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Bacillus licheniformis KFB-C14가 생산하는 내열성 Chitinase의 정제 및 특성

  • Hong, Bum-Shik;Yoon, Ho-Geun;Shin, Dong-Hoon;Cho, Hong-Yon
    • Microbiology and Biotechnology Letters
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    • v.24 no.5
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    • pp.567-573
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    • 1996
  • Chitinase (EC 3.2.1.14) from culture fluid of Bacillus licheniformis KFB-C14 was purified 66-folds to homogenity in overall yield of 21% by ammonium sulfate fractionation, DEAE-Toyopearl, Butyl-Toyopearl and TSK-Gel HW-55F column chromatography. The enzyme protein had a molecular weight of about 86,000 and was composed of one subunit. The enzyme was significantly stable not only at high temperature but also on treatment with organic solvents and protein denaturants such as SDS, urea and guanidine-HC1. The optimum temperature and pH for reaction was 60$\circ $C and 6.0, respectively. The enzyme activity was inhibited by only Mn$^{2+}$ ion, but not inhibited by EDTA, N- ethylmaleimide and pCMB. The enzyme had high activity with colloidal chitin (V$_{max}$: 421) and commercial chitin (V$_{max}$: 480), but not with typical substrates of exo type chitinase. The thermostable chitinase had an useful reactivity for producing functional chitooligosaccharide, showing the production of (GlcNAc)$_{1}, (GlcNAc)$_{3}$, and (GlcNAc)$_{2}$ as major product.

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Characteristic of Alcohol Fermentation by the Culture of High Cell Density (Functional Relationship among Specific Growth Rate, Sugar Concentration, Cell Concentration and Alcohol Concentration) (고농도 균체배양에 의한 알콜발효특성 규명 (비성장속도와 당농도, 균체농도, 알콜농도의 함수관계))

  • 허병기;김형철양지원목영일
    • KSBB Journal
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    • v.6 no.1
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    • pp.85-90
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    • 1991
  • Experiments of alcohol fermentation of the yeast,K. fragi1is CBS 1555 were performed to obtain the following results. In these experiments, the initial concentrations of sugar which was composed of inulin and fructose as weight ratio of one to one were 30, 50, 75, 100 and 150g/l and the initial densities of the microorganism were less than 0.5g/l, 10g/1 to 15g/1, and 50g/l. The functional relationship among specific growthrates, sugar concentrations, and alcohol concentrations could be expressed by Aiba-shoda equation and the specific growth rate represented the trend that decreased with increase in the initial concentration of the microorganism. Also, $\mu$max and Ks of Monod's equation could be expressed as the function of initial cell concentration like the following equations. $\mu$max=0.8-0.008X Ks=0.54X+8 In the region that sugar, alcohol and cell concentrations were 10g/1 to 120g/l, 0g/l to 60g/l and 0.5g/l to 50g/l respectively, the differences between the experimental values and the calculated ones for specific growth rate approached to 40% with respect to experimental values at the worst cases, but in most cases, those were distributed in the range of less than 20%.

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Purification and Properties of $\alpha$-Glucosidase from Mococcus halophilus (Pediococcus halophilus로부터 생성한 $\alpha$-Glucosidase의 정제 및 특성)

  • 민해기;이호근;문지웅;강국희
    • Microbiology and Biotechnology Letters
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    • v.20 no.2
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    • pp.143-149
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    • 1992
  • A bacterial strain No. 2, which highly produced a-glucosidase, was isolated from Kimchi and identified to be a similar species of Pediococcus halophilus. This enzyme was purified by protamine sulfate, ammonium sulfate fractionation, ion exchange and gel filtration. The maximal a-glucosidase activity was observed at pH 6.0 and this enzyme was stable at pH 6.0~ 7.5. The optimum temperature of this enzyme activity was $37^{\circ}C$, but enzyme activity was gradually lost above $37^{\circ}C$. This enzyme was activated by 10 mM MgCh and inhibited by 10 mM mercaptoethanol. The kinetics of PNPG(p-Nitrophenyl-a-D-glucopyranoside) and maltose were Kp0.52 mM/27.5 pg protein, $V_{max}$= 0.021 mM/min 27.5 ${\mu}g$ protein and $K_m$= 0.32 mMD7.5 ${\mu}g$ protein, $V_{max}$= 0.025 mM/min 27.5 ${\mu}g$ protein, respectively. The molecular weight of $\alpha$-glucosidase was about 37, 000.

