• 제목/요약/키워드: C33F10_2

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Cloning, Expression, Purification, and Properties of an Endoglucanase Gene (Glycosyl Hydrolase Family 12) from Aspergillus niger VTCC-F021 in Pichia pastoris

  • Pham, Thi Hoa;Quyen, Dinh Thi;Nghiem, Ngoc Minh;Vu, Thu Doan
    • Journal of Microbiology and Biotechnology
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    • 제21권10호
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    • pp.1012-1020
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    • 2011
  • A gene coding for an endoglucanase (EglA), of the glycosyl hydrolase family 12 and derived from Aspergillus niger VTCC-F021, was cloned and sequenced. The cDNA sequence, 717 bp, and its putative endoglucanase, a 238 aa protein with a predicted molecular mass of 26 kDa and a pI of 4.35, exhibited 98.3-98.7% and 98.3-98.6% identities, respectively, with cDNA sequences and their corresponding endoglucanases from Aspergillus niger strains from the GenBank. The cDNA was overexpressed in Pichia pastoris GS115 under the control of an AOX1 promoter with a level of 1.59 U/ml culture supernatant, after 72 h of growth in a YP medium induced with 1% (v/v) of methanol. The molecular mass of the purified EglA, determined by SDS-PAGE, was 33 kDa, with a specific activity of 100.16 and 19.91 U/mg toward 1% (w/v) of ${\beta}$-glucan and CMC, respectively. Optimal enzymatic activity was noted at a temperature of $55^{\circ}C$ and a pH of 5. The recombinant EglA (rEglA) was stable over a temperature range of $30-37^{\circ}C$ and at pH range of 3.5-4.5. Metal ions, detergents, and solvents tested indicated a slightly inhibitory effect on rEglA activity. Kinetic constants ($K_m$, $V_{max}$, $k_{cat}$, and $k_{cat}/K_m$) determined for rEglA with ${\beta}$-glucan as a substrate were 4.04 mg/ml, 102.04 U/mg, 2,040.82 $min^{-1}$, and 505.05, whereas they were 10.17 mg/ml, 28.99 U/mg, 571.71 $min^{-1}$, and 57.01 with CMC as a substrate, respectively. The results thus indicate that the rEglA obtained in this study is highly specific toward ${\beta}$-glucan. The biochemical properties of rEglA make it highly valuable for downstream biotechnological applications, including potential use as a feed enzyme.

Structure-Activity Relationships of Polyhydroxyursane-type Triterpenoids on the Cytoprotective and Anti-inflammatory Effects

  • Jung, Hyun-Ju;Nam, Jung-Hwan;Lee, Kyung-Tae;Lee, Yong-Sup;Choi, Jong-Won;Kim, Won-Bae;Chung, Won-Yoon;Park, Kwang-Kyun;Park, Hee-Juhn
    • Natural Product Sciences
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    • 제13권1호
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    • pp.33-39
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    • 2007
  • Eleven polyhydroxyursane triterpenoids (PHUTs) were tested to determine their cytoprotective, immunosuppressive and anti-inflammatory effects. To compare the bioactivities of $19{\alpha}$-hydroxyursane-type triterpenoids {23-hydroxytormentic acid (6), its methyl ester (7), tormentic acid (8), niga-ichigoside $F_1$ (9),euscaphic acid (10) and kaji-ichigoside $F_1$ (11)} of the Rosaceae crude drugs (Rubi Fructus and Rosa rugosae Radix) with PHUTs possessing no $19{\alpha}-hydroxyl$ of Centella asiatica (Umbelliferae), the four PHUTs, asiaticoside (1), madecassoside (2), asiatic acid (3), and madecassic acid (4) were isolated from C. asiatica and 23-hydroxyursolic acid (5) from Cussonia bancoensis. Cytoprotective effects were assessed by measuring cell viabilities against cisplatin-induced cytotoxocity in $LLC-PK_1$, cells (proximal tubule, pig kidney) to determine whether these agents have protective effects against nephrotoxicity caused by cisplatin. The inhibitory effect of 11 PHUTS on nitric oxide (NO) and prostaglandin $E_2\;(PGE_2)$ were evaluated by measuring nitrite accumulation in lipopolysaccharide (LPS)-induced macrophage RAW 264.7 cells, and their anti-inflammatory effects were tested in 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced mouse ear edema model. Six MHUTs (compounds 1, 2, 4, 6, 10, and 11) exhibited higher cell viabilities during cisplatin-induced cytotoxicity testing even at a concentration of $200\;{\mu}g/ml$ than cisplatin only-treated group, suggesting that ese compounds have the potentcytoprotective efffcts. Compounds 1 and 3 of the C. asiatica and niga-ichigoside $F_1$ exhibited no inhibitory effect on NO and/or $PGE_2$ production whereas other PHUTs produced mild to significant NO and/or $PGE_2$ production.The four compounds (2, 5, 9, and 10) potently inhibited mouse ear edema induced by TPA whereas two compounds (1 and 3) had no activity in this test. These results suggest that many PHUTs are potentchemopreventives. Structure-activity relationship (SAR) was also discussed in each assay with regard to the significant role of OHs at the position of 2, 3, 6, 19, and 23 and to the glycoside linkage at the 28-carboxyl.

