• Title/Summary/Keyword: pH-gradient

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Two-Step Fed-Batch Culture of Recombinant Escherichia coli for Production of Bacillus licheniformis Maltogenic Amylase

  • Kim, Myoung-Dong;Lee, Woo-Jong;Park, Kwan-Hwa;Rhee, Ki-Hyeong;Seo, Jin-Ho
    • Journal of Microbiology and Biotechnology
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    • v.12 no.2
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    • pp.273-278
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    • 2002
  • Two-step fed-batch fermentations were carried out to overproduce Bacillus licheniformis maltogenic amylase (BLMA) in recombinant Escherichia coli. The first step was to increase the cell mass by controlling the feeding of a glucose solution, while the second step was designed to improve the amylase expression efficiency by supplementing organic nitrogen sources. The linear gradient feeding method was successfully adopted to maintain the glucose concentration below 0.2 g/l during the fed-batch mode, as effectively minimizing acetic acid formation. When the dissolved oxygen (DO) level became limiting, an accumulation of acetic acid and drastic decrease in specific BLMA productivity were observed. Glucose and organic nitrogen sources consisting of yeast extract and casein hydrolysate were simultaneously supplied in the pH-stat mode to further increase the specific BLMA expression efficiency. An organic nitrogen source consisting of 200 g/1 yeast extract and 100 g/1 casein hydrolysate was found to be the best among the various combinations tested. The feeding of an organic nitrogen source in the second-step fed-batch period was highly beneficial in enhancing the BLMA production. The optimized two-step fed-batch culture resulted in 78 g/l maximum dry cell mass and 443 U/ml maximum BLMA activity, corresponding to 1.5-fold increase in the dry cell mass and 3.7-fold enhancement in BLMA production, compared with the simple fed-batch fermentation.

Characteristics of Micro Floc in a Rapid Mixing Step at Different Coagulant Dose (급속혼화공정에서 응집제 주입률에 따른 미세입자의 성장특성)

  • Jun, Hang-Bae;Park, Sang-Min;Park, Noh-Back;Jung, Kyung-Su
    • Journal of Korean Society of Water and Wastewater
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    • v.21 no.2
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    • pp.243-252
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    • 2007
  • Effects of alum dosage on the particle growth were investigated by monitoring particle counts in a rapid mixing process. Kaolin was used for turbid water sample and several other chemicals were added to adjust pH and ionic strength. The range of velocity gradient and mixing time applied for rapid mixing were $200{\sim}300sec^{-1}$ and 30~180 sec, respectively. Particle distribution in the synthetic water sample was close to the natural water where their turbidity was same. The number of particles in the range of $10.0{\sim}12.0{\mu}m$ increased rapidly with rapid mixing time at alum dose of 20mg/L, however, the number of $8.0{\sim}9.0{\mu}m$ particles increased at alum dose of 50mg/L. The number of $14.0{\sim}25.0{\mu}m$ particles at alum dose of 20mg/L was 10 times higher than them at alum dose of 50mg/L. Dominant particle growth was monitored at the lower alum dose than the optimum dose from a jar test at an extended rapid mixing time(about 120 sec). The number of $8.0{\sim}14.0{\mu}m$ particles was lower both at a higher alum doses and higher G values. At G value of $200sec^{-1}$ and at alum dose of 10-20mg/L, residual turbidity was lower as the mixing time increased. But at alum dose above 40mg/L and at same G value, lower residual turbidity occurred in a short rapid mixing time. Low residual turbidity at G value of $300sec^{-1}$ occurred both at lower alum doses and at shorter mixing time comparing to the results at G value of $200sec^{-1}$.

Temporal and spatial variations of vegetation in a riparian zone of South Korea

  • Park, Hyekyung;Kim, Jae Geun
    • Journal of Ecology and Environment
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    • v.44 no.2
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    • pp.62-71
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    • 2020
  • Understanding vegetation structure and the relationship with environmental factors has been crucial for restoration and conservation of riparian zones. In this study, we conducted field survey in a riparian zone of Namhan River in South Korea both before and after flooding in order to understand temporal and spatial variations of riparian vegetation. There were significant temporal and spatial variations in species composition, and distribution patterns of vegetation were different along a gradient of elevation above the water level. At low elevation, Zizania latifolia was dominant throughout the field survey periods, and Bidens frondosa began to grow late and dominated both in post-flooding 1 and 2. Prior to flooding, Scirpus radicans and Polygonum thunbergii were widely distributed at middle elevation, while Artemisia vulgaris, Phragmites australis, and Miscanthus sacchariflorus were dominant at high elevation. After flooding, P. thunbergii was dominant at middle elevation with most other species decreasing, and more invasive or pioneer plants, including Artemisia princeps, H. scandens, and Sicyos angulatus, were observed at high elevation. Species composition and distribution patterns were homogeneous at low elevation, whereas dynamic variations of vegetation were observed both temporally and spatially at higher elevations. Elevation and distance from the water front were the most principal factors governing vegetation structure. Furthermore, soil physicochemical properties were also found to determine species composition and distribution patterns. These results indicate that vegetation structure in the riparian zones is formed by the combined effects of hydrological regime and soil physicochemical properties, inherent characteristics of species, and interspecific competition. Understanding of temporal and spatial variations of riparian vegetation may provide useful insights into ecological restoration and conservation of the vegetation within the riparian zones.

