• KSBB Journal
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• 제17권2호
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• pp.153-157
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• 2002

Poly-Iysine이 tagging된 hEGF와 angiogenin(6L10ESA)의 융합단백질의 고체상 재접힘이 heparin-Sepharose colullln에서 수행되었을 때, untagging 단백질(E5h)의 기존의 액상 재접힘 방법과 비교하여 재접힘 수율은 약 13배 정도 증가하였다. 게다가 poly-Iysine tagging된angiogenin은 heparin에 친화도를 높여주므로 2.5배에서 3배 정도의 흡탁 수율이 증가한다. 재접힘 수율은 고체상 반응으로 인해 높은 재현성을 보였다. 재접힘 공정시간은 대략 8배 단축되었다. 고체상 재전힘된 단백질은 자신의 생물학적 역가를 유지하였다. 따라서 이 연구는 고체상 재접힘 방법이 분자간의 상호작용을 억제하여 응집현상을 현저히 줄였기 때문에 기인한 결과로 생각된다. 따라서 응집으로 인한 재접힘 수율이 낮은 단백질의 재접힘 긍정에 고체상 재접힘 공정을 사용하면 높은 재접힘 수율을 얻을 수 있다.

• 정보처리학회논문지:소프트웨어 및 데이터공학
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• 제7권4호
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• pp.129-134
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• 2018

최근, 인공신경망 모델은 예측, 수치제어, 로봇제어, 패턴인식 등의 분야에서 촉망되는 기술이다. 본 연구에서는 인공신경망 모델을 이용하여 온실 외부 온도를 예측하고 이를 온실제어에 활용하는데 목적이 있다. 예측 모델의 성능 평가를 위해 다중회귀모델과 SVM 모델과의 비교분석을 수행하였다. 평가 방법으로는 10-Fold Cross Validation을 사용하였으며, 예측 성능 향상을 위해 상관관계분석 통해 데이터 축소를 수행하였고, 측정 데이터로부터 새로운 Factor 추출하여 데이터의 신뢰성을 확보하였다. 인공신경망 구축을 위해 Backpropagation algorithm을 사용하였으며, 다중회귀모델은 M5 method로 구축하였고, SVM 모델을 epsilon-SVM으로 구축하였다. 각 모델의 비교분석 결과 각각 0.9256, 1.8503과 7.5521로 나타났다. 또한 예측모델을 온실 난방부하 계산에 적용함으로써 온실에 사용되는 에너지 비용 절감을 통한 수입증대에 기여할 수 있다. 실험한 온실의 난방부하는 3326.4kcal/h이며, 총 난방시간이 $10000^{\circ}C/h$일 때 연료소비량은 453.8L로 예측된다. 아울러 데이터 마이닝 기술 중 하나인 인공신경망을 정밀온실제어, 재배기법, 수확예측 등 다양한 농업 분야에 적용함으로써 스마트 농업으로의 발전에 기여할 수 있다.

• International Journal of Fuzzy Logic and Intelligent Systems
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• 제15권4호
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• pp.268-276
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• 2015

A ship's sailing route or plan is determined by the master as the decision maker of the vessel, and depends on the characteristics of the navigational environment and the conditions of the ship. The trajectory, which appears as a result of the ship's navigation, is monitored and stored by a Vessel Traffic Service center, and is used for an analysis of the ship's navigational pattern and risk assessment within a particular area. However, such an analysis is performed in the same manner, despite the different navigational environments between coastal areas and the harbor limits. The navigational environment within the harbor limits changes rapidly owing to construction of the port facilities, dredging operations, and so on. In this study, a support vector machine was used for processing and modeling the trajectory data. A K-fold cross-validation and a grid search were used for selecting the optimal parameters. A complicated traffic route similar to the circumstances of the harbor limits was constructed for a validation of the model. A group of vessels was composed, each vessel of which was given various speed and course changes along a specified route. As a result of the machine learning, the optimal route and voyage data model were obtained. Finally, the model was presented to Vessel Traffic Service operators to detect any anomalous vessel behaviors. Using the proposed data modeling method, we intend to support the decision-making of Vessel Traffic Service operators in terms of navigational patterns and their characteristics.