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Development of a Core Set of Korean Soybean Landraces [Glycine max(L.) Merr.]

  • Cho, Gyu-Taek;Yoon, Mun-Sup;Lee, Jeong-Ran;Baek, Hyung-Jin;Kang, Jung-Hoon;Kim, Tae-San;Paek, Nam-Chon
    • Journal of Crop Science and Biotechnology
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    • v.11 no.3
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    • pp.157-162
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    • 2008
  • A total of 2,765 accessions were used as the initial set having both seed coat color and 100-seed weight data. As a result of molecular profiling using six SSR markers followed by stratification based on their usages, 335 accessions(12.1%) were selected by clustering based on UPGMA. Since 75 out of 335 accessions were mixed in phenotypic traits as a result of characterization, 260 accessions were finally set as a core set. This core set revealed nearly the same diversity compared with the other results on morphological traits of Korean soybean landraces. In total, 115 alleles(19.2 alleles per locus) were detected in the initial set and 79 alleles(13.2 alleles per locus) were detected in the core set. All 30 major alleles were present in the initial set and in the core set as well. In allele coverage, the core set was 71.4% of the initial set. These comparisons of number of alleles, gene diversity and coverage indicated that the core set represented the entire set well.

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Purification and Characterization of the Red Carotenoprotein from the Skin of Ascidian, Halocynthia roretzi 1. Purification and Characterization of the Caritenopritein (멍게껍질로부터 분리제정한 적색 Carotenoprotein의 특성 1. Carotenoprotein 의 정제 및 특성)

  • Kang, Ok-Ju;Suh, Myung-Ja;Lee, An-Jong;Kim, Se-Kwon
    • Journal of Life Science
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    • v.5 no.4
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    • pp.170-180
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    • 1995
  • A carotennnoprotein from the skin of Ascidian(Halocynthia roretzi) was extracted by Triton X-100 and purified by ammonium sulfate fraction, SephadexG-200 charomatography and DEAE-cellulose ion exchange chromatography. The carotenoprotein was redwith broad $\lambda$$_{max}$ between 495, 467 and 318nm. The red carotenoprotein had an approximate molecular weight of 326KDa(gel filtration). SDS-PAGE indicated the presence of two polypeptodes of 84.1KDa and 74.4KDa, with different mobility in polyacrylamide gel electrophoresis. In the presence of denaturing agents such as organic solvent aand extreme pH, the red complex readily disociates to liberate the yellow carotenoid($\lambda$$_{max}$ 452nm) and a colourless apoprotein. The amino acid composition of carotenoprotein were mainly threonine(15.2%), aspartic acid(12.2%), glutamic acid(11.9%) and serine(9.6%), while proline was not found. The carotenoprotein consisted of lipids as structure units. Its major fatty acids composion were C$_{18:1}$, C$_{16:1}$, and C$_{16:0}$. The monounsaturated fatty acids(41.5%) contained abundant content compared to other fatty aacids(polyunsaturated fatty acids 37.4%, saturated fatty acids 20.6%).

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Aerobic Capacity and Ventilatory Response During Incremental Exercise in Elite High School Cyclist (점진부하 운동에서 중고교 엘리트 사이클 선수들의 유산소능력과 폐환기 반응)

