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Modification of N-Terminal Amino Acids of Fungal Benzoate Hydroxylase (CYP53A15) for the Production of p-Hydroxybenzoate and Optimization of Bioproduction Conditions in Escherichia coli

  • Tamaki, Shun;Yagi, Mitsuhiko;Nishihata, Yuki;Yamaji, Hideki;Shigeri, Yasushi;Uno, Tomohide;Imaishi, Hiromasa
    • Journal of Microbiology and Biotechnology
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    • v.28 no.3
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    • pp.439-447
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    • 2018
  • The aromatic compound p-hydroxybenzoate (PHBA) is an important material with multiple applications, including as a building block of liquid crystal polymers in chemical industries. The cytochrome P450 (CYP) enzymes are beneficial monooxygenases for the synthesis of chemicals, and CYP53A15 from fungus Cochliobolus lunatus is capable of executing the hydroxylation from benzoate to PHBA. Here, we constructed a system for the bioconversion of benzoate to PHBA in Escherichia coli cells coexpressing CYP53A15 and human NADPH-P450 oxidoreductase (CPR) genes as a redox partner. For suitable coexpression of CYP53A15 and CPR, we originally constructed five plasmids in which we replaced the N-terminal transmembrane region of CYP53A15 with a portion of the N-terminus of various mammalian P450s. PHBA productivity was the greatest when CYP53A15 expression was induced at $20^{\circ}C$ in $2{\times}YT$ medium in host E. coli strain ${\Delta}gcvR$ transformed with an N-terminal transmembrane region of rabbit CYP2C3. By optimizing each reaction condition (reaction temperature, substrate concentration, reaction time, and E. coli cell concentration), we achieved 90% whole-cell conversion of benzoate. Our data demonstrate that the described novel E. coli bioconversion system is a more efficient tool for PHBA production from benzoate than the previously described yeast system.

Effect of spinning parameters of polyethersulfone based hollow fiber membranes on morphological and mechanical properties

  • Tewfik, Shadia R.;Sorour, Mohamed H.;Shaalan, Hayam F.;Hani, Heba A.
    • Membrane and Water Treatment
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    • v.9 no.1
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    • pp.43-51
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    • 2018
  • Hollow fiber (HF) membranes are gaining wide interest over flat membranes due to their compaction and high area to surface volume ratio. This work addresses the fabrication of HF from polysulfone (PS) and polyethersulfone (PES) using N-methylpyrrolidone (NMP) as solvent in addition to other additives to achieve desired characteristics. The semi-pilot spinning system includes jacketed vessel, four spinneret block, coagulation and washing baths in addition to dryer and winder. Different parameters affecting dry-wet spinning phase inversion process were investigated. Dope compositions of PES, NMP and polyvinyl pyrrolidone (PVP) of varying molecular weights as additive were addressed. Some critical parameters of importance were also investigated. Those include dope flow rate, air gap, coagulation & washing baths and drying temperatures. The measured dope viscosity was in the range from 1.7 to 36.5 Pa.s. Air gap distance was adjusted from 20 to 45 cm and coagulation bath temperature from 20 to $46^{\circ}C$. The HF membranes were characterized by scanning electron microscope (SEM), atomic force microscope (AFM) and mechanical properties. Results indicated prevalence of finger like structure and average surface roughness from about 29 to 78.3 nm. Profile of stress strain characteristics revealed suitability of the fibers for downstream interventions for fabrication of thin film composite membrane. Different empirical correlations were formulated which enable deeper understanding of the interaction of the above mentioned variables. Data of pure water permeability (PWP) confirmed that the fabricated samples fall within the microfiltration (MF)-ultrafiltration (UF) range of membrane separation.

