• Title/Summary/Keyword: optimization of production

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Characterization of Protease Produced by Elizabethkingia meningoseptica CS2-1 and Optimization of Cultural Conditions for Amino Acid Production (닭 우모 분해세균 Elizabethkingia meningoseptica CS2-1이 생산하는 단백질분해효소의 특성 및 아미노산 생산을 위한 배양조건)

  • Kim, Se-Jong;Cho, Chun-Hwi;Whang, Kyung-Sook
    • Journal of Applied Biological Chemistry
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    • v.54 no.2
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    • pp.135-142
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    • 2011
  • A feather-degrading bacterium Elizabethkingia meningoseptica CS2-1 was isolated from compost in a chicken farm. Cultured on a basal medium containing 2% chicken feather, the bacterium showed 729.7 ${\mu}mol/mL$ of amino acid. Optimal culture conditions for feather degradation by E. meningoseptica CS2-1 were $25^{\circ}C$, pH 7.5, and 180 rpm. The optimal pH and temperature for protease activity were 8.0 and $40^{\circ}C$, respectively. The composition of an optimal medium for amino acid production was 0.05% NH4Cl, 0.05% NaCl, 0.03% $K_2HPO_4$, 0.03% $KH_2PO_4$, 0.01% $MgCl_2{\cdot}6H_2O$, 0.1% urea, and 2% chicken feather. Characteristics of amino acids extracted from the optimal medium under the optimal culture conditions of E. meningoseptica CS2-1 were analyzed. The total amino acid content of strain CS2-1 was 1063 ${\mu}mol/mL$, which was 46% higher compared to the basal condition (729.7 ${\mu}mol/mL$). The essential amino acid content in the total amino acid was 315.9 ${\mu}mol/mL$, which was 44% higher than that of the basal condition. Major amino acids were proline (14%), aspartic acid (12%), glutamic acid (11%), serine (10%), alanine (10%), glycine (9%), and tyrosine (7%) by strain CS2-1. These results suggest that strain CS2-1 can be used as a potential microbial resource for the production of amino acid using chicken feathers.

Numerical and Experimental Study on the Coal Reaction in an Entrained Flow Gasifier (습식분류층 석탄가스화기 수치해석 및 실험적 연구)

  • Kim, Hey-Suk;Choi, Seung-Hee;Hwang, Min-Jung;Song, Woo-Young;Shin, Mi-Soo;Jang, Dong-Soon;Yun, Sang-June;Choi, Young-Chan;Lee, Gae-Goo
    • Journal of Korean Society of Environmental Engineers
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    • v.32 no.2
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    • pp.165-174
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    • 2010
  • The numerical modeling of a coal gasification reaction occurring in an entrained flow coal gasifier is presented in this study. The purposes of this study are to develop a reliable evaluation method of coal gasifier not only for the basic design but also further system operation optimization using a CFD(Computational Fluid Dynamics) method. The coal gasification reaction consists of a series of reaction processes such as water evaporation, coal devolatilization, heterogeneous char reactions, and coal-off gaseous reaction in two-phase, turbulent and radiation participating media. Both numerical and experimental studies are made for the 1.0 ton/day entrained flow coal gasifier installed in the Korea Institute of Energy Research (KIER). The comprehensive computer program in this study is made basically using commercial CFD program by implementing several subroutines necessary for gasification process, which include Eddy-Breakup model together with the harmonic mean approach for turbulent reaction. Further Lagrangian approach in particle trajectory is adopted with the consideration of turbulent effect caused by the non-linearity of drag force, etc. The program developed is successfully evaluated against experimental data such as profiles of temperature and gaseous species concentration together with the cold gas efficiency. Further intensive investigation has been made in terms of the size distribution of pulverized coal particle, the slurry concentration, and the design parameters of gasifier. These parameters considered in this study are compared and evaluated each other through the calculated syngas production rate and cold gas efficiency, appearing to directly affect gasification performance. Considering the complexity of entrained coal gasification, even if the results of this study looks physically reasonable and consistent in parametric study, more efforts of elaborating modeling together with the systematic evaluation against experimental data are necessary for the development of an reliable design tool using CFD method.

