• Title/Summary/Keyword: GS5

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Cloning, High-Level Expression, Purification, and Properties of a Novel Endo-${\beta}$-1,4-Mannanase from Bacillus subtilis G1 in Pichia pastoris

  • Vu, Thi Thu Hang;Quyen, Dinh Thi;Dao, Thi Tuyet;Nguyen, Sy Le Thanh
    • Journal of Microbiology and Biotechnology
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    • v.22 no.3
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    • pp.331-338
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    • 2012
  • A novel gene coding for an endo-${\beta}$-1,4-mannanase (manA) from Bacillus subtilis strain G1 was cloned and overexpressed in P. pastoris GS115, and the enzyme was purified and characterized. The manA gene consisted of an open reading frame of 1,092 nucleotides, encoding a 364-aa protein, with a predicted molecular mass of 41 kDa. The ${\beta}$-mannanase showed an identity of 90.2-92.9% ${\leq}95%$) with the corresponding amino acid sequences from B. subtilis strains deposited in GenBank. The purified ${\beta}$-mannanase was a monomeric protein on SDS-PAGE with a specific activity of 2,718 U/mg and identified by MALDI-TOF mass spectrometry. The recombinant ${\beta}$-mannanase had an optimum temperature of $45^{\circ}C$ and optimum pH of 6.5. The enzyme was stable at temperatures up to $50^{\circ}C$ (for 8 h) and in the pH range of 5-9. EDTA and most tested metal ions showed a slightly to an obviously inhibitory effect on enzyme activity, whereas metal ions ($Hg^{2+}$, $Pb^{2+}$, and $Co^{2+}$) substantially inhibited the recombinant ${\beta}$-mannanase. The chemical additives including detergents (Triton X-100, Tween 20, and SDS) and organic solvents (methanol, ethanol, n-butanol, and acetone) decreased the enzyme activity, and especially no enzyme activity was observed by addition of SDS at the concentrations of 0.25-1.0% (w/v) or n-butanol at the concentrations of 20-30% (v/v). These results suggested that the ${\beta}$-mannanase expressed in P. pastoris could potentially be used as an additive in the feed for monogastric animals.

Efficient Expression, Purification, and Characterization of a Novel FAD-Dependent Glucose Dehydrogenase from Aspergillus terreus in Pichia pastoris

  • Yang, Yufeng;Huang, Lei;Wang, Jufang;Wang, Xiaoning;Xu, Zhinan
    • Journal of Microbiology and Biotechnology
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    • v.24 no.11
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    • pp.1516-1524
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    • 2014
  • Flavin adenine dinucleotide-dependent glucose dehydrogenase (FAD-GDH) can utilize a variety of external electron acceptors and also has stricter substrate specificity than any other glucose oxidoreductases, which makes it the ideal diagnostic enzyme in the field of glucose biosensors. A gene coding for a hypothetical protein, similar to glucose oxidase and derived from Aspergillus terreus NIH2624, was overexpressed in Pichia pastoris GS115 under the control of an AOX1 promoter with a level of 260,000 U/l in the culture supernatant after fed-batch cultivation for 84 h. After a three-step purification protocol that included isopropanol precipitation, affinity chromatography, and a second isopropanol precipitation, recombinant FAD-GDH was purified with a recovery of 65%. This is the first time that isopropanol precipitation has been used to concentrate a fermentation supernatant and exchange buffers after affinity chromatography purification. The purified FAD-GDH exhibited a broad and diffuse band between 83 and 150 kDa. The recombinant FAD-GDH was stable across a wide pH range (3.5 to 9.0) with maximum activity at pH 7.5 and $55^{\circ}C$. In addition, it displayed very high thermal stability, with a half-life of 82 min at $60^{\circ}C$. These characteristics indicate that FAD-GDH will be useful in the field of glucose biosensors.