• KSII Transactions on Internet and Information Systems (TIIS)
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• 제14권2호
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• pp.654-670
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• 2020

Workers at construction sites are easily exposed to many dangers and accidents involving falls, tripping, and missteps on stairs. However, researches on construction site monitoring system to prevent work-related injuries are still insufficient. The purpose of this study was to develop a wearable textile pressure insole sensor and examine its effectiveness in managing the real-time safety of construction workers. The sensor was designed based on the principles of parallel capacitance measurement using conductive textile and the monitoring system was developed by C# language. Three separate experiments were carried out for performance evaluation of the proposed sensor: (1) varying the distance between two capacitance plates to examine changes in capacitance charges, (2) repeatedly applying 1 N of pressure for 5,000 times to evaluate consistency, and (3) gradually increasing force by 1 N (from 1 N to 46 N) to test the linearity of the sensor value. Five subjects participated in our pilot test, which examined whether ascending and descending the stairs can be distinguished by our sensor and by weka assessment tool using k-NN algorithm. The 10-fold cross-validation method was used for analysis and the results of accuracy in identifying stair ascending and descending were 87.2% and 90.9%, respectively. By applying our sensor, the type of activity, weight-shifting patterns for balance control, and plantar pressure distribution for postural changes of the construction workers can be detected. The results of this study can be the basis for future sensor-based monitoring device development studies and fall prediction researches for construction workers.

• BMB Reports
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• 제30권1호
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• pp.46-54
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• 1997

Alkaliphilic Bacillus sp. S-1 secretes a large amount (approximately 80% of total pullulanase activity) of an extracellular pullulanase (PUL-E). The pullulanase exists in two forms: a precursor form (PUL-I: $M_r$ 180,000), and a processed form (PUL-E: $M_r$ 140,000). Two forms were purified to homogeneity and their properties were compared. PUL-I was different in molecular weight, isoelectric point, $NH_2$-terminal amino acid sequence, and stabilities over pH and temperature ranges. The catalytic activities of PUL-I were also distinguishable in the $K_m$ and $V_{max}$ values for various substrates, and in the specific activity for pullulan hydrolysis. PUL-E showed 10-fold higher specific activities than PUL-I. However. PUL-I is immunologically identical to PUL-E, suggesting that PUL-I is initially synthesized and proteolytically processed to the mature form of PUL-E. Processing was inhibited by PMSF, but not by pepstatin, suggesting that some intracellular serine proteases could be responsible for processing of the PUL-I. PUL-I has a different conformational structure for antibody recognition from that of PUL-E. It is also postulated that the translocation of alkaline pullulanase(AP) in the bacterium possibly requires processing of the $NH_2$-terminal region of the AP protein. Processing of the precursor involves a conformational shift. resulting in a mature form. Therefore. precursor processing not only cleaves the signal peptide, but also induces conformational shift. allowing development of active form of the enzyme.

• KSBB Journal
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• 제31권4호
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• pp.270-276
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• 2016

In glycoprotein, Terminal sialic acid residues of N-linked glycan are imperative things because they prevent the recognition from asialoglycoprotein-receptor that affect the half-life of glycoproteins. So establishment of culture process for enhancing sialic acid is important to maximize sialic acid contents of glycoprotein. In this study, we investigated effects of biphasic culture of Chinese hamster ovary (CHO) cells producing albumin-erythropoietin to increase sialylation. Biphasic cultures were performed with shift of $CO_2$ concentrations and temperatures at day 5 when viable cell density was decreased and sialidase was started to be released by cell lysis. The examined temperature set points were 33, 35 and $37^{\circ}C$ respectively and the $CO_2$ concentration was 1, 5, 10 and 15%. We confirmed that sialidase activity was the lowest in biphasic culture that was shifted from normal culture condition to 1% of $CO_2$ and $33^{\circ}C$ on day 5. However, the temperature and concentration of $CO_2$ have little effect on activity of ${\alpha}2,3$-sialyltransferase. Also, sialic acid contents were enhanced 1.13-fold higher than that in control culture. In conclusion, Biphasic cultivation in CHO cells led to inhibition of sialidase activity and increases of sialylated glycan.

• Biotechnology and Bioprocess Engineering:BBE
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• 제7권1호
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• pp.1-5
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• 2002

A fusion protein, consisting of a human epidermal growth factor (hEGF) as the recognition domain and human angiogenin as the toxin domain, can be used as a targeted therapeutic against breast cancer cells among others. The fusion protein was expressed as inclusion body in recombinant E. coli, and when the conventional, solution-phase refolding process was used the refolding yield was very low due to severe aggregation. It was probably because of the opposite electric charge at a neutral pH resulting from the vastly different pI values of each domain. The solid-phase refolding process that exploited the ionic interactions between ionic exchanger surface and the fusion protein was tried, but the adsorption yield was also very low, below $ 30\%$, regardless of the resins and pH conditions used. Therefore, to provide a higher ionic affinity toward the solid matrix, six lysine residues were tagged to the N-terminus of the hEGF domain. When heparin-Sepharose was used as the matrix, the adsorption capacity increased 2.5-3 times to about $88\%$. Besides the intrinsic affinity of angiogenin to heparin, the poly-lysine tag provided additional ionic affinity. And the subsequent refolding yield increased nearly 13-fold, from ca. $4.8\%$ in the conventional refolding of the untagged fusion protein to $63.6\%$. The process was highly reproducible. The refolded protein in the column eluate retained RNase bioactivity of angiogenin.