Development of Precision Drilling Machine for the Instrumentation of Nuclear Fuels (핵연료계장을 위한 정밀 드릴링장치 개발)

  • Hong, Jintae;Jeong, Hwang-Young;Ahn, Sung-Ho;Joung, Chang-Young
    • Journal of the Korean Society for Precision Engineering
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    • v.30 no.2
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    • pp.223-230
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    • 2013
  • When a new nuclear fuel is developed, an irradiation test needs to be carried out in the research reactor to analyze the performance of the new nuclear fuel. In order to check the performance of a nuclear fuel during the irradiation test in the test loop of a research reactor, sensors need to be attached in and out of the fuel rod and connect them with instrumentation cables to the measuring device located outside of the reactor pool. In particular, to check the temporary temperature change at the center of a nuclear fuel during the irradiation test, a thermocouple should be instrumented at the center of the fuel rod. Therefore, a hole needs to be made at the center of fuel pellet to put in the thermocouple. However, because the hardness and the density of a sintered $UO_2$ pellet are very high, it is difficult to make a small fine hole on a sintered $UO_2$ pellet using a simple drilling machine even though we use a diamond drill bit made by electro deposition. In this study, an automated drilling machine using a CVD diamond drill has been developed to make a fine hole in a fuel pellet without changing tools or breakage of workpiece. A sintered alumina ($Al_2O_3$) block which has a higher hardness than a sintered $UO_2$ pellet is used as a test specimen. Then, it is verified that a precise hole can be drilled off without breakage of the drill bit in a short time.

Effects of Biofertilizer on the Quality and Antioxidant Property of Rice (Oryza sativa L.)

  • Rico, Cyren Mendoza;Bhuiyan, Mohammad Kamrul Islam;Mintah, Lemuel O.;Shin, Dong-Il;Chung, Il-Kyung;Son, Tae-Kwon;Lee, Sang-Chul
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.52 no.3
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    • pp.274-280
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    • 2007
  • The effect of biofertilizer in enhancing nutrient quality and antioxidant property of rice grain was investigated. The experiment was carried out in a randomized complete block design with 3 replications and 7 treatments namely : RF = $N-P_2O_5-K_2O(11-5.5-4.8kg\;10a^{-1});$ half of the recommended fertilizer rate, $HRF=N-P_2O_5-K_2O(5.5-2.75-2.4kg\;10a^{-1}):$ HRF+Bio 250=HRF combined with 250 kg Biofertilizer 10 $a^{-1}$; HRF+Bio 500=HRF combined with 500 kg Biofertilizer 10 $a^{-1};$ Bio 250=250 kg Biofertilizer 10 $a^{-1};$ Bio 500=500 kg Biofertilizer 10 $a^{-1};$ and NF=No Fertilizer. Results showed that HRF+Bio 500 obtained a significantly higher protein content but a significantly lower amylose content compared with RF and NF treatments. Highest phytic acid content was recorded in NF treatment while the lowest was observed in HRF+500 treatment. The highest values in both electron donating ability and reducing power were obtained in HRF+Bio 500 treatment. All treatments obtained higher reducing power than that of the RF treatment and that NF treatment showed comparable values in both electron donating ability and reducing power with those of the treated plots. Highest antimutagenicity property was also observed in HRF+Bio 500 treatment followed by Bio 500 treatment. This study showed the possibility of using biofertilizer to enhance nutritional quality and antioxidant property of rice.

Effects of Mixed Application of Wood Vinegar and Herbicides on Weed Control, Yield and Quality of Rice(Oryza sativa L.)