Ginsenoside Rh2 epigenetically regulates cell-mediated immune pathway to inhibit proliferation of MCF-7 breast cancer cells

  • Lee, Hyunkyung;Lee, Seungyeon;Jeong, Dawoon;Kim, Sun Jung
    • Journal of Ginseng Research
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    • v.42 no.4
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    • pp.455-462
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    • 2018
  • Background: Ginsenoside Rh2 has been known to enhance the activity of immune cells, as well as to inhibit the growth of tumor cells. Although the repertoire of genes regulated by Rh2 is well-known in many cancer cells, the epigenetic regulation has yet to be determined, especially for comprehensive approaches to detect methylation changes. Methods: The effect of Rh2 on genome-wide DNA methylation changes in breast cancer cells was examined by treating cultured MCF-7 with Rh2. Pyrosequencing analysis was carried out to measure the methylation level of a global methylation marker, LINE1. Genome-wide methylation analysis was carried out to identify epigenetically regulated genes and to elucidate the most prominent signaling pathway affected by Rh2. Apoptosis and proliferation were monitored to examine the cellular effect of Rh2. Results: LINE1 showed induction of hypomethylation at specific CpGs by 1.6-9.1% (p < 0.05). Genome-wide methylation analysis identified the "cell-mediated immune response"-related pathway as the top network. Cell proliferation of MCF-7 was retarded by Rh2 in a dose-dependent manner. Hypermethylated genes such as CASP1, INSL5, and OR52A1 showed downregulation in the Rh2-treated MCF-7, while hypomethylated genes such as CLINT1, ST3GAL4, and C1orf198 showed upregulation. Notably, a higher survival rate was associated with lower expression of INSL5 and OR52A1 in breast cancer patients, while with higher expression of CLINT1. Conclusion: The results indicate that Rh2 induces epigenetic methylation changes in genes involved in immune response and tumorigenesis, thereby contributing to enhanced immunogenicity and inhibiting the growth of cancer cells.

Damage Effects Modeling by Chlorine Leaks of Chemical Plants (화학공장의 염소 누출에 의한 피해 영향 모델링)

  • Jeong, Gyeong-Sam;Baik, Eun-Sun
    • Fire Science and Engineering
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    • v.32 no.3
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    • pp.76-87
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    • 2018
  • This study describes the damage effects modeling for a quantitative prediction about the hazardous distances from pressurized chlorine saturated liquid tank, which has two-phase leakage. The heavy gas, chlorine is an accidental substance that is used as a raw material and intermediate in chemical plants. Based on the evaluation method for damage prediction and accident effects assessment models, the operating conditions were set as the standard conditions to reveal the optimal variables on an accident due to the leakage of a liquid chlorine storage vessel. A model of the atmospheric diffusion model, ALOHA (V5.4.4) developed by USEPA and NOAA, which is used for a risk assessment of Off-site Risk Assessment (ORA), was used. The Yeosu National Industrial Complex is designated as a model site, which manufactures and handles large quantities of chemical substances. Weather-related variables and process variables for each scenario need to be modelled to derive the characteristics of leakage accidents. The estimated levels of concern (LOC) were calculated based on the Gaussian diffusion model. As a result of ALOHA modeling, the hazardous distance due to chlorine diffusion increased with increasing air temperature and the wind speed decreased and the atmospheric stability was stabilized.

A Numerical Investigation for Prediction of Shock Deceleration of Conical Impactor in Gas-Gun Tests (가스건 시험에서 원추형 충격자의 충격 감가속도 예측에 관한 전산해석적 연구)

  • Yoon, Hee;Oh, Jong Soo;Jung, Myung-Suk
    • Journal of the Computational Structural Engineering Institute of Korea
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    • v.32 no.5
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    • pp.279-286
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    • 2019
  • In this study, a numerical investigation is conducted for the shock deceleration prediction of a conical impactor in gas-gun tests. With the development of weapon systems, gas-gun tests are required to validate the survivability and structural reliability of devices under test (DUT) in high-G shock environments, such as those over ten thousand Gs or more. As shock endurance is highly influenced by various bird parameters, such as mass, velocity, and pressure, it is important to determine the appropriate test conditions to generate a high-G shock environment. However, experimental repetitive studies are inefficient to validate test conditions in terms of economic aspects. Therefore, a numerical technique is required to replace experimental gas-gun tests. Here, a numerical investigation is conducted with ANSYS AUTODYN using explicit code. Through this investigation, the dynamic behavior of DUT is presented. In addition, the results of numerical studies are verified through a comparison with the experimental results of a gas-gun test.

