• Title/Summary/Keyword: 43 kD protein

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Cloning, Expression, and Characterization of Bacillus sp. snu-7 Inulin Fructotransferase

  • Kim, Chung-Sei;Hong, Chang-Ki;Kim, Kyoung-Yun;Wang, Xiu-Ling;Kang, Su-Il;Kim, Su-Il
    • Journal of Microbiology and Biotechnology
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    • v.17 no.1
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    • pp.37-43
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    • 2007
  • A gene encoding inulin fructotransferase (di-D-fructofuranose 1,2': 2,3' dianhydride [DFA III]-producing IFTase, EC 4.2.2.18) from Bacillus sp. snu-7 was cloned. This gene was composed of a single, 1,353-bp open reading frame encoding a protein composed of a 40-amino acid signal peptide and a 410-amino acid mature protein. The deduced amino acid sequence was 98% identical to Arthrobacter globiformis C11-1 IFTase (DFA III-producing). The enzyme was successfully expressed in E. coli as a functionally active, His-tagged protein, and it was purified in a single step using immobilized metal affinity chromatography. The purified enzyme showed much higher specific activity (1,276 units/mg protein) than other DFA III-producing IFTases. The recombinant and native enzymes were optimally active in very similar pH and temperature conditions. With a 103-min half-life at $60^{\circ}C$, the recombinant enzyme was as stable as the native enzyme. Acidic residues and cysteines potentially involved in the catalytic mechanism are proposed based on an alignment with other IFTases and a DFA IIIase.

Effects of Crystalline Lysine, Threonine and Tryptophan Supplementation of Diets Containing Reduced Protein Levels on Performance of Growing Pigs

  • Li, D.F.;Xiao, C.T.;Kim, J.H.;Cho, W.T.;Han, In K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.11 no.1
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    • pp.43-50
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    • 1998
  • Three hundred and fifty-two growing pigs were utilized in three growth trials to investigate the effects of crystalline amino acid supplementation on growing pigs' growth performance and feed consumption. In Exp. 1, diets were formulated on the basis of digestible amino acids. Utilization of rapeseed meal plus cottonseed meal (R + C) in diet supported similar average daily gain (ADG), feed/gain (F/G) and cost per gain (p > 0.05) to group of soybean meal (SBM). In Exp. 2, 14% CP diets containing equal amount of cottonseed meal and rapeseed meal were formulated with the addition of threonine and/or tryptophan. Supplementation with threonine improved ADG and F/G (p < 0.05). Supplementation with only tryptophan made no benefits for ADG of growing pigs. In Exp. 3, diets with different threonine level were formulated. Increasing dietary threonine to 0.54% improved (p < 0.05) ADG and F/G compared to 0.45% dietary threonine. Increase in dietary threonine did not improve ADG (p > 0.05) but decreased F/G (p < 0.05) because of a decrease in average daily feed intake. Pigs fed diet with dietary threonine/lysine ratio of 67.5% supported optimum growth performance.

Purification of Glucose Oxidase by Affinity Chromatography and Its Characterization (친화성 크로마토그래피를 이용한 글루코오스 옥시다아제의 정제와 효소특성)

  • Ko Jung Hwan;Byun Si Myung
    • Journal of the Korean Chemical Society
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    • v.23 no.3
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    • pp.165-174
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    • 1979
  • A purification technique of glucose oxidase was developed. Using the gluconyl-${\omega}$-aminohexyl Sepharose affinity chromatography, it was partially purified 14.6 folds with 79.7% yield. With the combination of the affinity chromatography and Sepharose 6B gel filtration, the enzyme was purified 27.2 folds from the broth with 74.1% yield. The final purified preparation showed 90.83 U of glucose oxidase activity per mg of protein and a single band by 7% polyacrylamide gel electrophoresis. The absorption spectrum and substrate specificity of the enzyme were studied and the fianal preparation showed the optimal pH between 5.6 and 6.0, the optimal temperature at $40^{\circ}C$, $8.5{\times}10^{-3}M$ of $K_m$ for D-glucose, and 3.43 kcal/mole of the activation energy.

