• KSBB Journal
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• 제31권4호
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• pp.284-290
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• 2016

In this research, recombinant human cytotoxic T-lymphocyte antigen 4-immunoglobulin (hCTLA4Ig) was produced by transgenic rice cells. RAmy3D promoter was used for overcome the limitation of low expression level in transgenic plant cells, and the secretion of target protein was accomplished by signal peptide. However, the RAmy3D promoter system which can be induced only by sugar starvation causes the decrease of cell viability. As a result, cell death promotes the release of protease which degrades the target proteins. The protein stability and productivity can be significantly influenced by proteolysis activity. Therefore, development of new strategies are necessary for the in situ recovery of target proteins from cell culture media. In this study, in situ recovery was performed by various strategies. Direct addition of Protein A resin with nylon bag leads to cell death by increased shear stress and decrease in production of hCTLA4Ig by protease. Medium exchange through modified flask could recover hCTLA4Ig with high cell viability and low protease activity, on the other hand, the productivity was lower than that of control. When in situ recovery was conducted at day 7 after induction in air-lift bioreactor, 1.94-fold of hCTLA4Ig could be recovered compared to control culture without in situ recovery. Consequently, in situ recovery of hCTLA4Ig from transgenic rice cell culture could enhance productivity significantly and prevent degradation of target proteins effectively.

• 한국응용과학기술학회지
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• 제38권4호
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• pp.1010-1019
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• 2021

유전에 매장된 석유는 1차, 2차, 그리고 3차에 걸쳐 회수된다. 3차 회수로 분류된 공법 중 미생물을 이용한 원유 회수증진법은 크게 in-situ와 ex-situ 공법으로 나눌 수 있다. In-situ 공법은 미생물을 저류층에 직접 투입하여 미생물의 대사활동을 통한 원유의 회수를 유도하는 공법이다. 대사물질 중 바이오 계면활성제는 저류층 잔존 유분의 유동화에 큰 역할을 한다. 한편, ex-situ 공법은 외부에서 미생물의 대사물질을 추출하여 저류층에 투입하여 원유를 회수하는 방법이다. 두 가지 공법 모두 친환경적이지만, 보다 경제적인 in-situ 공법이 선호된다. 미생물을 이용한 원유 회수증진법은 오랜기간 검토되었음에도, 현재까지도 파일럿 규모에서 여전히 평가가 진행 중이다. 본 논문에서는, in-situ 공법에 적용가능한 미생물 중 박테리아의 종 특성과 대사물질에 대해서 살펴보았다.

• KSBB Journal
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• 제30권3호
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• pp.103-108
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• 2015

Production of foreign proteins by transgenic plant cell cultures has several advantages such as post-translational modification, low risk of product contamination and low-cost production and purification. However, target proteins are degraded by extracellular proteases existing in the media. A solution to this problem is the use of perfusion culture and ion exchange chromatography for the application of integrated bioprocess using in situ recovery. With this method, production of human granulocyte-macrophage colony-stimulating factor (hGM-CSF) was investigated in this study. First, optimization of cell concentration during the induction phase for the production of hGM-CSF was examined. As cell concentration increased, the level of hGM-CSF was decreased due to the presence of extracellular proteases. Induction using sugarfree media produced 33% more hGM-CSF. The effects of pH on the binding of hGM-CSF to cationic and anionic exchange resins were also investigated. In terms of stability, optimal pH was found to be 5~7. In the case of using buffer exchange when CM-Sepharose was used as a cationic exchange resin, optimal pH for binding was 4.8 and adsorption yield was 77%. When DEAE-Sepharose was used as an anionic exchange resin, it was 5.5 (74%). Without buffer exchange, optimal pH was 4.6 (84%). From these results, an integrated bioprocess using in situ recovery with simultaneous production and separation of foreign protein in transgenic plant cell suspension cultures was found to be feasible.