Magnetic-activated cell sorting improves high-quality spermatozoa in bovine semen

  • de Assumpcao, Teresinha Ines;Severo, Neimar Correa;Zandonaide, Joao Pedro Brandao;Macedo, Gustavo Guerino
    • Journal of Animal Reproduction and Biotechnology
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    • v.36 no.2
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    • pp.91-98
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    • 2021
  • The objective of this study was to establish a selection process for high quality sperm in bovine semen using sperm separation by magnetic activation (MACS). For this, semen from 21 Nellore bulls was collected using an artificial vagina. To guarantee the presence of pathologies in the ejaculate, animals previously declassified in four consecutive spermiogram were used. Semen was analyzed in five statuses: (1) fresh semen (fresh); (2) density gradient centrifugation (DGC), percoll column; (3) non-apoptotic fraction after separation by MACS (MAC); (4) apoptotic fraction from the separation (MACPOOR); and (5) MAC followed by DGC (MACDGC). Using a computerized analysis system (CASA), motility was measured. The sperm morphology was evaluated by phase contrast, and the supravital test was completed with eosin/nigrosin staining. For DGC, 20 × 106 cells were used in a gradient of 90% and 45% percoll. MACS used 10 × 106 cells with 20 μL of nanoparticles attached to annexin V, and filtered through the MiniMACS magnetic separation column. Membrane integrity was assessed with SYBR-14/IP and mitochondrial potential with JC-1 by flow cytometry. Processing sperm by MACDGC, was more effective in obtaining samples with high quality sperm, verified by the total of abnormalities in the samples: 35.04 ± 2.29%, 21.50 ± 1.47%, 17.30 ± 1.10%, 30.68 ± 1.94% and 10.50 ± 1.46%, respectively for fresh, DGC, MAC, MACPOOR, and MACDGC. The subpopulation of non-apoptotic sperm had a high number of live cells (82.65%), membrane integrity (56.60%) and mitochondrial potential (83.98%) (p < 0.05). These findings suggest that this nanotechnological method, that uses nanoparticles, is efficient in the production of high-quality semen samples for assisted reproduction procedures in cattle.

A Study on Data-driven Modeling Employing Stratification-related Physical Variables for Reservoir Water Quality Prediction (취수원 수질예측을 위한 성층 물리변수 활용 데이터 기반 모델링 연구)

  • Hyeon June Jang;Ji Young Jung;Kyung Won Joo;Choong Sung Yi;Sung Hoon Kim
    • Proceedings of the Korea Water Resources Association Conference
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    • 2023.05a
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    • pp.143-143
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    • 2023
  • 최근 대청댐('17), 평림댐('19) 등 광역 취수원에서 망간의 먹는 물 수질기준(0.05mg/L 이하) 초과 사례가 발생되어, 다수의 민원이 제기되는 등 취수원의 망간 관리 중요성이 부각되고 있다. 특히, 동절기 전도(Turn-over)시기에 고농도 망간이 발생되는 경우가 많은데, 현재 정수장에서는 망간을 처리하기 위해 유입구간에 필터를 설치하고 주기적으로 교체하는 방식으로 처리하고 있다. 그러나 단기간에 고농도 망간 다량 유입 시 처리용량의 한계 등 정수장에서의 공정관리가 어려워지므로 사전 예측에 의한 대응 체계 고도화가 필요한 실정이다. 본 연구는 광역취수원인 주암댐을 대상으로 망간 예측의 정확도 향상 및 예측기간 확대를 위해 다양한 머신러닝 기법들을 적용하여 비교 분석하였으며, 독립변수 및 초매개변수 최적화를 진행하여 모형의 정확도를 개선하였다. 머신러닝 모형은 수심별 탁도, 저수위, pH, 수온, 전기전도도, DO, 클로로필-a, 기상, 수문 자료 등의 독립변수와 화순정수장에 유입된 망간 농도를 종속변수로 각 변수에 해당하는 실측치를 학습데이터로 사용하였다. 그리고 데이터기반 모형의 정확도를 개선하기 위해서 성층의 수준을 판별하는 지표로서 PEA(Potential Energy Anomaly)를 도입하여 데이터 분석에 활용하고자 하였다. 분석 결과, 망간 유입률은 계절 주기에 따라 농도가 달라지는 것을 확인하였고 동절기 전도시점과 하절기 장마기간 난류생성 시기에 저층의 고농도 망간이 유입이 되는 것을 분석하였다. 또한, 두 시기의 망간 농도의 변화 패턴이 상이하므로 예측 모델은 각 계절별로 구축해 학습을 진행함으로써 예측의 정확도를 향상할 수 있었다. 다양한 머신러닝 모델을 구축하여 성능 비교를 진행한 결과, 동절기에는 Gradient Boosting Machine, 하절기에는 eXtreme Gradient Boosting의 기법이 우수하여 추론 모델로 활용하고자 하였다. 선정 모델을 통한 단기 수질예측 결과, 전도현상 발생 시기에 대한 추종 및 예측력이 기존의 데이터 모형만 적용했을 경우대비 약 15% 이상 예측 효율이 향상된 것으로 나타났다. 본 연구는 머신러닝 모델을 활용한 망간 농도 예측으로 정수장의 신속한 대응 체계 마련을 지원하고, 수처리 공정의 효율성을 높이는 데 기여할 것으로 기대되며, 후속 연구로 과거 시계열 자료 활용 및 물리모형과의 연결 등을 통해 모델의 신뢰성을 제고 할 계획이다.

