 함량 분석)

  • Park, Da-Jung;Park, Jung-Min;Shin, Jin-Ho;Song, Jae-Cheol;Kim, Jin-Man
    • Food Science of Animal Resources
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    • v.28 no.2
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    • pp.240-245
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    • 2008
  • The consumption of foods containing trans fatty acids (TFAs) is a matter of concern at present. According to many studies, trans fatty acids (TFAs) may cause illnesses such as the coronary heart disease, diabetes mellitus, large intestine cancer, and breast cancer. They can also raise low density lipoprotein (LDL) cholesterol and reduce high density lipoprotein (HDL) cholesterol. TFAs can also inhibit the synthesis of phospholipids containing polyunsaturated fatty acids in arterial cells. As a consequence the Food and Drug Administration has deemed that saturated fatty acid, cholesterol and trans fatty acid levels be listed on food labels as of 2006. The Korea Food and Drug Administration also has required the listing of trans fatty acid content on food labels since 2007. The aim of this study was to determine the total lipid and trans fatty acid (TFA) contents in retort food, powdered milk, biscuit and pizza products. The number of samples examined were 2 retort food, 6 powdered milk, 7 biscuit and 3 pizza products. The extraction of total lipids in retort food and powdered milk followed the chloroform methanol method. The extraction of total lipids in biscuit and pizza was by the acid digestion method. All samples were analyzed by gas chromatography (GC) using a SP-2560 capillary column and a flame ionization detector. The TFA contents per 100g of sample were 1-2.8% (1.9%) in retort foods, 0.4-2.4% (1.37%) in powdered milk products, 0-2.9% (1.23%) in biscuits, and 2.8-3.45% (3.03%) in pizzas.