• 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.

• Biotechnology and Bioprocess Engineering:BBE
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• 제11권6호
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• pp.510-515
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• 2006

The putative EPA synthesis gene cluster was mined from the entire genome sequence of Shewanella oneidensis MR-1. The gene cluster encodes a PKS-like pathway that consists of six open reading frames (ORFs): ORFSO1602 (multi-domain beta-ketoacyl synthase, KS-MAT-4ACPs-KR), ORFSO1600 (acyl transferase, AT), ORFSO1599 (multi-domain beta-ketoacyl synthase, KS-CLF-DH-DH), ORFSO1597 (enoyl reductase, ER), ORFSO1604 (phosphopentetheine transferase, PPT), and ORFSO1603 (transcriptional regulator). In order to prove involvement of the PKS-like machinery in EPA synthesis, a 20.195-kb DNA fragment containing the genes was amplified from S. oneidensis MR-1 by the long-PCR method. Its identity was confirmed by the methods of restriction enzyme site mapping and nested PCR of internal genes orfSO1597 and orfSO1604. The DNA fragment was cloned into Escherichia coli using cosmid vector SuperCos1 to form pCosEPA. Synthesis of EPA was observed in four E. coli clones harboring pCosEPA, of which the maximum yield was 0.689% of the total fatty acids in a clone designated 9704-23. The production yield of EPA in the E. coli clone was affected by cultivation temperature, showing maximum yield at $20^{\circ}C$ and no production at $30^{\circ}C$ or higher. In addition, production yield was inversely proportional to glucose concentration of the cultivation medium. From the above results, it was concluded that the PKS-like modules catalyze the synthesis of EPA. The synthetic process appears to be subject to regulatory mechanisms triggered by various environmental factors. This most likely occurs via the control of gene expression, protein stability, or enzyme activity.

• Journal of Microbiology and Biotechnology
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• 제32권8호
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• pp.1041-1046
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• 2022

Nucleoside deoxyribosyltransferase (NDT) is an enzyme that replaces the purine or pyrimidine base of 2'-deoxyribonucleoside. This enzyme is generally used in the nucleotide salvage pathway in vivo and synthesizes many nucleoside analogs in vitro for various biotechnological purposes. Since NDT is known to exhibit relatively low reactivity toward nucleoside analogs such as 2'-fluoro-2'-deoxynucleoside, it is necessary to develop an enhanced NDT mutant enzyme suitable for nucleoside analogs. In this study, molecular evolution strategy via error-prone PCR was performed with ndt gene derived from Lactobacillus leichmannii as a template to obtain an engineered NDT with higher substrate specificity to 2FDU (2'-fluoro-2'-deoxyuridine). A mutant library of 214 ndt genes with different sequences was obtained and performed for the conversion of 2FDU to 2FDA (2'-fluoro-2'-deoxyadenosine). The E. coli containing a mutant NDT, named NDT^L59Q, showed 1.7-fold (at 40℃) and 4.4-fold (at 50℃) higher 2FDU-to-2FDA conversions compared to the NDT^WT, respectively. Subsequently, both NDT^WT and NDT^L59Q enzymes were over-expressed and purified using a His-tag system in E. coli. Characterization and enzyme kinetics revealed that the NDT^L59Q mutant enzyme containing a single point mutation of leucine to glutamine at the 59^th position exhibited superior thermal stability with enhanced substrate specificity to 2FDU.

• Journal of Electrochemical Science and Technology
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• 제15권1호
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• pp.198-206
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• 2024

Given the high theoretical capacity (1,675 mAh g^-1) and the inherent affordability and ubiquity of elemental sulfur, it stands out as a prominent cathode material for advanced lithium metal batteries. Traditionally, sulfur was sequestered within conductive porous carbons, rooted in the understanding that their inherent conductivity could offset sulfur's non-conductive nature. This study, however, pivots toward a transformative approach by utilizing diatom shell (DS, diatomite)-a naturally abundant and economically viable siliceous mineral-as a sulfur host. This approach enabled the development of a sulfurlayered diatomite/S composite (DS/S) for cathodic applications. Even in the face of the insulating nature of both diatomite and sulfur, the DS/S composite displayed vigorous participation in the electrochemical conversion process. Furthermore, this composite substantially curbed the loss of soluble polysulfides and minimized structural wear during cycling. As a testament to its efficacy, our Li-S battery, integrating this composite, exhibited an excellent cycling performance: a specific capacity of 732 mAh g^-1 after 100 cycles and a robust 77% capacity retention. These findings challenge the erstwhile conviction of requiring a conductive host for sulfur. Owing to diatomite's hierarchical porous architecture, eco-friendliness, and accessibility, the DS/S electrode boasts optimal sulfur utilization, elevated specific capacity, enhanced rate capabilities at intensified C rates, and steadfast cycling stability that underscore its vast commercial promise.