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Diagnostic value of C-reactive Protein and Vascular Endothelial Growth Factor in Differentiation of Pleural Effusions (흉막액 감별에 있어서 C-반응성단백과 혈관내피성장인자의 유용성)

  • Kim, Sang Ha;Lee, Won Yeon;Park, Joo Young;Park, Hyun Sook;Han, Hye-Kyoung;Ju, Hun Su;Hong, Tae Won;Lee, Nak Won;Shin, Kye Chul;Yong, Suk Joong
    • Tuberculosis and Respiratory Diseases
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    • v.55 no.5
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    • pp.467-477
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    • 2003
  • Background : Pleural effusions are generally divided into transudates and exudates. If it is exudative, more diagnostic tests are required in order to determine the cause of the local disease. A malignancy is a common and important cause of exudative pleural effusions. Because the pleural fluid cytology and pleural biopsy specimens do not provide a diagnosis in a high percentage of malignant effusions, several tumor markers have been examined. In order to overcome this limitation, this study hypothesized that C-reactive protein(CRP) and vascular endothelial growth factor(VEGF) measurements would be useful for differentiating trasudates from exudates and determining the differences between a benign and malignant effusion. Methods : Eighty consecutive patients with a pleural effusion (tuberculous 20, parapneumonic 20, malignant 20, transudative 20) were examined prospectively: 60 of them were classified according to Light's criteria as having an exudative fluid and 20 had a transudative fluid. The standard parameters of a pleural effusion were examined and the serum and pleural effusion VEGF levels were measured using enzyme linked immunosorbent assay(ELISA). CRP in the serum and pleural fluid was determined by a turbidimetric immunoassay. Results : The pleural CRP levels in the exudates were significantly higher than those in the transudates, $4.19{\pm}4.22mg/d{\ell}$ and $1.29{\pm}1.45mg/d{\ell}$, respectively. The VEGF levels in the pleural effusions were significantly elevated in the exudates compared to the transudate, $1,011{\pm}1,055pg/m{\ell}$ and $389{\pm}325pg/m{\ell}$, respectively. The VEGF ratio in the exudative effusion is significantly higher than a transudative effusions, $3.9{\pm}4.7$ and $1.6{\pm}0.9$, respectively. The pleural CRP levels in the patients with a benign effusion($4.15{\pm}4.20mg/d{\ell}$) were significantly higher than those in the malignant effusion($1.43{\pm}1.91mg/d{\ell}$). The VEGF ratio is significantly higher in malignant effusions($4.9{\pm}5.5$) than in benign effusions($2.8{\pm}3.6$). Conclusion : In conclusion, the CRP and VEGF levels in the serum and pleural effusion can distinguish between transudates and exudates. Moreover it can differentiate between benign and malignant pleural effusions.

Binding Studies of Cardiovascular Drug on ${\beta}$ Adrenoceptors in Rat Left Ventricle using $(-)-[^3H]-DHA$, $Non-{\beta}_1/{\beta}_2-selective$ Radioligand (${\beta}_1/{\beta}_2$ 비선택적 Radioligand $(-)-[^3H]-DHA$를 사용한 Rat 좌심실 ${\beta}-adrenoceptor$에 대한 심장순환계 약물의 Binding)

  • Kwon, Kwang-Il;Lee, Sun-Kyung;Yoo, Sung-Eun
    • The Korean Journal of Pharmacology
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    • v.27 no.2
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    • pp.119-123
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    • 1991
  • ${\beta}-Adrenoceptor$ binding study of ${\beta}-agonist$ ((-)NE), ${\beta}-antagonists$ $(({\pm})\;propranolol,\;labetalol)$ and PDE inhibitors (imazodan, KR-30045, KR-30075 etc.) was performed using $(-)-[^3H]-DHA$, as a $non-{\beta}_1/{\beta}_2$ selective radioligand. In saturation studies, $K_d$ and $B_{max}$ of $(-)-[^3H]-DHA$ to ${\beta}-adrenoceptors$ in rat left ventricle in which both ${\beta}_1$ and ${\beta}_2$ receptors coexist were determined to be $1.5{\pm}0.43\;nM$ and $22.0{\pm}0.9\;fmol/mg$ protein, respectively. $({\pm})Propranolol$, labetalol and (-)NE competed for $(-)-[^3H]-DHA$ binding sites in an essentialy monophasic manner with $Ki=17.0{\pm}0.40\;nM,\;57.3{\pm}1.30\;nM,\;and\;1.57{\pm}0.95\;{\mu}M$, respectively. All of PDE inhibitors inhibited the $(-)-[^3H]-DHA$ binding by only below 10% even at the high concentration of $10^{-3}M$. The present results suggest that propranolol, labetalol and NE are $non-{\beta}_1/{\beta}_2$ selective antagonists and agonist, respectively. Additionally, this study shows that imazodan and new synthesized PDE inhibitors may hardly have the affinities to ${\beta}-adrenoceptors$ in cardiac muscle.