  • Rico, Cyren M.;Souvandouane, Souliya;Mintah, Lemuel Ohemeng;Chung, Il-Kyung;Son, Tae-Kwon;Lee, Sang-Chul
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.52 no.4
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    • pp.387-392
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    • 2007
  • The effect of mixed treatments of wood vinegar and sulfonylurea-based herbicides on weed control, yield and yield components, and quality of rice was investigated. Two herbicides were tested namely: imazosulfuron-ethyl+thiobencarb[ethyl-1-(2-chloroimidazo[1,2-$\alpha$]pyridin-3-ylsulfonyl)-3-(4,6-dimethoxypyrimidin-2-yl) urea+S-4-chlorobenzyl diethyl(thiocarbamate)], and bensulfuronmethyl+butachlor [methyl $\alpha$-[(4,6-dimethoxypyrimidin-2-ylcarbamoyl)sulfamoyl]-o-toluate+N-butoxymethyl-2-chloro-2',6'-diethylacetanilide]. The experiment was carried out in a randomized complete block design with 3 replications and 5 treatments. Treatments used were recommended(RH: 100%) and half-recommended(HRH: 50%) application rates of each herbicide. Half-recommended application rates were combined with 1 mL wood vinegar $500mL\;water^{-1}$(500) and 1 mL wood vinegar $1000mL\;water^{-1}$(1000) wood vinegar. Plots for no herbicide treatments were also prepared and used as control. Results showed that wood vinegar significantly increased efficacy of HRH in bensulfuron-methyl+butachlor while high efficacy was already obtained in HRH treatment of imazosulfuron-ethyl+thiobencarb. Wood vinegar did not improve the efficacy of imazosulfuron-ethyl+thiobencarb but improved rice yield. Significantly similar rice yields were obtained in the HRH+1000 WV and RH treatments of both herbicides. There were no significant variations in the yield components among the treatments; however, differences in yield can be attributed to the variations in the spikelet number and ripening ratio. Data on rice quality analysis did not show clear trend on the effects of the treatments on grain appearance and nutritional quality.

Predictors of Intensive Care Unit Morbidity and Midterm Follow-up after Primary Repair of Tetralogy of Fallot

  • Egbe, Alexander C.;Nguyen, Khanh;Mittnacht, Alexander J.C.;Joashi, Umesh
    • Journal of Chest Surgery
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    • v.47 no.3
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    • pp.211-219
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    • 2014
  • Background: Our objectives were to review our institutional early and midterm experience with primary tetralogy of Fallot (TOF) repair, and identify predictors of intensive care unit (ICU) morbidity. Methods: We analyzed perioperative and midterm follow-up data for all cases of primary TOF repair from 2001 to 2012. The primary endpoint was early mortality and morbidity, and the secondary endpoint was survival and functional status at follow-up. Results: Ninety-seven patients underwent primary repair. The median age was 4.9 months (range, 1 to 9 months), and the median weight was 5.3 kg (range, 3.1 to 9.8 kg). There was no early surgical mortality. The incidence of junctional ectopic tachycardia and persistent complete heart block was 2% and 1%, respectively. The median length of ICU stay was 6 days (range, 2 to 21 days), and the median duration of mechanical ventilation was 19 hours (range, 0 to 136 hours). By multiple regression analysis, age and weight were independent predictors of the length of ICU stay, while the surgical era was an independent predictor of the duration of mechanical ventilation. At the 8-year follow-up, freedom from death and re-intervention was 97% and 90%, respectively. Conclusion: Primary TOF repair is a safe procedure with low mortality and morbidity in a medium-sized program with outcomes comparable to national standards. Age and weight at the time of surgery remain significant predictors of morbidity.

Synthesis and Evaluation of a Ligand Targeting the Somatostatin Receptor for Drug Delivery to Tumor Cell (암세포 내로의 약물 전달 증진 목적의 신규 소마토스타틴 수용체 타겟리간드 합성 및 평가)

  • Choi, SunJu;Hong, YoungDon;Lee, SoYoung;Jung, SungHee
    • Journal of Radiation Industry
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    • v.9 no.4
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    • pp.193-198
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    • 2015
  • Most of targeted therapies block the action of certain enzymes, proteins, or other molecules involved in the growth and spread of cancer cells to produce its cytotoxic effect. Either small molecule drugs or monoclonal antibodies are mostly used in targeted therapies. Unfortunately, targeted therapy has a certain degree of unwanted side effect like other cytotoxicity inducing chemotherapies. To overcome and to reduce unwanted side effects during a cancer therapy, recently radiopeptide therapies has got the worlds' attraction for the tumor targeting modalities due to its beneficial effect on less side effect compared to cytotoxic chemotherapies. Among radiopeptide therapies, $^{177}Lu$-DOTATATE is a major modality as an effective one invented so far in treating neuroendocrine tumor (NET) and it has been in clinical trials at least one decade. Although it does have rather effective therapeutic effect on NET, it has less effective in rather large solid tumor. There are many ways to improve or increase therapeutic effect of radiopeptide are a finding the potent small molecules to target the tumor site selectively, or a labeling with radioisotope of emitting high energy, or an improving its biological half-life by introducing different moieties to increase lipophilicity. Present study was focus to increase a biological half-life of radio somatostatin which will target the somatostatin receptor by altering the bifunctional chelator (BFCA) by introducing lipophilic moiety to the somatostatin, which would make the labeled peptide stay longer in the tumor site and thus it can intensify the therapeutic effect on tumor cell itself and around tissues.