Fabrication of a Partial Genome Microarray of the Methylotrophic Yeast Hansenula polymorpha: Optimization and Evaluation of Transcript Profiling

  • OH , KWAN-SEOK;KWON, OH-SUK;OH, YUN-WI;SOHN, MIN-JEONG;JUNG, SOON-GEE;KIM, YONG-KYUNG;KIM, MIN-GON;RHEE, SANG-KI;GERD GELLISSEN,;KANG, HYUN-AH
    • Journal of Microbiology and Biotechnology
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    • v.14 no.6
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    • pp.1239-1248
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    • 2004
  • The methylotrophic yeast Hansenula polymorpha has been extensively studied as a model organism for methanol metabolism and peroxisome biogenesis. Recently, this yeast has also attracted attention as a promising host organism for recombinant protein production. Here, we describe the fabrication and evaluation of a DNA chip spotted with 382 open reading frames (ORFs) of H. polymorpha. Each ORF was PCR-amplified using gene-specific primer sets, of which the forward primers had 5'-aminolink. The PCR products were printed in duplicate onto the aldehyde-coated slide glasses to link only the coding strands to the surface of the slide via covalent coupling between amine and aldehyde groups. With the partial genome DNA chip, we compared efficiency of direct and indirect cDNA target labeling methods, and found that the indirect method, using fluorescent-labeled dendrimers, generated a higher hybridization signal-to-noise ratio than the direct method, using cDNA targets labeled by incorporation of fluorescence-labeled nucIeotides during reverse transcription. In addition, to assess the quality of this DNA chip, we analyzed the expression profiles of H. polymorpha cells grown on different carbon sources, such as glucose and methanol, and also those of cells treated with the superoxide­generating drug, menadione. The profiles obtained showed a high-level induction of a set of ORFs involved in methanol metabolism and oxidative stress response in the presence of methanol and menadione, respectively. The results demonstrate the sensitivity and reliability of our arrays to analyze global gene expression changes of H. polymorpha under defined environmental conditions.

PLEIOTROPHIN EFFECTS ON BINDING AND SUBSEQUENT OSTEOGENESIS OF HUMAN MESENCHYMAL STEM CELLS (Pleiotrophin이 골수 줄기 세포의 부착 및 골형성에 미치는 효과에 대한 연구)

  • Yoon, Jung-Ho;Eune, Jung-Ju;Jang, Hyon-Seok;Rim, Jae-Suk;Lee, Eui-Seok;Kim, Dae-Sung;Kwon, Jong-Jin
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.28 no.2
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    • pp.111-117
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    • 2006
  • An area of current research is investigating the app1ication of human mesenchymal stem cells or hMSCs as a cell-based regenerative therapy. In order to achieve effective bone regeneration, appropriate matrices functioning as cell-carriers must be identified and optimized in terms of function, efficacy and biocompatibility. Two methods of approaching optimization of matrices are to facilitate adhesion of the donor hMSCs and furthermore to facilitate recruitment of host progenitor cells to osteoblastic differentiation. Pleiotrophin is an extracellular matrix protein that was first identified in developing rat brains and believed to be associated with developing neuronal pathways. A recent publication by Imai and colleagues demonstrated that transgenic mice with upregulated pleiotrophin expression developed a greater volume of cortical as well as cancellous bone. The proposed mechanism of action of pleiotrophin is demonstrated here. Through either environmental stresses and/or intracellular regulation, there is an increase in pleiotrophin production. The pleiotrophin is released extracellularly into areas requiring bone deposition. A receptor-mediated process recruits host osteoprogenitor cells into these areas. Therefore, the aim of our study was to investigate the osteoconductive properties of pleiotrophin. We wanted to determine if pleiotrophin coating facilitates cellular adhesion and furthermore if this has any effect on hMSCs derived bone formation in an animal model. The results showed a dose dependent response of cellular adhesion in fibronectin samples, and cellular adhesion was facilitated with increasing pleiotrophin concentrations. Histologic findings taken after 5 weeks implantation in SCID mouse showed no presence of bone formation with only a dense fibrous connective tissue. Possible explanations for the results of the osteogenesis assay include inappropriate cell loading.