• Journal of Microbiology and Biotechnology
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• 제14권5호
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• pp.932-937
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• 2004

In situ extraction by organic solvent was studied in order to improve the recovery yield of hydrocarbon from the culture of Botryococcus braunii, a green colonial microalga. When the solvent mixture of octanol as an extractive solvent and n-octane as a biocompatible solvent was added to a two-phase column, the algal growth was seriously inhibited, even at a low concentration of polar octanol. Therefore, a two-stage cell-recycle extraction process was proposed to improve the contact area between the organic phase and the aqueous phase. The hydrocarbon recovery with in situ cell-recycle extraction showed a three-fold increase (57% of cell) in yield over that with two-phase extraction. In addition, over 60% of the hydrocarbon could be recovered without serious cell damage by downstream separation when this process was applied to the culture broth after batch fermentation.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2002년도 생물공학의 동향 (X)
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• pp.321-324
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• 2002

본 연구에서는 hGM-CSF를 생산해 배지 내로 분바하도록 유전자 조작된 Nicotiana tabacum 세포를 수성이상계 내에서 배양하여 in situ recovery를 시도하였다. 특히 식물세포 자체의 생상에 큰 영향을 주지 않으면서도 일반 배지에서 회수한 단백질에 비해 많은 양을 생산할 수 있는 수성이상계 시스템을 선정하였다. 6% (w/w) PEG 20,000과 10% (w/w) dextran 2.000,000 을 이용하는 경우 최대 세포 농도가 18.6 g/L 로, 일반배지를 사용하는 경우 (15.7 g/L)에 비해 저해가 없음을 확인하였다. 또한 목적 단백질인 hGM-CSF의 생산에 있어서도 위의 시스템의 경우 dextran-rich phase인 아랫 상으로 분배됨으로 인해 회수가 쉬울 것으로 확인되었으며 그 생산량 또한 일반배지에서의 생산량 (1.5 ng/mL)에 비해 크게 다르지 않음을 확인할 수 있었다. 따라서 hGM-CSF의 회수를 위한 in situ recovery 시스템에 있어서 수성이상계의 이용 가능성을 확인하였다.

• 한국미생물·생명공학회지
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• 제23권5호
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• pp.609-616
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• 1995

Effects of lactate concentration, temperature, counter ions, pH as well as voltage (current) in batch electrodialysis (ED) experiments with a 3-compartment unit were investigated. The applied voltage was found to be the most critical factor as expected. The electrodialysis rate increased with the lactate concentration of the source solution. The amount of lactate transferred was limited by the lactate concentration difference between cathode and permeate compartments. The electrodialysis rate did not heavily depend on temperature change. The electrodialysis rate of NH$_{4}$-lactate was faster than that of Na-lactate and both lactates showed the highest electrodialysis rate at a pH of 4.0. A little amount of non-ionic glucose diffused through the anionic membrane to the permeate compartment. To test the effectiveness of the in situ recovery of lactic acid from fermentation broth by ED, three cases of batch culture were carried out; pH control only, ED only, and pH control and ED. The case with both pH control and ED was more efficient than that with pH control only in the aspects of productivity and product yield.

• Membrane and Water Treatment
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• 제6권3호
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• pp.205-224
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• 2015

