• Journal of Microbiology and Biotechnology
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• v.25 no.11
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• pp.1944-1953
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• 2015

A novel alkaline protease from Streptomyces sp. M30, SapHM, was purified by ammonium sulfate precipitation, hydrophobic interaction chromatography, and DEAE-Sepharose chromatography, with a yield of 15.5% and a specific activity of 29,070 U/mg. Tryptic fragments of the purified SapHM were obtained by electrospray ionization quadrupole time-of-flight mass spectrometry. Nucleotide sequence analysis revealed that the gene sapHM contained 1,179 bp, corresponding to 392 amino acids with conserved Asp156, His187, and Ser339 residues of alkaline protease. The first 24 amino acid residues were predicted to be a signal peptide, and the molecular mass of the mature peptide was 37.1 kDa based on amino acid sequences and mass spectrometry. Pure SapHM was optimally active at 80℃ in 50 mM glycine-NaOH buffer (pH 9.0), and was broadly stable at 0-50℃ and pH 4.0-9.0. The protease relative activity was increased in the presence of Ni²⁺, Mn²⁺, and Cu²⁺ to 112%, 113%, and 147% of control, respectively. Pure SapHM was also activated by dimethylformamide, dimethyl sulfoxide, Tween 80, and urea. The activity of the purified enzyme was completely inhibited by phenylmethylsulfonyl fluoride, indicating that it is a serine-type protease. The K_m and V_max values were estimated to be 35.7 mg/ml, and 5 × 10⁴ U/mg for casein. Substrate specificity analysis showed that SapH was active on casein, bovine serum albumin, and bovine serum fibrin.

• 한국생물공학회:학술대회논문집
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• 2005.04a
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• pp.327-333
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• 2005

Dihydroxy-acid dehydratase (DHAD, 2,3-dihydroxy-acid hydrolyase, EC 4.2.1.9) is one of the key enzymes involved in the biosynthetic pathway of the branched chain amino acid isoleucine and valine. Although the enzyme have been purified and characterized in various mesophiles including bacteria and eukarya, the biochemical properties of DHAD has bee not yet reported from hyperthermophilic archaea. In this study, we cloned, expressed, and purified a DHAD homologue from the thermoacidophilic archaeon Sulfolobus solfataricus P2, which grows optimally at $80\;^{\circ}C$ and pH 3, in E. coli. Characterization of the recombinant S. solfataricus DHAD (rSso_DHAD) revealed that it is the dimeric protein with a subunit molecular weight of 64,000 Da in native structure. rDHAD showed the highest activity toward 2,3-dihydroxyisovaleric acid among 17 aldonic acid substrates Interestingly, this enzyme also displayed 50 % activities toward some pentonic acids and hexonic acids when compared with the activity of this enzyme to the natural substrate. Moreover, rSso_DHAD indicated relatively higher activity toward D-gluconate than any other hexonic acids tested in substrates. $K_m$ and $V_{max}$ values of rSso_DHAD were calculated as $0.54\;{\pm}\;0.04\;mM$ toward 2,3dihydroxyisovalerate and $2.42\;{\pm}\;0.19\;mM$ toward D-gluconate, and as $21.6\;{\pm}\;0.4\;U/mg$ toward 2,3-dihydroxyisovalerate and $13.8\;{\pm}\;0.4\;U/mg$ toward D-gluconate, respectively. In the study for biochemical properties, the enzyme shows maximal activity between $70^{\circ}C$ and $80^{\circ}C$, and the pH range of pH 7.5 to 8.5. The half life time at $80^{\circ}C$ was 30 min. A divalent metal ion, $Mn^{2+}$, was only powerful activators, whereas other metal ions made the enzyme activity reduced. $Hg^{2+}$, organic mercury, and EDTA also strongly inhibited enzyme activities. Particularly, the rSso_DHAD activity was very stable under aerobic condition although the counterparts reported from mesophiles had been deactivated by oxygen.

• Journal of the Korean Society of Food Science and Nutrition
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• v.31 no.5
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• pp.775-781
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• 2002