Characterization of simple sequence repeats (SSRs) in Pleurotus pulmonarius cultivars (산느타리(Pleurotus pulmonarius) 품종의 초위성체(simple sequence repeats) 특성구명)

  • Choi, Jong In;Na, Kyeong Sook;Oh, Min-Ji;Ryu, Jae-San
    • Journal of Mushroom
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    • v.19 no.4
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    • pp.341-346
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    • 2021
  • Simple sequence repeats (SSRs) were isolated from major Pleurotus pulmonarius cultivars in Korea, namely 'HS47' (monokaryon, gamete of 'Santari'), 'GB19' (monokaryon, gamete of 'Santari'), 'Hosan,' 'Yeoleumneutali1,' 'Sambok,' 'Gangsan,' 'Yaksan,' 'Jasan,' 'Hyangsan,' and 'Yeoleumneutali2,' and characterized via HiSeq genome sequencing and bioinformatic analysis. The genome sizes of the monokaryons 'HS47' and 'GB19' were estimated to be 37.3 and 37.2 Mb, respectively, and those of the other dikaryotic cultivars ranged from 47.1 to 61.1 Mb. A total of 711 (smallest) and 1,106 (1.5 times the smallest) SSRs were found in the 'HS47' and 'Gangsan' genomes, respectively. Hexanucleotide and octanucleotide motifs accounted for the top two fractions of all SSRs. CGA/TCG, A/T, and CTC/GAG were the most frequently detected nucleotides in the SSRs. Most of the SSRs were 21~30 nucleotides long (hypervariable for application), accounting for 70% of all lengths of SSRs.

Observation of reinforcing fibers in concrete upon bending failure by X-ray computed tomographic imaging

  • Seok Yong Lim;Kwang Soo Youm;Kwang Yeom Kim;Yong-Hoon Byun;Young K. Ju;Tae Sup Yun
    • Computers and Concrete
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    • v.31 no.5
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    • pp.433-442
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    • 2023
  • This study presents the visually observed behavior of fibers embedded in concrete samples that were subjected to a flexural bending test. Three types of fibers such as macro polypropylene, macro polyethylene, and the hybrid of steel and polyvinyl alcohol were mixed with cement by a designated mix ratio to prepare a total of nine specimens of each. The bending test was conducted by following ASTM C1609 with a net deflection of 2, 4, and 7 mm. The X-ray computed tomography (XCT) was carried out for 7 mm-deflection specimens. The original XCT images were post-processed to denoise the beam-hardening effect. Then, fiber, crack, and void were semi-manually segmented. The hybrid specimen showed the highest toughness compared to the other two types. Debonding based on 2D XCT sliced images was commonly observed for all three groups. The cement matrix near the crack surface often involved partially localized breakage in conjunction with debonding. The pullout was predominant for steel fibers that were partially slipped toward the crack. Crack bridging and rupture were not found presumably due to the image resolution and the level of energy dissipation for poly-fibers, while the XCT imaging was advantageous in evaluating the distribution and behavior of various fibers upon bending for fiber-reinforced concrete beam elements.

A Wheat Variety, "Hwanggeumal" with Good Bread Quality, Red Grain, Partial Waxy, Tolerance to PHS

  • Chon-Sik Kang;Chang-Hyun Choi;Kyeong-Hoon Kim;Kyeong-Min Kim;Go Eun Lee;Jin-Hee Park;Jong-min Ko
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2022.10a
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    • pp.203-203
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    • 2022
  • A new winter wheat(Triticum aestivum L.) cultivar "Hwanggeumal" was developed by the NICS(National Institute of Crop Science), RDA(Rular Dvelopment Administraion) in 2019. It was derived from a cross of the "Jokyoung//Kauz/Rayon" and "Jopoom" in 2008. It had advanced generation through bulk and pedigree method for seven years and designated line name "Jeonju398" after AYT(Advance Yield Trial) test for two years. And "Hwangeumal" was designated variety name after RYT(Regional Yield Trial) test in eight locations around Korea for two years from 2018 to 2019. Its heading date was April 19 and maturity date was May 31, which were similar to Jokyoung. "Hwanggeumal" had shorter plant height(75 cm) and spike length(7.1 cm), spikes per m2(699) and lower 1,000 grain weight(44.2 g) than "Jokyoung"(78 cm, 8.2 cm, 776, 46.6 g, respectively). "Hwanggeumal" was showed weak to winter hardiness and susceptible to powdery mildew but tolerance to PHS(Pre-harvest sprouting). The average grain yield in the AYT was 6.2 ton/ha, which were 10% more than "Jokyoung" And in the RYT was 5.1 ton/ha in upland and 4.4 ton/ha in paddy field, which were lower than "Jokyoung", respectively. "Hwanggeumal"s flour yield (71.4%) and flour lightness (91.82) showed similar to "Jokyung" and higher protein content (14.0%) and gluten content (10.3%) and SDS-sedimentation volume (60.3ml). These result showed that the "Hwanggeumal" dough strength of flour is strong than "Jokyung". "Hwanggeumal"s HMW-GS(High molecular weight gluten subunits) composition are Glu-D1 (5+10), Granule-bound starch synthase(GBSS) composition are Wx-A1 (a), Wx-B1 (b), Wx-D1 (a) and composition of Puroindolines are Pina-D1(a), Pinb-D1(b).