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Rates of Recovery and of Development In vitro of Follicular Oocytes Collected by Aspiration and Slicing Method in Cattle (채취 방법에 따른 소 난포란의 회수율 및 수정란의 발달율)

  • 조상래;강태영;박종식;허창기;송상현;이효종;최상용
    • Journal of Embryo Transfer
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    • v.16 no.2
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    • pp.99-106
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    • 2001
  • This study was undertaken to compare the efficiency of recovery rate and development rate of follicular oocytes collected either by aspiration or by slicing method. The follicular oocytes collected by the two methods matured in TCM199 supplemented with 10% steer serum at 39$^{\circ}C$ in a humidified atmosphere of 5% $CO_2$in air. After 22 h of culture, the oocytes were inseminated with frozen-thawed semen (2$\times$10$^{6}$ sperm/ml of final concentration) prepared with Percoll-density gradient in IVF-TALP medium for 16 h. Later, sets of 15 presumptive zygotes were transferred into 50 $\mu$L, droplets of CR1aa medium. On day 4 of the culture, embryos were transferred to TCM199 until day 9. The percentages of nuclear maturation to pre-metaphase II in the oocytes collected by aspiration are significantly (P<0.05) higher than that by slicing (83% vs. 62%, respectively). The mean number of oocytes recovered by slicing per ovary is significantly (P<0.05) higher than that by aspiration (15.1 vs. 6.7, respectively). Although the rates of cleavage and development to blastocyst of oocytes collected b)\\\\`aspiration are significantly (P<0.05) higher than that by slicing, the number of transferable embryos obtained by slicing method is significantly (P<0.05) higher than that by aspiration. From the results. we may conclude that slicing method is better than aspiration method for obtaining large number of transferable embryos per ovary.

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$KTiOPO_4 (KTP)$ Single Crystal Growth by TSSG Technique (TSSG법에 의한 $KTiOPO_4 (KTP)$ 단결정 육성)

  • 김정환;강진기
    • Korean Journal of Crystallography
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    • v.3 no.1
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    • pp.37-43
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    • 1992
  • KTiOP04 is a nonlinear optical crystal which is most widely used for frequency doubling of the radiation of Nd : YAG laser. In the experiment, sin ale crystals of KTiOP04 were grown by TSSG technique using 3K2W04·P2O5 flux. Low temperature gradient furnace suitable for KTP single crystal growth was used. Seed crystal was placed at the surface of the solution for the purpose of better observation of the growing crystals and the possibility of diameter control. Solution included 66.7mol% KTiOP04 for all experiments and its saturation temperature was 1020℃. The conditions of single crystal growth were as follows: cooling rate 0.2℃/h, crystal rotation rate 50rpm, c -axis seed. Using these conditions, single crystals up to 23 ×25×25mm3 have been groan from about 100cc solution. We have also observed a change in the crystal growth habit which resulted in the formation of large (201) faces and small (100) faces. And some crystals have (101) faces.