• 한국산림과학회지
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• 제100권1호
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• pp.112-117
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• 2011

본 연구는 산불이 저서성대형무척추동물의 섭식기능군과 군집안정성에 미치는 영향을 파악하기 위해 실시 되었다. 산불이 발생한 울진군의 2개 수계를 실험수계 그리고 산불비발생지의 2개 수계를 대조수계로 설정하여 산불이 발생한 2007년부터 2009년까지 조사하였다. 조사결과, 대조수계에서 4문 6강 16목 47과 89종, 실험수계에서 4문 6강 16목 43과 84종이 출현하였다. 각 연도별 실험수계에서 조사된 청정수계지표종인 E.P.T 분류군의 종수 및 개체수의 경우, 2007년에는 하루살이목이 21종, $2,434.6inds./m^2$, 강도래목이 3종, $199.8inds./m^2$, 날도래목이 14종, $540.2inds./m^2$로 나타났으며, 2009년에는 하루살이목 9종, $296inds./m^2$, 날도래목 4종 $44.4inds./m^2$, 강도래목은 나타나지 않았다. 전체적으로 매년 E.P.T 분류군에 속하는 종수 및 개체수가 급감하는 것으로 나타났다. 또한 매년 대조수계에 비해 실험수계에서 다양도와 풍부도가 감소하는 경향을 보였다. 섭식기능군은 GC type은 종수 및 개체수가 매년 감소하는 경향을 나타내었으나 기타 type에는 큰 변화가 없었다. 군집안정성은 실험수계에서 I III구역에 해당하는 종이 가장 많이 출현하였으나, 산불 후 매년 I III구역의 종수가 대체적으로 감소하는 경향을 나타내었다.

• 보존과학회지
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• 제20권
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• pp.43-54
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• 2007

무량사오층석탑(보물 제185호)은 백제의 옛 땅에 위치한 지리적 특성으로 백제의 기법이 이어졌고 통일신라의 시대적인 양식도 계승된 고려초기의 석탑이다. 이 석탑은 오랜세월 외부에 위치해 있어 부재에 심한 암석학적 풍화와 생물학적 풍화를 받고 있으며 상대갑석에는 중심침하가 발생한 상태이다. 이에 상대갑석의 중심침하의 원인을 규명하기 위해 탄성파 탐사, 전기비저항 탐사, GPR 탐사와 같은 비파괴 방법을 이용하여 지반안정해석을 실시하였다. 이 결과, 모든 탐사에서 지반은 양호한 상태로 나타났으며, 특히 전기비저항 탐사자료를 통해 지반강화 다짐으로 보여지는 고비저항대가 나타났다. 따라서 상대갑석에서 발생한 중심침하 현상은 지반의 불안정에 의한 것이 아니라 상대갑석 상부 부재의 구조적 불안정에서 기인된 것이라 해석된다. 이 결과는 석탑의 상시 모니터링 또는 보존대책 수립시 기초자료로 활용될 것이다.

• 대한치과교정학회지
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• 제47권4호
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• pp.229-237
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• 2017

Objective: The aim of this study was to compare the initial stability as insertion and removal torque and the clinical applicability of novel orthodontic zirconia micro-implants made using a powder injection molding (PIM) technique with those parameters in conventional titanium micro-implants. Methods: Sixty zirconia and 60 titanium micro-implants of similar design (diameter, 1.6 mm; length, 8.0 mm) were inserted perpendicularly in solid polyurethane foam with varying densities of 20 pounds per cubic foot (pcf), 30 pcf, and 40 pcf. Primary stability was measured as maximum insertion torque (MIT) and maximum removal torque (MRT). To investigate clinical applicability, compressive and tensile forces were recorded at 0.01, 0.02, and 0.03 mm displacement of the implants at angles of $0^{\circ}$, $10^{\circ}$, $20^{\circ}$, $30^{\circ}$, and $40^{\circ}$. The biocompatibility of zirconia micro-implants was assessed via an experimental animal study. Results: There were no statistically significant differences between zirconia micro-implants and titanium alloy implants with regard to MIT, MRT, or the amount of movement in the angulated lateral displacement test. As angulation increased, the mean compressive and tensile forces required to displace both types of micro-implants increased substantially at all distances. The average bone-to-implant contact ratio of prototype zirconia micro-implants was $56.88{\pm}6.72%$. Conclusions: Zirconia micro-implants showed initial stability and clinical applicability for diverse orthodontic treatments comparable to that of titanium micro-implants under compressive and tensile forces.