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Hematologic and biochemical reference intervals for natural monument Korean goral (Naemorhedus caudatus) (천연기념물 한국 산양의 혈액 및 혈액 화학적 정상표준범위)

  • Ahn, Sangjin;Choi, Youngjin;Kim, Jong-Taek
    • Korean Journal of Veterinary Service
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    • v.40 no.2
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    • pp.155-159
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    • 2017
  • Korean goral (Neamorhedus caudatus) is registered as a natural monument number 217 by South Korea Cultural Heritage Administration. It is also recognized as the endangered species I by Convention on International Trade in Endangered Species of Wild Fauna and Flora (CITES). In this study, blood samples of Korean goral were collected to make reference intervals. Blood sampling was conducted on 19 numbers of Korean gorals (ten females, nine males), which were reared in Korean Goral Restoration Center. For total samples, the reference intervals of hematological parameters were: white blood cell $7.69{\sim}10.87K/{\mu}{\Lambda}$; hematocrit 36.73~46.18%; red blood cell $10.72{\sim}12.86K/{\mu}{\Lambda}$; hemoglobin 12.79~15.14 g/dL; mean corpuscular volume 33.15~36.75 fL; mean corpuscular hemoglobin 11.53~12.23 pg; mean corpuscular hemoglobin concentration 32.64-35.91 g/dL and red blood cell distribution width 39.2~43.46%. For total samples of each parameters, the following results were obtained for serum biochemistry: glucose 111.81~153.77 mg/dL; blood urea nitrogen 22.35~28.91 mg/dL; creatine 1.22~1.84 mg/dL; phosphate 4.57~6 mg/dL; calcium 8.7~9.1 mg/dL; total protein 6.53~6.92 g/dL; albumin 3.1~3.48 g/dL; globulin 3.26~3.62 g/dL; alanine aminotransferase 56.7~158.56 U/L; aspartate aminotransferase 230.35~473.06 U/L; alkaline phosphatase 178.06~332.47 U/L; gamma-glutamyl transpeptidase 131.6-~181.24 U/L; total bilirubin 1.47~2.12 mg/dL; cholesterol 46.48~71.52 mg/dL; amylase 16.3~26.03 U/L; sodium 150.43~153.88 mmol/L; potassium 3.98~4.6 mmol/L and chlorine 109.48~113.26 mmol/L. The ranges of values were similar campared to previous studies except in the case of RDW value, which showed higher range than the RDW value of a previous study. The reference intervals from this study will be useful data for treatment and management of gorals.

Inhibition of the expression on MMP-2, 9 and morphological changes via human fibrosarcoma cell line by 6,6'-bieckol from marine alga Ecklonia cava

  • Zhang, Chen;Li, Yong;Shi, Xiujuan;Kim, Se-Kwon
    • BMB Reports
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    • v.43 no.1
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    • pp.62-68
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    • 2010
  • Matrix Metalloproteinases (MMPs) are a family of zinc-endopeptidases which can degrade extracellular matrix (ECM) components and play important roles in a variety of biological and pathological processes. 6,6'-bieckol isolated and characterized from an edible marine brown alga Ecklonia cava (EC), according to the comprehensive spectral analysis of MS and NMR data. Here the influence of 6,6'-bieckol on expressions of MMPs was examined by zymography and western blot analysis via human fibrosarcoma cell line (HT1080). It is shown that 6,6'-bieckol significantly down regulated the expressions of MMP-2 and -9 in dose-dependent manner. The influence of 6,6'-bieckol on the cell viability and cell behavior of HT1080 cells were also investigated, our dates shown that it suppressed the migration and 3D culture in HT1080 cells. Meanwhile, we explored several signal pathways which may contribute to this process, and found the suppressing of MMPs expressions in HT1080 cells might be due to the suppression of NF-${\kappa}B$ signal pathway.

Variations in Reserved Nutrient Consumption and Growth of Pacific Oyster (Crassostra gigas) Larvae during Starvation (참굴 (Crassostrea gigas) 유생의 절식에 따른 성장 및 체내 에너지원의 소비변화)

  • Hur, Young-Baek;Kim, Tae-Eic;Lee, Seung-Ju;Hur, Sung-Bum
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.43 no.5
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    • pp.489-494
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    • 2010
  • The nutritional demand of oyster larva (Crassostrea gigas) were investigated to determine the optimal culture conditions and improve micro-algae utilization. Changes in nutrients and shell growth were examined in fed and 96-h (48 h in late umbone stage) oysters at four larval stages. Shell growth increased significantly in D shape larvae, regardless of feeding variations. No growth was observed in starved larvae, except in shell length of umbone (to 11.9 ${\mu}m$). Fed larvae showed significant growth in all development stages (P < 0.05). During starvation, lipids were most significantly decreased in all larval stages (by 76.8%, 68.3%, 76.3%, and 40.3%, respectively), followed by protein (41.1%, 31.1%, 33.1%, 16.7%) and nitrogen-free extracts (40.8%, 24.3%, 36.9%, 20.1%), Gross energy (kcal/g) consumption in each larval stage was 49.6%, 35.1%, 39.1%, and 20.4%, respectively. Our results indicate that lipids are the most important energy source during the early larval development stages of C. gigas.