발아 서목태 가수분해물의 근위축 억제 효과 (Germinated Rhynchosia nulubilis Hydrolysate Ameliorates Dexamethasone-induced Muscle Atrophy by Downregulating MAFbx Expression in C2C12 Cells and C57BL/6 Mice)

  • 이원경;김은지;김상곤;구영민;길영숙;신승미;안민주;강민철;하영술
    • 생명과학회지
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    • 제33권3호
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    • pp.277-286
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    • 2023
  • 스테로이드 유발 근위축은 장기간 코르티코스테로이드 요법의 일반적인 부작용이며 근육량과 기능의 현저한 감소로 이어질 수 있다. 발아된 Rhynchosia Nulubilis (GRH)는 항염증 및 항산화 특성이 있는 것으로 밝혀진 전통적인 약용 식물이다. 그러나 근 감소 억제 효능에 대하여는 보고된 바가 없다. 본 연구에서는 덱사메타손에 의해 유도된 근위축 C2C12 근관세포 모델과 마우스 모델에서 근위축 억제 활성에 대한 GRH의 영향을 평가하였다. 또한 골격근에 대한 GRH 효과의 기본이 되는 분자 경로를 확인하였다. May Grunwald-Giemsa 염색을 통한 근관 세포 염색 결과는 GRH 처리군에서 근관의 길이와 면적이 증가함을 보여주었다. 또한, muscle ring-finger protein-1 (MuRF-1)와 muscle atrophy F-box (MAFbx) 발현이 GRH 처리군에서 유의하게 감소하였다. 더욱이 덱사메타손에 의해 유도된 근위축 C57BL/6 마우스에서 트레드밀을 사용한 지구력 측정과 악력측정기를 이용한 악력 측정을 통한 근육 기능을 평가한 결과 GRH를 경구 투여한 마우스의 근육 기능이 향상되었다. 또한 이러한 결과는 근육 조직에서 GRH에 의한 MAFbx의 발현 억제를 통해 일어남을 확인하였다. 결론적으로 GRH는 MAFbx 발현을 억제하여 유비퀴틴-프로테아좀 경로를 차단함으로써 덱사메타손에 의해 유발된 근위축을 개선할 수 있을 것이다.

Identification of Coupling and Repulsion Phase DNA Marker Associated With an Allele of a Gene Conferring Host Plant Resistance to Pigeonpea sterility mosaic virus (PPSMV) in Pigeonpea (Cajanus cajan L. Millsp.)