Long-Term Monitoring of the Barrier Effect of the Wild Boar Fence

  • Lim, Sang Jin;Kwon, Ji Hyun;Namgung, Hun;Park, Joong Yeol;Kim, Eui Kyeong;Park, Yung Chul
    • Journal of Forest and Environmental Science
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    • v.38 no.2
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    • pp.128-132
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    • 2022
  • Wild boars (Sus scrofa) not only cause crop damage and human casualties, but also facilitate the spread of many infectious diseases in domestic animals and humans. To determine the efficiency of a fencing system in blocking the movement of wild boars, long-term monitoring was performed in a fenced area in Bukhansan National Park using camera traps. Upon monitoring for a period of 46 months, there was a 72.6% reduction in the number of wild boar appearances in the fence-enclosed area, compared to that in the unenclosed area. For 20 months after the fence installation, the blocking effect of the fence was effective enough to reduce the appearance of wild boars by 92.6% in the fence-enclosed area, compared to that in the unenclosed area. The blocking effect of the fence remained effective for 20 months after its installation, after which its effectiveness decreased. Maintaining a fence for a long time is likely to lead to habitat fragmentation. It can also block the movement of other wild animals, including the endangered species - the long-tailed goral. This study suggests a 20-month retention period for the fences installed to inhibit the movement of wild boars in wide forests such as Gangwon-do in South Korea. To identify how long the blocking effect of the fences lasts, further studies are needed focusing on the length and height of the fence, and the conditions of the ground surface.

Design and Architecture of Low-Latency High-Speed Turbo Decoders

  • Jung, Ji-Won;Lee, In-Ki;Choi, Duk-Gun;Jeong, Jin-Hee;Kim, Ki-Man;Choi, Eun-A;Oh, Deock-Gil
    • ETRI Journal
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    • v.27 no.5
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    • pp.525-532
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    • 2005
  • In this paper, we propose and present implementation results of a high-speed turbo decoding algorithm. The latency caused by (de)interleaving and iterative decoding in a conventional maximum a posteriori turbo decoder can be dramatically reduced with the proposed design. The source of the latency reduction is from the combination of the radix-4, center to top, parallel decoding, and early-stop algorithms. This reduced latency enables the use of the turbo decoder as a forward error correction scheme in real-time wireless communication services. The proposed scheme results in a slight degradation in bit error rate performance for large block sizes because the effective interleaver size in a radix-4 implementation is reduced to half, relative to the conventional method. To prove the latency reduction, we implemented the proposed scheme on a field-programmable gate array and compared its decoding speed with that of a conventional decoder. The results show an improvement of at least five fold for a single iteration of turbo decoding.

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Ring-Opening Polymerization of ʟ-Lactide with Polydimethylsiloxane Based Stabilizers in Supercritical Carbon Dioxide (폴리디메틸실록산계 안정화제를 이용한 초임계 이산화탄소에서의 ʟ-Lactide의 개환중합)

  • Hwang, Ha Soo;Lim, Kwon Taek
    • Clean Technology
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    • v.12 no.2
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    • pp.62-66
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    • 2006
  • Poly($\small{L}$-Lactide)(PLLA) was prepared by a ring-opening polymerization of $\small{L}$-Lactide with various polydimethylsiloxane(PDMS) based copolymers as a stabilizer in supercritical carbon dioxide($scCO_2$). The block copolymeric stabilizers were synthesized by group transfer polymerization (GTP) by using PDMS macroinitiator. PLLA was found to be produced with fairly low molecular weight distribution as confirmed by gel permeation chromatography(GPC) analysis. Scanning electron microscopy (SEM) results showed that sub-micron size Poly($\small{L}$-lactide)(PLLA) particles were formed by suspension polymerization.

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