Heat transfer analysis in sub-channels of rod bundle geometry with supercritical water

  • Shitsi, Edward;Debrah, Seth Kofi;Chabi, Silas;Arthur, Emmanuel Maurice;Baidoo, Isaac Kwasi
    • Nuclear Engineering and Technology
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    • v.54 no.3
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    • pp.842-848
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    • 2022
  • Parametric studies of heat transfer and fluid flow are very important research of interest because the design and operation of fluid flow and heat transfer systems are guided by these parametric studies. The safety of the system operation and system optimization can be determined by decreasing or increasing particular fluid flow and heat transfer parameter while keeping other parameters constant. The parameters that can be varied in order to determine safe and optimized system include system pressure, mass flow rate, heat flux and coolant inlet temperature among other parameters. The fluid flow and heat transfer systems can also be enhanced by the presence of or without the presence of particular effects including gravity effect among others. The advanced Generation IV reactors to be deployed for large electricity production, have proven to be more thermally efficient (approximately 45% thermal efficiency) than the current light water reactors with a thermal efficiency of approximately 33 ℃. SCWR is one of the Generation IV reactors intended for electricity generation. High Performance Light Water Reactor (HPLWR) is a SCWR type which is under consideration in this study. One-eighth of a proposed fuel assembly design for HPLWR consisting of 7 fuel/rod bundles with 9 coolant sub-channels was the geometry considered in this study to examine the effects of system pressure and mass flow rate on wall and fluid temperatures. Gravity effect on wall and fluid temperatures were also examined on this one-eighth fuel assembly geometry. Computational Fluid Dynamics (CFD) code, STAR-CCM+, was used to obtain the results of the numerical simulations. Based on the parametric analysis carried out, sub-channel 4 performed better in terms of heat transfer because temperatures predicted in sub-channel 9 (corner subchannel) were higher than the ones obtained in sub-channel 4 (central sub-channel). The influence of system mass flow rate, pressure and gravity seem similar in both sub-channels 4 and 9 with temperature distributions higher in sub-channel 9 than in sub-channel 4. In most of the cases considered, temperature distributions (for both fluid and wall) obtained at 25 MPa are higher than those obtained at 23 MPa, temperature distributions obtained at 601.2 kg/h are higher than those obtained at 561.2 kg/h, and temperature distributions obtained without gravity effect are higher than those obtained with gravity effect. The results show that effects of system pressure, mass flowrate and gravity on fluid flow and heat transfer are significant and therefore parametric studies need to be performed to determine safe and optimum operating conditions of fluid flow and heat transfer systems.

Expression of human lactoferrin N-lobe in Pichia pastoris and its antibacterial activity (Pichia pastoris에서 사람 락토페린 N-lobe의 발현과 항균활성)

  • Won, Su-Jin;Jo, Jae-Hyung;Kim, Seung-Hwan;Kwon, Hyuk-Jin;Lee, Hyune-Hwan
    • Korean Journal of Microbiology
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    • v.51 no.3
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    • pp.271-279
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    • 2015
  • Lactoferrin (LF) is a multifunctional, iron-binding glycoprotein found in physiological secretions of mammals. LF shows antibacterial, antiviral and antifungal activities. In the present study, a gene encoding the N-terminal lobe of human lactoferrin (hLF) was isolated, cloned and expressed in methylotrophic yeast, Pichia pastoris. The recombinant hLF-N (rhLF-N) protein was secreted into the culture medium at the level of $458{\mu}g/ml$ in 3 L fermentor. The size of purified hLF-N was estimated as 35 kDa when analyzed by SDS-PAGE and western blotting. The rhLF-N was further confirmed by immunodiffusion using the anti-hLF polyclonal antibody. The expression profile analysis by qRT-PCR showed that the relative mRNA expression of rhLF-N was maximal after 2-3 days of methanol induction and reduced gradually at 4 days. The purified rhLF-N showed broad antibacterial activities against the pathogens such as Staphylococcus aureus, E. coli, Pseudomonas aeruginosa, Burkholderia cepacia, and Salmonella typhimurium. However, rhLF-N showed relatively lower activity when compared to peptides derived from LF. In spite of this weak activity, the rhLF-N expressed in P. pastoris might be more advantageous for the industrial application, because rhLF-N is secreted into the culture medium and the production can also be increased by optimization of culture conditions.