  • Daspute, Abhijit;Fakrudin, B.
    • The Plant Pathology Journal
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    • 제31권1호
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    • pp.33-40
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    • 2015
  • Pigeonpea Sterility Mosaic Disease (PSMD) is an important foliar disease caused by Pigeonpea sterility mosaic virus (PPSMV) which is transmitted by eriophyid mites (Aceria cajani Channabasavanna). In present study, a F2 mapping population comprising 325 individuals was developed by crossing PSMD susceptible genotype (Gullyal white) and PSMD resistant genotype (BSMR 736). We identified a set of 32 out of 300 short decamer random DNA markers that showed polymorphism between Gullyal white and BSMR 736 parents. Among them, eleven DNA markers showed polymorphism including coupling and repulsion phase type of polymorphism across the parents. Bulked Segregant Analysis (BSA), revealed that the DNA marker, IABTPPN7, produced a single coupling phase marker (IABTPPN $7_{414}$) and a repulsion phase marker (IABTPPN $7_{983}$) co-segregating with PSMD reaction. Screening of 325 F2 population using IABTPPN7 revealed that the repulsion phase marker, IABTPPN $7_{983}$, was co-segregating with the PSMD responsive SV1 at a distance of 23.9 cM for Bidar PPSMV isolate. On the other hand, the coupling phase marker IABTPPN $7_{414}$ did not show any linkage with PSMD resistance. Additionally, single marker analysis both IABTPPN $7_{983}$ (P<0.0001) and IABTPPN $7_{414}$ (P<0.0001) recorded a significant association with the PSMD resistance and explained a phenotypic variance of 31 and 36% respectively in $F_2$ population. The repulsion phase marker, IABTPPN7983, could be of use in Marker-Assisted Selection (MAS) in the PPSMV resistance breeding programmes of pigeonpea.

유한요소 시뮬레이션을 통한 지역난방열배관 특성 평가 및 강화이형관의 제안 (Design Validation and Improvement of District Heating Pipe Using FE Simulation)

  • 김주용;김호범;고현일;안영모;조종두
    • 대한기계학회논문집A
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    • 제33권4호
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    • pp.337-345
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    • 2009
  • This paper investigates the reliability of district heating pipes at thermo-elastic fatigue loading. District heating pipes, subjected to $120^{\circ}C$ and $16kg_f/cm^2$ due to water distributing service through inside the pipes, should endure long term cyclic thermal-mechanical loadings. The heating pipes are the co-centric tubes of steel pipe, poly urethane(PUR) insulator, and high density poly ethylene(HDPE) case. On installation, foam pad is externally wrapped for accommodating stress reduction near the bend sections of pipes. However, there have been frequent reports on the failures of bend sections in the middle of long term service. This study scrutinizes the observed failures near the bend sections through applying the finite element methods. Specially in this study, heating pipes are studied on the influence of foam padding on failures and proposed new designs for reinforced bend without foam pad.

울산과 평택항에 정박된 국제상선의 평형수에서 소형식물플랑크톤의 활성능력 (Ballast-water Microphytoplankton Diversity and Survivability from International Ships Berthed at Ulsan and Pyeongtaek Ports, Korea)

  • 백승호;장민철;장풍국;신경순
    • Ocean and Polar Research
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    • 제33권2호
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    • pp.113-125
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    • 2011
  • In order to assess the survival success of microphytoplankton species in ship ballast water, we examined microphytoplankton diversity from international commercial ships berthed at Ulsan and Pyeongtaek Ports, Korea, and also subjected them to laboratory studies. The ages of ballast water in each ship ranged from 1 to 365 days. Vessels originated from coastal China (Weihai, Lianyunsang and Shanghai), Chile, and from the Yellow and Pacific Oceans. The numbers of species and phytoplankton standing crops in uploaded ballast water were significantly related to the age of ballast water. The most diverse taxonomic group was diatoms. In the laboratory study, the value of in vivo fluorescence in M/V Spring Lyra gradually increased with increasing nutrients such as nitrate and phosphate. Phytoplankton in new (9 days), medium (31 days) and old (365 days) ballast water successfully survived under typical nutrient condition of port water and F/2 medium at $15^{\circ}C$ and $20^{\circ}C$, whereas phytoplankton in ballast water treatment did not survive, regardless of optimal temperature. Colonization process was dominated by diatoms; Skeletonema coastatum for M/V Spring Lyra, Thalassiosira pseudonana and Thalassiosira for M/V Han Yang, Thalassiosira pacifica and Odontella aurita for M/V Modern Express, and Chaetoceros pseudocurvisetus and Pseudo-nitzschia seriata for M/V Asian Legend. The successful establishment of non-native species was also related to nutrient richness. Our laboratory design can be applied as a practical tool to assess the survivability of invasive microphytoplankton introduced into local waters of Ulsan and Pyeongtaek.