• Genomics & Informatics
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• 제12권4호
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• pp.283-288
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• 2014

Among all serious diseases globally, diabetes (type 1 and type 2) still poses a major challenge to the world population. Several target proteins have been identified, and the etiology causing diabetes has been reasonably well studied. But, there is still a gap in deciding on the choice of a drug, especially when the target is mutated. Mutations in the KCNJ11 gene, encoding the kir6.2 channel, are reported to be associated with congenital hyperinsulinism, having a major impact in causing type 1 diabetes, and due to the lack of its 3D structure, an attempt has been made to predict the structure of kir6.2, applying fold recognition methods. The current work is intended to investigate the affinity of four phytochemicals namely, curcumin (Curcuma longa), genistein (Genista tinctoria), piperine (Piper nigrum), and pterostilbene (Vitis vinifera) in a normal as well as in a mutant kir6.2 model by adopting a molecular docking methodology. The phytochemicals were docked in both wild and mutated kir6.2 models in two rounds: blind docking followed by ATP-binding pocket-specific docking. From the binding pockets, the common interacting amino acid residues participating strongly within the binding pocket were identified and compared. From the study, we conclude that these phytochemicals have strong affinity in both the normal and mutant kir6.2 model. This work would be helpful for further study of the phytochemicals above for the treatment of type 1 diabetes by targeting the kir6.2 channel.

• Molecules and Cells
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• 제45권8호
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• pp.575-587
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• 2022

Human ABCB6 is an ATP-binding cassette transporter that regulates heme biosynthesis by translocating various porphyrins from the cytoplasm into the mitochondria. Here we report the cryo-electron microscopy (cryo-EM) structures of human ABCB6 with its substrates, coproporphyrin III (CPIII) and hemin, at 3.5 and 3.7 Å resolution, respectively. Metal-free porphyrin CPIII binds to ABCB6 within the central cavity, where its propionic acids form hydrogen bonds with the highly conserved Y550. The resulting structure has an overall fold similar to the inward-facing apo structure, but the two nucleotide-binding domains (NBDs) are slightly closer to each other. In contrast, when ABCB6 binds a metal-centered porphyrin hemin in complex with two glutathione molecules (1 hemin: 2 glutathione), the two NBDs end up much closer together, aligning them to bind and hydrolyze ATP more efficiently. In our structures, a glycine-rich and highly flexible "bulge" loop on TM helix 7 undergoes significant conformational changes associated with substrate binding. Our findings suggest that ABCB6 utilizes at least two distinct mechanisms to fine-tune substrate specificity and transport efficiency.

• The Korean Journal of Ecology
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• 제24권3호
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• pp.191-202
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• 2001

인간은 생활환경을 확장하고 그 환경의 질을 향상시키기 위해 문명화를 진전시키는 과정에서 많은 자연환경을 양적으로 잠식하고, 그 질을 저하시켜 왔다. 이러한 인간활동을 많은 생물종을 사라지게 하였고, 인간자신도 그 자신의 존재를 위협하는 환경문제에 직면하게 하였다. 이러한 사실을 인식한 인간의 반응이 복원생태학의 성립과 발전으로 표출되고 있다. 복원생태학은 온전한 자연의 체계와 기능을 모방하여 훼손된 환경을 치유하는 환경기술이다. 이러한 복원은 훼손의 정도에 따라 처리방법이 다르고, 그 규모나 목표에 따라서도 그 종류를 구분할 수 있다. 따라서 그 시행을 위해서는 생태학적 진단이 필요하고, 그러한 진단결과에 기초하여 처방할 때 비용과 노력을 절약하고 상정한 목표에도 도달할 수 있다. 이런 점에서 복원생태학은 기존의 생태학에 토대를 두고 현실세계를 탐구하는 실용학문이며, 그에 더하여 기존의 생태학적 연구를 통하여 획득된 개념과 이론을 현장에서 검증하는 하나의 현장실습인 것이다. 인간은 지구라는 생태계에서 생활하면서 어떤 면에서 무리하게 자연을 이용하였지만, 어떤 면에서 불가피하게 자연을 훼손해 왔다. 더구나 이러한 이용과 훼손은 앞으로도 지속될 전망이다. 그러한 현상이 인류의 생존을 위협하는 환경문제를 몰고 오는 오늘의 시점에서 우리는 아무런 대비없이 그 생활을 계속 이어갈 수는 없다. 불필요한 이용을 줄이고, 부족한 부분을 보충하며, 취약한 부분을 보완하는 생태학적 복원을 추구하여 다가 올 미래의 환경위기에 대비하여야 한다. 이런 점에서 복원생태학은 미래를 위한 생태학으로 인정될 수 있을 것이다.