A novel hydrophilic poly (vinylidene fluoride)/poly (p-phenylene terephthalamide) (PVDF/PPTA) blend membrane was prepared by in situ polycondensation of p-phenylene diamine (PPD) and terephthaloyl chloride (TPC) in PVDF solution with subsequent nonsolvent induced phase separation (NIPS) process. For comparison, conventional solution blend membrane was prepared directly by adding PVDF powder into PPTA polycondensation solution. Blend membranes were characterized by means of viscometry, X-ray photoelectron spectroscopy (XPS), Field Emission Scanning Electron Microscopy (FESEM). The effects of different blending methods on membrane performance including water contact angle (WCA), mechanical strength, anti-fouling and anti-compression properties were investigated and compared. Stronger interactions between PVDF and PPTA in in situ blend membranes were verified by viscosity and XPS analysis. The incorporation of PPTA accelerated the demixing rate and caused the formation of a more porous structure in blend membranes. In situ blend membranes exhibited better hydrophilicity and higher tensile strength. The optimal values of WCA and tensile strength were $65^{\circ}$ and 34.1 MPa, which were reduced by 26.1% and increased by 26.3% compared with pure PVDF membrane. Additionally, antifouling properties of in situ blend membranes were greatly improved than pure PVDF membrane with an increasing of flux recovery ratio by 25%. Excellent anti-compression properties were obtained in in situ blend membranes with a stable pore morphology. The correlations among membrane formation mechanism, structure and performance were also discussed.

• Geomechanics and Engineering
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• 제33권5호
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• pp.453-462
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• 2023

Intermediate Geomaterials (IGMs) are natural formation materials that exhibit the engineering behavior (strength and compressibility) between soils and rocks. The engineering behavior of such material is highly unpredictable as the IGMs are stiffer than soils and weaker/softer than rocks. Further, the characterization of such material needs exposure to both soil and rock mechanics. In most conventional designs of geotechnical structures, the engineering properties of the IGMs are either aligned with soils or rocks, and this assumption may end up either in an over-conservative design or under-conservative design. Hence, many researchers have attempted to evaluate its actual engineering properties through laboratory tests. However, the test results are partially reliable due to the poor core recovery of IGMs and the possible sample disturbance. Subsequently, in-situ tests have been used in recent years to evaluate the engineering properties of IGMs. However, the respective in-situ test finds its limitations while exploring IGMs with different geological formations at deeper depths with the constraints of sampling. Standard Penetration Test (SPT) is the strength-based index test that is often used to explore IGMs. Moreover, it was also observed that the coefficient of variation of the design parameters (which represents the uncertainties in the design parameters) of IGMs is relatively high, and also the studies on the probabilistic characterization of IGMs are limited compared with soils and rocks. With this perspective, the present article reviews the laboratory and in-situ tests used to characterize the IGMs and explores the shear strength variation based on their geological origin.

• Molecules and Cells
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• 제23권3호
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• pp.405-409
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• 2007

A putative type-I chalcone isomerase (CHI) cDNA clone EuNOD-CHI was previously isolated from the root nodule of Elaeagnus umbellata [Kim et al. (2003)]. To see if it encodes a functional CHI, we ectopically overexpressed it in the Arabidopsis (Arabidopsis thaliana) transparent testa 5 (tt5) mutant, which is defective in naringenin production and has yellow seeds due to proanthocyanidin deficiency. Ectopic overexpression of EuNOD-CHI resulted in recovery of normal seed coat color. Naringenin produced by CHI from naringenin chalcone was detected in the transgenic lines like in the wild-type, whereas it was absent from the tt5 mutant. We conclude that EuNOD-CHI encodes a functional type-I CHI. In situ hybridization revealed that EuNOD-CHI expression is localized to the infected cells of the fixation zone in root nodules.

• 반도체디스플레이기술학회지
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• 제19권1호
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• pp.61-66
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• 2020

Threshold voltage shift occurring during operation is implemented in a SPICE simulation tool. Among the shift models the stretched-exponential function model, which is frequently observed from both single-crystal silicon and thin-film transistors regardless of the nature of causes, is selected, adapted to transient simulation, and added to BSIM4 developed by BSIM Research Group at the University of California, Berkeley. The adaptation method used in this research is to select degradation and recovery models based on the comparison between the gate and threshold voltages. The threshold voltage shift is extracted from SPICE transient simulation and shows the stretched-exponential time dependence for both degradation and recovery situations. The implementation method developed in this research is not limited to the stretched-exponential function model and BSIM model. The proposed method enables to perform transient simulation with threshold voltage shift in situ and will help to verify the reliability of a circuit.