Trophic position and diet shift based on the body size of Coreoperca kawamebari (Temminck & Schlegel, 1843)

  • Choi, Jong-Yun;Kim, Seong-Ki;Kim, Jeong-Cheol;Yoon, Jung-Do
    • Journal of Ecology and Environment
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    • 제44권1호
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    • pp.1-7
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    • 2020
  • Background: Fish body size is a major determinant of freshwater trophic interactions, yet only a few studies have explored the relationship between the fish body size and trophic interactions in river upstream. In this study, we investigated the relationship between the body size and trophic position (TP) of Coreoperca kawamebari (Temminck & Schlegel, 1843) in an upstream of the Geum River. Results: A stable isotope analysis (based on δ15N) was used to determine the TP based on the body size of C. kawamebari. The regression analysis (n = 33, f = 63.840, r2 = 0.68) clearly showed the relationship between the body length and TP of C. kawamebari. The TP of C. kawamebari was clearly divided by body size into the following classes: individuals of size < 10 cm that feed on insects and individuals of size > 10 cm feed on juvenile fish. This selective feeding is an evolutionarily selective tendency to maximize energy intake per unit time. Furthermore, the diet shift of C. kawamebari was led by different spatial distributions. The littoral zone was occupied by individuals of size < 10 cm, and those of size > 10 cm were mainly in the central zone. The littoral zone can be assumed to be enriched with food items such as ephemeropterans and dipterans. Conclusion: The TP of C. kawamebari, as a carnivorous predator, will have a strong influence on biotic interactions in the upstream area of the Geum River, which can lead to food web implication.

수확전(收穫前) 논의 벼줄기에 감염(感染)된 Fusarium moniliforme에서의 Gibberella 자낭각(子囊殼)의 발생(發生) (Before Harvest Occurrence of Gibberella Perithecia of Fusarium moniliforme on Infected Rice Stems In field)

  • 성재모;스나이더
    • 한국균학회지
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    • 제5권1호
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    • pp.33-37
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    • 1977
  • 포장(圃場)에서 키다리병(病)에 걸린 줄기와 종자(種子)로 부터 얻은 자낭각이 어느 종류(種類)의 Gibberella에 존재(存在)하며, 수확후(收穫後)에 얻은 자낭각의 생존(生存)여부를 조사(調査)하기 위하여 처리한 결과(結果)는 다음과 같다. 1. 종자소독(種子消毒)을 하지 않은 포장(圃場)에서 키다리병(病)의 이병율(罹病率)은 평균(平均) 12%이었다. 2. 수확전(收穫前)에 얻은 자낭각은 키다리병의 자낭각이 적미병의 자낭각보다 많았으나 종자(種子)나 봄에 채집한 자낭각은 거의 적미병에 있었다. 3. 키다리병에 이병된 벼에서 채종한 종자(種子)에서 키다리병, 적미병, Ophiobolus sp.는 10, 25, 8%로 나타났으며 이병(罹病)된 줄기로부터 얻은 포자(胞子)로 접종(接種)한 종자(種子)에서는 3, 10, 2%로 나타났다. 4. 적미병자낭각을 야외 땅속 실내 집눌속에 놓고 5개월이 지난다음 생존(生存)여부를 조사(調査)한 결과(結果) 야외에 놓은 것만 발아하지 않았다.