Hydrogen Cyanide Produced by Pseudomonas chlororaphis O6 Exhibits Nematicidal Activity against Meloidogyne hapla

  • Kang, Beom Ryong;Anderson, Anne J.;Kim, Young Cheol
    • The Plant Pathology Journal
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    • v.34 no.1
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    • pp.35-43
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    • 2018
  • Root-knot nematodes (Meloidogyne spp.) are parasites that attack many field crops and orchard trees, and affect both the quantity and quality of the products. A root-colonizing bacterium, Pseudomonas chlororaphis O6, possesses beneficial traits including strong nematicidal activity. To determine the molecular mechanisms involved in the nematicidal activity of P. chlororaphis O6, we constructed two mutants; one lacking hydrogen cyanide production, and a second lacking an insecticidal toxin, FitD. Root drenching with wild-type P. chlororaphis O6 cells caused juvenile mortality in vitro and in planta. Efficacy was not altered in the fitD mutant compared to the wild-type but was reduced in both bioassays for the mutant lacking hydrogen cyanide production. The reduced number of galls on tomato plants caused by the wild-type strain was comparable to that of a standard chemical nematicide. These findings suggest that hydrogen cyanide-producing root colonizers, such as P. chlororaphis O6, could be formulated as "green" nematicides that are compatible with many crops and offer agricultural sustainability.

Proteomic analysis of serum proteins responsive to styrene exposure (Styrene 노출에 반응을 보이는 혈청 단백질에 대한 프로테오믹스 분석)

  • Kim, Ki-Woong;Heo, Kyung-Hwa;Won, Yong Lim;Jeong, Jin Wook;Kim, Tae Gyun;Park, Injeong
    • Journal of Korean Society of Occupational and Environmental Hygiene
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    • v.17 no.3
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    • pp.235-244
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    • 2007
  • By comparing the proteins from the workers exposed to styrene with the ones from controls, it may be possible to identify proteins that play a role in the occurrence and progress of occupational disease and thus to study the molecular mechanisms of occupational disease. In order to find the biomarkers for assessing the styrene effects early, before clinical symptoms develop and to understand the mechanisms of adverse health effects, we surveyed 134 employees, among whom 52 workers(30 male and 22 female) were chronically exposed to styrene in 10 glass-reinforced plastic boat manufacturing factories in Korea and 82 controls had never been occupationally exposed to hazardous chemicals including styrene. The age and drinking habits and serum biochemistry such as total protein, BUN and serum creatinine in both groups were significantly different. Exposed workers were divided into three groups according to exposure levels of styrene(G1, below 1/2 TLV; G2, 1/2 TLV to TLV; G3, above TLV). The mean concentration of airborne styrene in G1 group was $10.93{\pm}11.33ppm$, and those of urinary mandelic acid(MA) and phenylglyoxylic acid(PGA) were $0.17{\pm}0.21$ and $0.13{\pm}0.11g/g$ creatinine, respectively. The mean concentration of airborne styrene in G2 and G3 groups were $47.54{\pm}22.43$ and $65.33{\pm}33.47ppm$, respectively, and levels of urinary metabolites such as MA and PGA increased considerably as expected with the increase in exposure level of styrene. The airborne styrene concentration were significantly correlated to the urinary concentration of MA(r=0.784, p=0.000) and PGA(r=0.626, p<0.001). In the 2D electrophoresis, the concentration of five proteins including complement C3 precursor, alpha-1-antitrypsin(AAT), vitamin D binding protein precursor(DBP), alpha-1-B-glycoprotein(A1BG) and inter alpha trypsin inhibitor(ITI) heavy chain-related protein were significantly altered in workers exposed to styrene compared with controls. While expression of complement C3 precursor and AAT increased by exposure to styrene, expression of DBP, A1BG and ITI heavy chain-related protein decreased. These results suggest that the exposure of styrene might affects levels of plasma proteinase, carriers of endogenous substances and immune system. In particular, increasing of AAT with the increase in exposure level of styrene can explain the tissue damage and inflammation by the imbalance of proteinase/antiproteinase and decrease of DBP, A1BG and ITI heavy chain-related protein in workers exposed to styrene is associated with dysfunction and/or declination in immune system and signal transduction