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Effect of Nifedipine on the Ampicillin Absorption (니페디핀이 암피실린의 흡수에 미치는 영향)

  • Jeong, Hyun-Jeong;Yong, Chul-Soon;Choi, Yoon-Soo;Oh, Doo-Man
    • Journal of Pharmaceutical Investigation
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    • v.27 no.1
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    • pp.57-64
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    • 1997
  • $Amino-{\beta}-lactam$ antibiotics are absorbed by the dipeptide transporter in the small intestine. These uptakes are coupled to a proton influx. The inward proton gradient is partly induced by the $Na^+/H^+$ exchanger and calcium ion is involved in control of this antiport. Interaction between ampicillin which is one of the $Amino-{\beta}-lactam$ antibiotics and nifedipine which is one of calcium channel blocking agents was studied in rats in vivo and with rabbit jejunum mounted on the Sweetana/Grass diffusion cells in vitro. Bioavailability of ampicillin was increased significantly when nifedipine was co-administered orally in rats. There were no differences in the distribution phase and the elimination phase when ampicillin was given either alone or with nifedipine intravenously. Conditions for in vitro experiments were determined. The lift rate of $O_2/CO_2$ gas was controlled to 3 bubbles/sec and ampicillin was stable in the Kreb's buffer at pH 6.0. Absorption of ampicillin was the greatest when the completely-stripped serosal membrane was used. Transport of ampicillin from mucosal to serosal side in the rabbit jejunum was enhanced by 32% in the presence of nifedipine (p=0.059). Above results suggest that nifedipine might increase the plasma level of ampicillin via the improved absorption in the intestine rather than the reduction in the elimination or/and alteration in the distribution.

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Effect of Low Salt Concentrations on Microbial Changes During Kimchi Fermentation Monitored by PCR-DGGE and Their Sensory Acceptance

  • Ahmadsah, Lenny S. F.;Min, Sung-Gi;Han, Seon-Kyeong;Hong, Yeun;Kim, Hae-Yeong
    • Journal of Microbiology and Biotechnology
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    • v.25 no.12
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    • pp.2049-2057
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    • 2015
  • Various salt concentrations (1.0%, 1.3%, 1.6%, 1.9%, and 2.1% labeled as sample A, B, C, D, and E, respectively) were investigated for microbial diversity, identification of Lactic Acid Bacteria (LAB) in salted kimchi cabbage, prepared under laboratory conditions. These samples were stored at 4°C for 5 weeks in proper aluminum-metalized pouch packaging with calcium hydroxide gas absorber. A culture-independent method known as polymerase chain reaction - denaturing gradient gel electrophoresis was carried out to identify LAB distributions among various salt concentration samples that had identified 2 Weissella (W. confusa and W. soli), 1 Lactobacillus (Lb. sakei), and 3 Leuconostoc (Lc. mesenteroides, Lc. lactis, and Lc. gelidum) in the overall kimchi samples. The pH, titratable acidity, viable cell counts, and coliform counts were not affected by salt variations. In order to assess sensory acceptance, the conducted sensory evaluation using a 9-point hedonic scale had revealed that samples with 1.3% salt concentration (lower than the manufacturer's regular salt concentration) was more preferred, indicating that the use of 1.3% salt concentration was acceptable in normal kimchi fermentation for its quality and safety. Despite similarities in pH, titratable acidity, viable cell counts, coliform counts, and LAB distributions among the various salt concentrations of kimchi samples, the sample with 1.3% salt concentration was shown to be the most preferred, indicating that this salt concentration was suitable in kimchi production in order to reduce salt intake through kimchi consumptions.

Separation of Glutathione by Ion Exchange Chromatography (이온교환 크로마토그래픽을 이용한 Glutathione 분리)

  • 김정훈;손영종;구윤모
    • KSBB Journal
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    • v.13 no.1
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    • pp.77-82
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    • 1998
  • Glutathione(L-$\gamma$ -glutamyl-L-cysteinylglycine, GSH) produced by microbial enzymes was separated by a liquid chromatography. In order to select a resin which would bind GSH efficiently, a batch adsorption experiment was carried out with GSH solution and various resins at pH 8.0 GSH bound to Q-sepharose and QAE-sephadex among anion exchange resins, but the latter was found not to be suitable because of the reduction of resin volume at high salt concentration. Preliminary experiments using a standard solution were carried out to separate GSH. GSH and $\gamma$ -glutamylcysteine were separated from the other constituents by applying step gradient of salt(NaCl) concentration. GSH was successfully separated from $\gamma$ -glutamylcysteine by applying Tris buffer containing 35mM NaCl. Chromatographic separation behaviors for the enzymatic product was similar to that for the standard solution. Separation yields of GSH from the standard solution and enzymatic product solution were 72.6% and 84.4%, respectively.

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