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한반도 서부유입권역에서 대기 중 에어로졸 성분의 화학적 특성 연구 II. 입자의 산성도 및 산화 특성 (A Study on Chemical Characteristics of Aerosol Composition at West Inflow Regions in the Korean Peninsula II. Characteristics of Inorganic Aerosol Acidity and Organic Aerosol Oxidation)

  • 최진수;김정호;이태형;최용주;박태현;안준영;박진수;김현재;구윤서;김신도;홍유덕;홍지형
    • 한국대기환경학회지
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    • 제32권5호
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    • pp.485-500
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    • 2016
  • We examined acidity state of inorganic aerosol and oxidation state of organic aerosol by High Resolution Time of Flight Aerosol Mass Spectrometer (HR-ToF-AMS) at Baengnyeong Super site from Jan 2012 to Dec 2013. Additionally, we carried out the analysis for the aerosol component group of organic matter ($C_xH_y$, $C_xH_yO_1$, $C_xH_yO_z$, $C_xH_yO_zN_p$) and elemental composition to calculate H/C, O/C, N/C, OM/OC and identify the oxidation state. The aerosol chemical composition in this study is dominated by sulfate ($SO_4{^{2-}}$), nitrate ($NO_3{^-}$) plays a smaller role in aerosol acidity. Ammonium ($NH_4{^+}$) was found in a formation of $(NH_4)_3H(SO_4)_2$. However, the binding formations of $NH_4NO_3$ and $NH_4Cl$ increase in the winter. $C_xH_yO_1$ indicating the oxidized state of $PM_{1.0}$ has the highest ratio of 41% while $C_xH_y$ indicating the non-oxidized state has a lower ratio of 36%, meaning that the oxidation level of $PM_{1.0}$ in Baengnyeong Island is high. The ratio between H/C and O/C was 1.33 and 0.78 respectively, showing the characteristic of LV-OOA (Low volatility-Oxygenated Organic Aerosol). Acidic and oxidized aerosols sampled during this field study were largely anthropogenic in origin from Chinese continent and photochemically aged.

The Relationship between Milk Protein Phenotypes and Lactation Traits in Brown Swiss and Canadienne

  • Kim, S.;Ng-Kwai-Hang, K.F.;Hayes, J.F.
    • Asian-Australasian Journal of Animal Sciences
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    • 제11권3호
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    • pp.311-317
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    • 1998
  • A total of 1033 Brown Swiss and 610 Canadienne cows were phenotyped for the genetic variants ${\alpha}_{s1}$-casein, ${\beta}$-casein, ${\kappa}$-casein, ${\beta}$-lactoglobulin and ${\alpha}$-lactalbumin. In Brown Swiss, frequency distributions were: 97.3% B and 2.7% C variant of ${\alpha}_{s1}$-casein; 31.6% $A^1$, 51.8% $A^2$, 0.5% $A^3$ and 16.1% B variant of ${\beta}$-casein; 70.4% A, 29.3% B, and 0.3% C variant of ${\kappa}$-casein; 41.7% A and 58.3% B variant of ${\beta}$-lactoglobulin; and 100% B variant of ${\alpha}$-lactalbumin. Corresponding frequencies in Canadienne for those five milk proteins were: 98.6 and 1.4%;58.5, 33.5, 0.08 and 7.9%; 78.8, 21.1 and 0.1%, 42.4 and 57.6%; and 100%. Analysis of variance by least squares showed possible association between milk protein phenotypes and some lactational production traits. There were no significant association of phenotypes of ${\alpha}_{s1}$-casein, ${\beta}$-casein and ${\beta}$-lactoglobulin with milk yield, fat yield, protein yield, fat percentage and protein percentage in both breeds during the three lactations. In the Brown Swiss, ${\kappa}$-casein phenotype was associated with 305-day fat yield and protein yield during the first lactation. ${\kappa}$-Casein AB was associated with higher milk, fat and protein yield during the second lactation. During the third lactation, ${\beta}$-lactoglobulin AA in Canadienne cows was associated with higher protein content in the milk (3.70%) when compared to phenotypes AB (3.54%) and BB (3.64%).