  • Lee, Dae-Taek;Bae, Yoon-Jung
    • Journal of Life Science
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    • v.20 no.3
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    • pp.437-443
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    • 2010
  • This study was designed to examine the aerobic capacity and ventilatory response during an incremental exercise in elite high school cyclists. Twelve boys ($17{\pm}1\;yr$, $175{\pm}5\;cm$, $70{\pm}9\;kg$) participated in anthropometric measurements, incremental exercise testing, and pulmonary function tests. During incremental exercise testing using a cycle ergometer, their maximal oxygen uptake ($VO_2max$), maximal power output, ventilation, ventilatory equivalents for oxygen ($V_E/VO_2$) and carbon dioxide ($V_E/VCO_2$), respiratory rate, and tidal volume were measured. Time variables such as inspiratory time (Ti), expiratory time (Te), breathing time (Tb), and inspiratory duty cycle (Ti/Tb), as well as inspiratory flow rate ($V_T$/Ti) were assessed. Pulmonary function of vital capacity (FVC), forced expiratory volume in one second ($FEV_1$), $FEV_1$/FVC, and peak expiratory flow were evaluated. Their $VO_2max$, maximal heart rate, and Wmax were $57.5{\pm}3.9\;ml{\cdot}kg^{-1}{\cdot}min^{-1}$, $194.1{\pm}8.6\;beat{\cdot}min^{-1}$, and 452 W, respectively. $VO_2max$ was not related to any anthropometric parameters. Most ventilatory variables progressively increased with exercise intensity. As intensity increased, Ti, Tb, Tb decreased while Ti/Tb was maintained. Below an intensity of 250 W, height, weight, body mass index, and body surface were highly correlated with $V_T$/Ti and Ti/Tb (p<0.05). Collectively, $VO_2max$ appeared to be lower than adult cyclists, suggesting a different pattern of ventilatory control as age advances. Morphological characteristics were not related to $VO_2max$ in the population. Time variables of ventilatory response seemed to be related only at an exercise intensity level of less than 250 W. $V_T$/Ti may be related to exercise endurance capacity, but Ti/Tb was similar to adult cyclists.

Purification and Characterization of GTP Cyclohydrolase I from Streptomyces tubercidicus, a Producer of Tubercidin

  • Yoo, Jin-Cheol;Han, Ji-Man;Ko, Ok-Hyun;Bang, Hee-Jae
    • Archives of Pharmacal Research
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    • v.21 no.6
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    • pp.692-697
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    • 1998
  • GTP cyclohydrolase I catalyzing the first reaction in the biosynthesis of pterin moiety of folic acid in bacteria, was purified from Streptomyces tubercidicus by at least 203-fold with a yield of 32% to apparent homogeneity, using ammonium sulfate fractionation, DEAE-cellulose, Sepharose CL-6B, and hydroxylapatite column chromatography. The molecular weight of the native enzyme was estimated to be 230,000 daltons by gel permeation chromatography. The purified enzyme gave a single band on sodium dodesyl sulfate-polyacrylamide gel electrophoresis and its molecular weight was apparently 58,000 daltons. These results indicate that the enzyme consists of four subunits with the same molecular weight. The $K_m$ and $V_{max}$ values for GTP of the purified enzyme were determined to be 80${\mu}$M and 90nmol/min (mg protein), respectively. The optimum pH and temperature for the enzyme reaction were pH 7.5-8.5 and $40-42^{\circ}C$, respectively. Coenzyme or metal ion was not required for the enzyme activity. The enzyme activity was inhibited by most divalent cations, while it was slightly activated by potassium ion. In case of nucleotides, CTP, GMP, GDP, and UTP inhibited enzyme activity, among which GDP exhibited the strongest inhibitory effect.

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The Effect of Isoflavone and/or Grape Seed Oil Supplementation on Blood Lipid Profiles and Bone Strength in Ovariectomized Female Rats (이소플라본과 포도종실유의 보충이 난소절제 흰쥐의 혈중지질 패턴 및 골격강도에 미치는 영향)

