• Applied Biological Chemistry
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• v.48 no.3
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• pp.267-272
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• 2005

trans-Cinnamaldehyde, a major component of Cinnamomum cassia, was quantitatively analyzed using the $^1H-NMR$ spectrometry. Applicability of this method was confirmed through observing the variation of chemical shift in the $^1H-NMR$ spectrum of t-cinnamaldehyde and the integration value according to various sample concentrations or running temperatures. When the $^1H-NMR$ spectrometry was run for t-cinnamaldehyde (7.1429 mg/ml) at 19, 25, 30, 40 and $50^{\circ}C$, the chemical shifts of the doublet methine signal due to an aldehyde group were observed at 9.7202, 9.7184, 9.7169, 9.7142 and 9.7124 ppm, respectively, to imply that the running temperature had no significant variation in the chemical shift of the signal. The integration values of the signal were $1.37\;(19^{\circ}C),\;1.37\;(25^{\circ}C),\;1.37\;(30^{\circ}C),\;1.37(40^{\circ}C)$ and $1.37(50^{\circ}C)$, respectively, to also indicate running temperature gave no effect on the integration value. When the sample solutions with various concentrations such as 0.4464, 0.8929, 1.7857, 3.5714, 7.1429 and 14.286 mg/ml were respectively measured for the $^1H-NMR$ at $25^{\circ}C$, the chemical shifts of the aldehyde group were observed at 9.7206, 9.7201, 9.7196, 9.7192, 9.7185 and 9.7174 ppm. Even though the signal was slightly shifted to the high field in proportion to the increase of sample concentration, the alteration was not significant enough to applicate this method. The calibration curve for integration values of the doublet methine signal due to the aldehyde group vs the sample concentration was linear and showed very high regression rate ($r^2=1.0000$). Meantime, the $^1H-NMR$ spectra (7.1429 mg/ml $CDCl_3,\;25^{\circ}C$) of t-cinnamaldehyde and t-2-methoxycinnamaldehyde, another constituent of Cinnamomum cassia, showed the chemical shifts of the aldehyde group as ${\delta}_H$ 9.7174 (9.7078, 9.7270) for the former compound and ${\delta}_H$ 9.6936 (9.6839, 9.7032) for the latter one. The difference of the chemical shift between two compounds was big enough to be distinguished using the NMR spectrometer with 0.45 Hz of resolution. The contents of cinnamaldehyde in Cinnamomum cassia, which were respectively extracted with n-hexane, $CHCl_3$, and EtOAc, were determiend as 94.2 \;mg/g (0.94%), 137.6 mg/g (1.38%) and 140.1 mg/g(1.40%) t-cinnamaldehyde in each extract, respectively, by using the above method.

• Applied Biological Chemistry
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• v.18 no.4
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• pp.194-202
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• 1975

This study was conducted to study the properties of the water-soluble natural chelating agents from garbage compost and activated sewage sludge responsible for Fe chelation, which is closely associated with the effectiveness in correcting iron chlorosis in plant. The water-soluble fraction of these materials was fractionated by menas of Sephadex gel filtration and the fractions of Fe chehates were traced by radioactive $^{59}Fe$. The fractions were examined by ultraviolet and infrared. spectroscopy and stability constants for Fe. The water-soluble fraction from garbage compost was separated by Sephadex G-25 into approximately four fractions. Most of the added $^{59}Fe$ was associated with fraction I, which appeared at the void volume. Further fractionation by Sephadex G-50 indicated that the molecular weight of water-soluble chelating agents is in the approximate range of 5000 to 10,000. The water-soluble fraction from activated sewage sludge gave six fractions by Sephadex G-25. Most of the added $^{59}Fe$ was found in the fraction I,II, and III, The molecular weights of most chelating agents associated with $^{59}Fe$ appeared to be less than 5,000 and those of fraction I that appeared at the void volume was in the range of 5,000 to 1,000. Discrepancy between radio activity count and UV absorption indicated the heterogeneity of the fractions obtained by Sephadex gel filtration. Ultraviolet absorption spectra of all fractions separated by Sephadex G-25 and containing chelating agents showed no differences. Fraction IV and V of sewage extract showed absorption maxima and shifting similar to nucleic acid components suggesting the presence of decomposition products of nucleic acid. Similarity fraction VI contained phenolic type amino acid groups. Fraction I of compost extract contained most of the added $^{59}Fe$ and showed weak but extra definite absorption in the 1230, and $1270cm^{-1}$ region, suggesting that extra oxygen groups in polyphenolic structure were probably involved in Fe chelation. In sewage extract, fraction I,II, and III in which most of the $^{59}Fe$ was found, showed strong definite polypeptide absorption in the region of $1540cm^{-1}$ due to NH deformation and C-N stretching of amide groups in the peptidebond. These extra functional groups in fraction I, II, and III appeared to be associated with Fe chelation. The other fractions, not associated with $^{59}Fe$, still have carboxyl and hydroxyl groups, suggesting that these functional groups in these water extracts may not independently form the Fe chelates. Precipitation of ferric hydroxide precluded measuring the stability constants for Fe-chelates. However, the formation constants for Zn chelates as log K values for compost extract and sewage extract at pH 4.0 from which the strength of chelation with Fe could be presumed, were 8.23, and 9.75, respectively, indicating strong complexation with metals. The chelating capacity of compost extract containing 6.5 g organic matter per liter was 0.82 mM, and that of sewage extract containing 5.3 g per liter was 0. 64 mM.