The T-type amino acid transporter 1 (TATI) is a Na$^{+}$-independent amino acid transporter which selectively trans- ports aromatic amino acids subserving the amino acid transport system T. To understand the transport properties of aromatic amino acids by human TAT1 (hTATl ), we have examined the hTATl -mediated aromatic amino acid transports using a Xenopus laeuis oocyte expression system. When expressed in Xenopin laeuis oocytes, hTATl induced L- [$^{14}$ C]tryptophan transport which was not dependent on Na$^{+}$ or Cl$^{[-10]}$ in the medium. Uptake was time-dependent and exhibited a linear dependence on incubation time up to 30 min. The L- ($^{14}$ C)tryptophan uptake was highly inhibited by L-isomers of tryptophan, tyrosine and phenylalanine, whereas other L-amino acids did not inhibit hTATl -mediated L- ($^{14}$ C)tryptophan uptake. The hTATl induced the relatively low-affinity transport of aromatic amino acids such as L- ($^{14}$ C)tryptophan, L- ($^{14}$ C)tyrosine and L- ($^{14}$ C)phenylalanine (Km values: 450～750 $\mu$M), consistent with the properties of classical amino acid transport system T. The L- ($^{14}$ C)tryptophan uptake did not show any remarkable pH dependence within the pH range of 5.5 to 8.5. The time-dependent efflux of L- ($^{14}$ C)tryptophan was detected from the oocytes expressing hTATl, which was not affected by the presence or absence of L-tryptophan in the extracellular medium, indicating that hTATl-mediated transport is due to the facilitated diffusion. Expression of hTATl in Xenopu laevis oocytes induced the transport of tryptophan, tyrosine and phenylalanine, indicating that hTATl is a transporter subserving system T These results suggest that hTATl has essential roles in the absorption of aromatic amino acids from epithelial cells to the blood stream. Hecause hTATl is proposed to be crucial to the efficient absorption of aromatic amino acids from intestine and kidney, its defect such as blue diaper syndrome could be involved in the disruption of aromatic amino acid transport.ort.

• 한국지구물리탐사학회:학술대회논문집
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• 2005.09a
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• pp.125-153
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• 2005

It has been widely known that the seismic piezo-cone penetration test (SCPTU) is one of the most useful techniques for investigating the geotechnical characteristics including dynamic soil properties. As the practical applications in Korea, SCPTU was carried out at two sites in Busan and four sites in Incheon, which are mainly composed of alluvial or marine soil deposits. From the SCPTU waveform data obtained from the testing sites, the first arrival times of shear waves were and the corresponding time differences with depth were determined using the cross-over method, and the shear wave velocity profiles (VS) were derived based on the refracted ray path method based on Snell's law and similar to the trend of cone tip resistance (qt) profiles. In Incheon area, the testing depths of SCPTU were deeper than those of conventional down-hole seismic tests. Moreover, for the application of the conventional CPTU to earthquake engineering practices, the correlations between VS and CPTU data were deduced based on the SCPTU results. For the empirical evaluation of VS for all soils together with clays and sands which are classified unambiguously in this study by the soil behavior type classification Index (IC), the authors suggested the VS-CPTU data correlations expressed as a function of four parameters, qt, fs, $\sigma$, v0 and Bq, determined by multiple statistical regression modeling. Despite the incompatible strain levels of the down-hole seismic test during SCPTU and the conventional CPTU, it is shown that the VS-CPTU data correlations for all soils clays and sands suggested in this study is applicable to the preliminary estimation of VS for the Korean deposits and is more reliable than the previous correlations proposed by other researchers.

• The Korean Journal of Food And Nutrition
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• v.14 no.1
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• pp.34-39
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• 2001

The purpose of this study was to examine physical properties of the addition of green tea powder on bread flour and dough rheology of white pan bread. Three levels(0.1, 0.5 and 1.0% ) of each green tea powder with bread flour were tested for their effects in dough mixing using rapid disco analyzer, alveogram and farinogram. Addition of green tea powder tended to reduce initial pasting temperature and increase peak viscosity, break down and set back. L(extensibility) and G(swelling index) value in alveogram showed decrement with increasing green tea powder. These meant that the volume of white pan bread would show same tendency. The use of green tea powder increased consistency and water absorption of the bread flour but decreased development time, salability and degree of softening on farinogram. White pan bread with green tea powder had higher value of hardness and springness than without it. Sensory evaluation determined that the white pan bread with 0.5% green tea powder had the highest score.

• ANNALS OF ANIMAL RESOURCE SCIENCES
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• v.30 no.3
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• pp.111-120
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• 2019

The objective of this study was to investigate the effect of different feed inoculation method on rumen fermentation in an in vitro. Three experimental treatments were used: control (CON, direct dispersion of feed (2 g) in rumen fluid), combinations of direct dispersion (1 g) and nylon bag (DNB, pore size: 50 ㎛, 1 g), and nylon bag (NB, 2 g). An in vitro fermentation experiment was carried out using strained rumen fluid for 48 h incubation time and timothy was used as a substrate. At the end of the incubation, in vitro dry matter digestibility (IVDMD), in vitro neutral detergent fiber digestibility (IVNDFD), pH, volatile fatty acids (VFA), ammonia nitrogen (NH₃-N), and microbial community were evaluated and gas production was estimated at 3, 6, 12, 24, 48 h incubation periods. Gas production was higher in CON than DNB and NB at 6 and 12 h incubation time (p<0.01). There were no differences in final gas production, pH, NH₃-N concentration, total VFA production, and VFA profiles among treatments. The IVDMD was lowest in CON (p<0.01) but the IVNDFD was not differed by feed distribution methods. There were no significant differences in general bacteria and fungi. Protozoa count was highest in NB treatment among treatments (p<0.01). The abundance of cellulolytic bacteria, Ruminococcus flavefaciens and Fibrobacter succinogenes, was highest in the CON among treatments (p<0.01).