  • 정혜경;최창숙;이지현;박원종;강명화
    • Journal of Nutrition and Health
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    • v.36 no.7
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    • pp.667-674
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    • 2003
  • The study was performed to investigate the effects of isoflavone and/or grape seed oil supplementation on serum and liver lipid profiles and bone strength in ovariectomized female rats. Female Spraque-Dawley rats were assigned into 4 groups of ovariectomized (O), isoflavone (0.085 mg/100 g b.w/day) in ovariectomized rats, grape seed oil in ovariectomized rats and isoflavone and/or grape seed oil in ovariectomized rats. After 8 weeks, biochemical profiles of serum, liver and bone were analyzed. Total food intakes, body weight gains and FER (food efficiency ratio) were not statistically significantly different among groups. Total cholesterol, triglyceride and LDL-cholesterol levels in serum were decreased by fed of isoflavone and/or grape seed oils. However, crude lipid and total cholesterol contents in liver were not affect of isoflavone and/or grape seed oil. The hepatic glutathione contents were increased by isoflavone and/or grape seed oil fed. The hepatic glutathione-S-transferase activity in isoflavone and/or grape seed oil supplemented groups were higher than that O group. Bone (scapular and femur bone) dry weight, femur of max weight and bending strength were no significant difference among groups. Our finding suggest that isoflavone/grape seed oils might have potential role for serum lipid profiles improvement and bone strength in vivo.

Effect of Cyclohexane Treatment on the Liver Cytochrome P-450 Dependent Aniline Hydroxylase Activity in Alcohol-pretreated Rats (알코올 전처치한 흰쥐에 Cyclohexane 투여가 Cytochrome P-450 dependent aniline hydroxylase 활성에 미치는 영향)

  • 김병렬;윤종국
    • Journal of Environmental Health Sciences
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    • v.29 no.2
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    • pp.23-28
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    • 2003
  • To evaluate the effect of cyclohexane(CH) treatment on the liver cytochrome P-450 dependent aniline hydroxylase(CYPdAH) activity in alcohol-pretreated animals, CH(1.56 g/kg body weight) was intraperitoneally administered to Sprague-Dawley male rats, which had been drunk 15% alcohol in distilled water for 1,3 and 6 weeks. CH was injected to rats 4 times every other day and the animals were sacrificed at 24 hours after injection of CH. In the alcohol-pretreated rats, liver injuries were not demonstrated on the basis of the liver weight per body weight, the levels of serum alanine aminotransferase and hepatic microsomal glucose-6-phosphatase activities. By the CH treatment, alcohol-pretreated animals showed the significantly increased activity of hepatic microsomal CYPdAH. Concomitantly $V_{max}$ value in CYPdAH was more increased, whereas $K_{M}$ value more decreased in alcohol-pretreated animals by the treatment of CH. In conclusion, the increasing cause of microsomal CYPdAH in CH-treated rats pretreated with alcohol may be due to induction of enzyme protein in rat liver.r.r.

Clinical Application of Ma Huang in the Obesity Treatment (비만처방에서 마황의 임상활용에 대한 고찰;용량, 효과, 부작용 등의 측면에서)

  • Song, Yun-Kyung;Lim, Hyung-Ho
    • Journal of Korean Medicine for Obesity Research
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    • v.7 no.1
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    • pp.1-7
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    • 2007
  • Background : Use of Ma Huang is controversial. Multiple reports of adverse effects led the U.S. Federal Drug Administration (FDA) to announce about prohibiting dietary supplements containing ephedrine alkaloids/Ma Huang. Objective : The objective of the study was to review the evidence from literature and rigorous clinical trials on the dosage, effectiveness, and adverse effects. Methods : Literature searches were conducted in U.S. FDA database after 2003 and digital Uibujeonok(醫部全錄). Hand searches of medical journal and bibliographies of identified articles were conducted. Results : Ma Huang has been used in Korean Oriental Medicine to treatment asthma and other ailments. Recently, Ephedrine/Ma Huang are used for weight loss or to enhance athletic performance in Europe and America, Korea. Ephedrine/Ma Huang are associated with increased risk of psychiatric, autonomic, or gastrointestinal symptoms, and heart palpitations. In addition to that, a risk of serious adverse events, including heart attack, stroke, epilepsy and psychiatric problem are also have been reported. Therefore, max dose of total alkaloids of ephedrine is studied in need of a specialist's part. Conclusion : In most reports, Ma Huang effectively promoted short-term weight loss and enhancement of athletic performance, But, safety with long-term use requires further investigation.

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