Development of a Biofungicide Using a Mycoparasitic Fungus Simplicillium lamellicola BCP and Its Control Efficacy against Gray Mold Diseases of Tomato and Ginseng

  • Shin, Teak Soo;Yu, Nan Hee;Lee, Jaeho;Choi, Gyung Ja;Kim, Jin-Cheol;Shin, Chul Soo
    • The Plant Pathology Journal
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    • v.33 no.3
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    • pp.337-344
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    • 2017
  • To develop a commercial product using the mycoparasitic fungus Simplicillium lamellicola BCP, the scale-up of conidia production from a 5-l jar to a 5,000-l pilot bioreactor, optimization of the freeze-drying of the fermentation broth, and preparation of a wettable powder-type formulation were performed. Then, its disease control efficacy was evaluated against gray mold diseases of tomato and ginseng plants in field conditions. The final conidial yields of S. lamellicola BCP were $3.3{\times}10^9conidia/ml$ for a 5-l jar, $3.5{\times}10^9conidia/ml$ for a 500-l pilot vessel, and $3.1{\times}10^9conidia/ml$ for a 5,000-l pilot bioreactor. The conidial yield in the 5,000-l pilot bioreactor was comparable to that in the 5-l jar and 500-l pilot vessel. On the other hand, the highest conidial viability of 86% was obtained by the freeze-drying method using an additive combination of lactose, trehalose, soybean meal, and glycerin. Using the freeze-dried sample, a wettable powder-type formulation (active ingredient 10%; BCP-WP10) was prepared. A conidial viability of more than 50% was maintained in BCP-WP10 until 22 weeks for storage at $40^{\circ}C$. BCP-WP10 effectively suppressed the development of gray mold disease on tomato with control efficacies of 64.7% and 82.6% at 500- and 250-fold dilutions, respectively. It also reduced the incidence of gray mold on ginseng by 65.6% and 81.3% at 500- and 250-fold dilutions, respectively. The results indicated that the new microbial fungicide BCP-WP10 can be used widely to control gray mold diseases of various crops including tomato and ginseng.

Intraspecies Androgenesis in Mud Loach (Misgurnus mizolepis): I. Optimization of the Egg Inactivation and Haploid Androgenesis Using Transgene Marker (미꾸라지(Misgurnus mizolepis)의 웅성발생성 처녀생식: I. 형질전환 유전자 표지를 이용한 웅성발생성 반수체 유도의 최적화)

  • Nam Yoon-Kwon;Noh Choong-Hwan;Kim Dong-Soo
    • Journal of Aquaculture
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    • v.19 no.3
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    • pp.166-172
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    • 2006
  • Induced androgenesis, a form of artificial parthenogenesis is an important tool for the generation and use of genetically isogenic or clonal fish strain. An optimized protocol for the genetic inactivation of mud loach (Misgurnus mizolepis) oocytes (i.e. production of androgenetic haploid) was developed using UV-irradiation. Various dose levels of UV significantly affected the fertilizing capacity of the eggs, hatchability of embryos and incidence of haploidy. Based on the extensive examinations of treatment conditions on embryo viability and haploid incidence, the optimum dose level of UV irradiation was turned out to be $10,800ergs/mm^2$ with 56.9% of hatching success and 94.6% of haploidy. The overall yield of putative androgen under optimized treatment condition was more than 50% out of total eggs inseminated. The success of androgenetic reproduction of haploid genome was verified by flow cytometry and PCR amplification of transgene that is exclusive to either one of parental sexes. However, a small portion $(8\sim11%)$ of presumed androgenetic haploid larvae was proven to contain residual DNA fragment(s) from maternal parent.