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An enhancement of GloSea5 ensemble weather forecast based on ANFIS (ANFIS를 활용한 GloSea5 앙상블 기상전망기법 개선)

  • Moon, Geon-Ho;Kim, Seon-Ho;Bae, Deg-Hyo
    • Journal of Korea Water Resources Association
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    • v.51 no.11
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    • pp.1031-1041
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    • 2018
  • ANFIS-based methodology for improving GloSea5 ensemble weather forecast is developed and evaluated in this study. The proposed method consists of two steps: pre & post processing. For ensemble prediction of GloSea5, weights are assigned to the ensemble members based on Optimal Weighting Method (OWM) in the pre-processing. Then, the bias of the results of pre-processed is corrected based on Model Output Statistics (MOS) method in the post-processing. The watershed of the Chungju multi-purpose dam in South Korea is selected as a study area. The results of evaluation indicated that the pre-processing step (CASE1), the post-processing step (CASE2), pre & post processing step (CASE3) results were significantly improved than the original GloSea5 bias correction (BC_GS5). Correction performance is better the order of CASE3, CASE1, CASE2. Also, the accuracy of pre-processing was improved during the season with high variability of precipitation. The post-processing step reduced the error that could not be smoothed by pre-processing step. It could be concluded that this methodology improved the ability of GloSea5 ensemble weather forecast by using ANFIS, especially, for the summer season with high variability of precipitation when applied both pre- and post-processing steps.

Identifying sources of heavy metal contamination in stream sediments using machine learning classifiers (기계학습 분류모델을 이용한 하천퇴적물의 중금속 오염원 식별)

  • Min Jeong Ban;Sangwook Shin;Dong Hoon Lee;Jeong-Gyu Kim;Hosik Lee;Young Kim;Jeong-Hun Park;ShunHwa Lee;Seon-Young Kim;Joo-Hyon Kang
    • Journal of Wetlands Research
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    • v.25 no.4
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    • pp.306-314
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    • 2023
  • Stream sediments are an important component of water quality management because they are receptors of various pollutants such as heavy metals and organic matters emitted from upland sources and can be secondary pollution sources, adversely affecting water environment. To effectively manage the stream sediments, identification of primary sources of sediment contamination and source-associated control strategies will be required. We evaluated the performance of machine learning models in identifying primary sources of sediment contamination based on the physico-chemical properties of stream sediments. A total of 356 stream sediment data sets of 18 quality parameters including 10 heavy metal species(Cd, Cu, Pb, Ni, As, Zn, Cr, Hg, Li, and Al), 3 soil parameters(clay, silt, and sand fractions), and 5 water quality parameters(water content, loss on ignition, total organic carbon, total nitrogen, and total phosphorous) were collected near abandoned metal mines and industrial complexes across the four major river basins in Korea. Two machine learning algorithms, linear discriminant analysis (LDA) and support vector machine (SVM) classifiers were used to classify the sediments into four cases of different combinations of the sampling period and locations (i.e., mine in dry season, mine in wet season, industrial complex in dry season, and industrial complex in wet season). Both models showed good performance in the classification, with SVM outperformed LDA; the accuracy values of LDA and SVM were 79.5% and 88.1%, respectively. An SVM ensemble model was used for multi-label classification of the multiple contamination sources inlcuding landuses in the upland areas within 1 km radius from the sampling sites. The results showed that the multi-label classifier was comparable performance with sinlgle-label SVM in classifying mines and industrial complexes, but was less accurate in classifying dominant land uses (50~60%). The poor performance of the multi-label SVM is likely due to the overfitting caused by small data sets compared to the complexity of the model. A larger data set might increase the performance of the machine learning models in identifying contamination sources.