• Tuberculosis and Respiratory Diseases
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• v.46 no.6
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• pp.795-802
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• 1999

Objective : Cardiopulmonary exercise test is a useful tool to evaluate the operative risk and to plan exercise treatment for the patients with chronic obstructive pulmonary disease(COPD). In cardiopulmonary exercise test, most of the measured parameters are recorded at the time of peak exercise, which are hard to attain in COPD patients. So we evaluated the usefulness of the parameter, breathing reserve index(BRI=minute ventilation [$V_E$]/maximal voluntary ventilation[MVV]) at the time of anaerobic threshold($BRI_{AT}$) for the differentiation of COPD patients with normal controls. Methods : Thirty-six COPD patients and forty-two healthy subjects underwent progressive, incremental exercise test with bicycle ergometer upto possible maximal exercise. All the parameters was measured by breath by breath method. Results : The maximal oxygen uptake in COPD patients (mean$\pm$SE) was $1061.2{\pm}65.6ml/min$ which was significantly lower than $2137.6{\pm}91.4ml/min$ of normal subjects(p<0.01). Percent predicted maximal oxygen uptake was 54.3% in COPD patients and 86.0% in normal subjects(p<0.01). Maximal exercise(respiratory quotient; $VCO_2/VO_2{\geq}1.09$) was accomplished in 7 of 36 COPD patients(19.4%) and in 18 of 42 normal subjects(42.9%). The $BRI_{AT}$ of COPD patients was higher($0.50{\pm}0.03$) than that of control subject($028{\pm}0.02$, p<0.01), reflecting early hyperventilation in COPD patient during exercise. The correlation between $BRI_{AT}$ and BRI at maximal exercise in COPD patients was good(r=0.9687, p<0.01). Conclusion : The $BRI_{AT}$ could be a useful parameter for the differentiation of COPD patients with normal controls in the submaximal cardiopulmonary exercise test.

• Journal of Yeungnam Medical Science
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• v.17 no.1
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• pp.21-30
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• 2000

Background: The purpose of the present study was to investigate the effect of exercise on the activities of antioxidant enzymes, super oxide dismutase(SOD), glutathione peroxidase(GPX) and catalase(CAT) of skeletal muscle(gastrocnemius) and liver in streptozotocin(STZ) induced diabetic rats. The malondialdehyde(MDA) concentration was also measured as an index of lipid poroxidation of tho tissues by exercise-induced oxidative stresses in diabetic rats. Material and Methods: Male Sprague-Dawley rats were randomly divided into control and STZ-induced diabetic rats. The STZ in citrate buffer solution was injected twice at S days intervals intraperitoneally(50, 70 mg/kg respectively). On the 28th day after the first STZ injection, the diabetic animals were randomly divided into pre- and post-exercise groups, The exercise was introduced to the rats of post-exercise group by treadmill running until exhaution with moderate intensity ($V_{O2max}$: 50-70%) of exercise. The duration of average running time was 2 hours and 19 minutes. Results: The blood glucose concentration was increased(p<0.001) and plasma insulin concentration was decreased(p<0.001) in the diabetic rats. The glycogen concentration in the muscle and liver was decreased by exhaustive exercise in the diabetic rats(p<0.001), In the skeletal muscle, the activities of GPX was increased(p<0.05) and the activities of SOD and CAT were not changed in the diabetic rats compare to those of the control rats. The activities of GPX was not changed by exercise but the activities of SOD(p<0.01) and CAT(p<0.01) were decreased by exercise in the diabetic rats, The concentration of MDA was not changed by exercise in diabetic rats, and the values of pre-exercise and post-exercise diabetic rats were not different from the value those of control rats, In the liver, the activities of SOD was decreased(p<0.01), and the activities of GPX and CAT were not changed in diabetic rats compared to the values of control rats, The activities of SOD, GPX and CAT were not changed by exercise in diabetic rats but the activity of SOD seemed to decrease slightly, The MDA concentration was increased in the diabetic rats compared to the values of control rats(p<0.001), but there was no change of MDA concentration by exercise in diabetic rats, Conclusions: In summary, exhaustive physical exercise did not seem to impose oxidative stress on the skeletal muscle because of due to oxygen free radicals, regardless of the decrease in SOD and CAT in the diabetic rats, In liver tissue, the tissue damage by oxidative stress was observed in diabetic rats but the additional tissue damage by exhaustive physical exercise was not observed.