Predicting blast-induced ground vibrations at limestone quarry from artificial neural network optimized by randomized and grid search cross-validation, and comparative analyses with blast vibration predictor models

  • Salman Ihsan;Shahab Saqib;Hafiz Muhammad Awais Rashid;Fawad S. Niazi;Mohsin Usman Qureshi
    • Geomechanics and Engineering
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    • v.35 no.2
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    • pp.121-133
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    • 2023
  • The demand for cement and limestone crushed materials has increased many folds due to the tremendous increase in construction activities in Pakistan during the past few decades. The number of cement production industries has increased correspondingly, and so the rock-blasting operations at the limestone quarry sites. However, the safety procedures warranted at these sites for the blast-induced ground vibrations (BIGV) have not been adequately developed and/or implemented. Proper prediction and monitoring of BIGV are necessary to ensure the safety of structures in the vicinity of these quarry sites. In this paper, an attempt has been made to predict BIGV using artificial neural network (ANN) at three selected limestone quarries of Pakistan. The ANN has been developed in Python using Keras with sequential model and dense layers. The hyper parameters and neurons in each of the activation layers has been optimized using randomized and grid search method. The input parameters for the model include distance, a maximum charge per delay (MCPD), depth of hole, burden, spacing, and number of blast holes, whereas, peak particle velocity (PPV) is taken as the only output parameter. A total of 110 blast vibrations datasets were recorded from three different limestone quarries. The dataset has been divided into 85% for neural network training, and 15% for testing of the network. A five-layer ANN is trained with Rectified Linear Unit (ReLU) activation function, Adam optimization algorithm with a learning rate of 0.001, and batch size of 32 with the topology of 6-32-32-256-1. The blast datasets were utilized to compare the performance of ANN, multivariate regression analysis (MVRA), and empirical predictors. The performance was evaluated using the coefficient of determination (R2), mean absolute error (MAE), mean squared error (MSE), mean absolute percentage error (MAPE), and root mean squared error (RMSE)for predicted and measured PPV. To determine the relative influence of each parameter on the PPV, sensitivity analyses were performed for all input parameters. The analyses reveal that ANN performs superior than MVRA and other empirical predictors, andthat83% PPV is affected by distance and MCPD while hole depth, number of blast holes, burden and spacing contribute for the remaining 17%. This research provides valuable insights into improving safety measures and ensuring the structural integrity of buildings near limestone quarry sites.

Optimization of Lipase-Catalyzed Production of Structured Lipids from Canola Oil Containing Similar Composition of Triacylglycerols to Cocoa Butter (Canola Oil로부터 코코아버터와 유사한 Triacylglycerol 조성을 가진 재구성지질의 효소적 합성 최적화 연구)

  • Moon, Jun-Hee;Lee, Jeung-Hee;Shin, Jung-Ah;Hong, Soon-Taek;Lee, Ki-Teak
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.40 no.10
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    • pp.1430-1437
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    • 2011
  • Synthesis conditions of cocoa butter equivalents were optimized using the response surface method (RSM) by interesterification of canola oil (Ca), palmitic ethyl ester (PEE), and stearic ethyl ester (StEE). The reaction was catalyzed by immobilized lipase (Lipozyme TLIM) from Thermomyces lanuginosa to produce structured lipids containing a composition of triacylglycerols similar to cocoa butter. Reaction conditions were optimized using D-optimal design with the three reaction factors of the substrate molar ratio of canola oil to palmitic ethyl ester and stearic ethyl ester (Ca : PEE : StEE=1:1:3, 1:1.66:5, 1:2:6, 1:2.33:7, 1:3:9, $X_1$), enzyme ratio (2~6%, $X_2$), and reaction time (30~270 min, $X_3$). The optimal conditions that minimized acyl-migration while maximizing 1-palmitoyl-2-oleoyl-3-stearoyl glycerol (POS), 1,3-distearoyl-2-oleoyl glycerol (SOS), and 1,3-dipalmitoyl-2-oleoyl glycerol (POP) were predicted, resulting in Ca : PEE : StEE=1:3:9, 6% of enzyme ratio, and 40 min of reaction time. The reaction product of structured lipids was synthesized again under the same conditions, showing 10.43 area% of acyl-migration, 25.31 area% of POS/PSO, 19.79 area% of SOS, and 11.22 area% of POP.