• Journal of Microbiology and Biotechnology
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• 제15권1호
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• pp.29-34
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• 2005

W/O/W (water-in-oil-in-water) type multiple emulsion was applied to improve the storage stability of an antagonistic microorganism, Burkholderia gladioli. Encapsulation of microorganism into a W/O/W emulsion was conducted by using a two-step emulsification method. W/O/W emulsion was prepared by the incorporation of B. gladioli into rapeseed oil and the addition of polyglycerin polyriconolate (PGPR) and castor oil polyoxyethylene (COG 25) as the primary and secondary emulsifier, respectively. Microcrystalline cellulose was used as an emulsion stabilizer. To evaluate the usefulness of W/O/W emulsion formulation as a microbial pesticide for controlling the bacterial wilt pathogen (Ralstonia solanacearum), the storage stability and antagonistic activity of emulsion formulation were tested in vitro. The storage stability test revealed that the viability of formulated cells in emulsion was higher than that of unformulated cells in culture broth. At $4^{\circ}C$, the viabilities of formulated cells and unformulated cells at the end of 20 weeks decreased to about 2 and 5 log cycles, respectively. At $37^{\circ}C$, the viability of formulated cells decreased to only 2 log cycles at the end of storage. On the other hand, the viable cells in culture broth were not detected after 13 weeks. In activity test, formulated cells in emulsion were more effective in inhibiting the growth of pathogen than unformulated cells in culture broth. Unformulated cells completely lost their antagonistic activity during storage under similar conditions. The W/O/W multiple emulsion formulation was shown to be useful as the novel liquid formulation for biological control.

• 한국운동역학회지
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• 제27권1호
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• pp.45-52
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• 2017

Objective: The aim of this study was to investigate the periodicity of the lower extremity joint flexion/extension angle to compare the local stability between young and elderly women during walking on a treadmill. Method: Eighteen young women (mean $age=21.2{\pm}1.6y$; mean $mass=57.1{\pm}6.1kg$; mean $height=1.61{\pm}0.04m$) and 18 elderly women (mean $age=66.4{\pm}1.2y$; mean $mass=55.4{\pm}8.3kg$; mean $height=1.56{\pm}0.04m$) participated in this study. Approximate entropy (ApEn) was used to determine the periodicity in the lower limb joint angles. Results: The ApEn values of the two groups were statistically greater in the surrogate data test than in the original time series data (p<.05). The periodicity of the hip and ankle flexion/extension angles decreased in the elderly women group compared with the young women group (p<.05). The periodicity of the lower extremity joint flexion/extension angle showed that the ankle joint increased dominatingly in both groups (p<.05); the hip joint decreased compared with the knee joint in the young women group; and the knee joint decreased compared with the hip joint in the elderly women group (p<.05). Conclusion: These results suggest that the lower extremity joint flexion/extension angles of the young and elderly women during walking contained random noises as well as biological signals. In addition, the differences in the periodicity in the lower extremity joint between the young and elderly women may provide some insight in predicting potential falls and be used as a characteristic indicator for determining local stability in elderly women during walking.

• Preventive Nutrition and Food Science
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• 제21권3호
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• pp.289-295
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• 2016

Curcumin is a flavonoid found in the rhizome of the turmeric plant (Curcuma longa L.) and has recently attracted interest because it has numerous biological functions and therapeutic properties. In the present study, we attempted to incorporate curcumin into medium-chain triglyceride (MCT) nanoemulsions (0.15 wt% curcumin, 10 wt% MCT oil, and 10 wt% emulsifiers) with various emulsifiers [polyoxyethylene (20) sorbitan monolaurate (Tween-20), sorbitan monooleate (SM), and soy lecithin (SL)]. The physicochemical properties of the nanoemulsions including the Ostwald ripening stability were investigated. The initial droplet size was found to be 89.08 nm for the nanoemulsion with 10 wt% Tween-20 (control), and when Tween-20 was partially replaced with SM and SL, the size decreased: 73.43 nm with 4 wt% SM+6 wt% Tween-20 and 67.68 nm with 4 wt% SL+6 wt% Tween-20 (prepared at 15,000 psi). When the nanoemulsions were stored for 28 days at room temperature, the droplet size increased as the storage time increased. The largest increase was observed for the control nanoemulsion, followed by the 4 wt% SL+6 wt% Tween-20 and 4 wt% SM+6 wt% Tween-20 systems. The Turbiscan dispersion stability results strongly supported the relationship between droplet size and storage time. The time-dependent increase in droplet size was attributed to the Ostwald ripening phenomenon. Thus, the Ostwald ripening stability of curcumin-loaded MCT nanoemulsions with Tween-20 was considerably improved by partially replacing the Tween-20 with SM or SL. In addition, curcumin may have acted as an Ostwald ripening inhibitor.