• Applied Biological Chemistry
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• v.13 no.2
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• pp.111-129
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• 1970

As a series of studies on the nucleic acids and their related substances 210 samples were collected from 76 places such as farm soil, compost of heap, nuruk and meju to obtain microbial strains which produce 5'-phosphodiesterase. From these samples total of 758 strains were isolated by the use of dilution pour plate method. For all isolated strains primary screening of the productivity of RNA depolymerase was performed and useful strains with regard to 5'-phosphodiesterase productivities were identified. For these useful strains optimum condition, the effect of various compounds on the activity of 5'-phosphodiesterase, and the optimum condition for enzyme reaction were discussed. The quantitative of 5'-mononucleotides produced by the action of 5'-phosphodiesterase was performed using anion-exchange column chromatography and their identified was done by paper chromatography, thinlayer chromatography, ultra violet spectrophotometry, and characteristic color reaction using carbazole and schiff's reagent. (1) Penicillium citreo-viride PO 2-11 and Streptomyces aureus SOA 4-21 from soil were identified as a potent 5'-phosphodiesterase producing strains. (2) Optimum culture conditions for Penicillium citreo-viride PO 2-11 strain isolated were found to be pH 5.0 and $30^{\circ}C$, and the optimum conditions for enzyme action of 5'-phosphodiesterase were pH 4.2 and $60^{\circ}C$. Best carbon source for the production of 5'-phosphodiesterase was found to be sucrose and ammonium nitrate for nitrogen source. Addition of 0.01% corn steep liquor or yeast extract exhibited 20% increase in the amount of 5'-phosphodiesterase production compared to the control. 5'-phosphodiesterase produced by this strain was activated by $Mg^{++},\;Ca^{++},\;Zn^{++},\;Mn^{++}$ and was inhibited by EDTA, citrate, $Cu^{++},\;CO^{++}$. 5'-phosphodiesterase produced 5'-mononucleotide from RNA at a rate of 65.81%, and among the 5'-mononucleotides accumulated 5'-GMP only was found to have flavorous and the strain was also found lack of 5'-AMP deaminase. Productivity of flavorous 5'-GMP was found to be 186.7mg per gram of RNA. (3) Optimum culture canditions for the isolated Streptomyces aureus SOA 4-21 strain were pH 7.0 and $28^{\circ}C$, and the optimum conditions for the action of 5'-phosphodiesterase were pH 7.3 and $50^{\circ}C$. The best carbon source for 5'-phosphodiesterase production was found to be glucose and that of nitrogen was asparagine. Addition of 0.01% yeast extract exhibited increased productivity of 5'-phosphodiesterase by 40% compared to the non-added control. 5'-phosphodiesterase produced by this strain was activated by $Ca^{++},\;Zn^{++},\;Mn^{++}$ and was inhibited by citrate, EDTA, $Cu^{++}$. It was also found that the strain produce 5'-AMP deaminase in addition to 5'-phosphodiesterase. For this reason although decomposition rate was 63.58% the accumulation of 5'-AMP, 5'-CMP, 5'-GMP and 5'-UMP occurred by the breakdown of RNA. In the course of these reaction 5'-AMP deaminase converted 60% of 5'-AMP thus produced into 5'-IMP and flavorous 5'-mono nucleotide production was significantly increased by this strain over the above mentioned one. Production rates were found to be 171.8mg per grain of RNA for 5'-IMP and 148.2mg per gram of RNA for 5'-GMP, respectively.

• The Korean Journal of Ecology
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• v.22 no.3
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• pp.145-153
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• 1999

Authors report the changes of the community structure and secondary succession after fire in Mt. Samma (333 m). Approximately six ha of the red pine (Pinus densiflora) forest and its floor vegetation were burned and cutted down. The vegetation and soil properties were investigated in the burned and unburned sites from April to October, 1998. The dominant species based on SDR₃ in the burned site were Lespedeza cyrtobotrya (96.87), Quercus serrata (77.90), Cyperus amuricus (46.22) and Miscanthus sinensis var. purpurascens (38.33), whereas the dominant species in the unburned site were Pinus densiflora (100.00) and Q. variabilis (66.10) at the tree layer, Q. serrata (100.00) and Zanthoxylum schinifolium (29.64) at the shrub layer., and Q. serrata (76.30) and M. sinensis var. purpurascens (72.84) at the herb layer. The biological spectra based on SDR₃ were Th-D₁-R/sub 5/-e for the burned and H (M)-D₁-R/sub 5/-e type for the unburned site, respectively. The index of similarity (CCs) between the burned and unburned sites was 0.41. The degree of succession (DS) were 609 for the burned and 1168 for the unburned site, respectively. The species diversity (H) and evenness indices (e) of the burned site were lower than those of the unburned site, but the dominance indices (C) was higher in the burned site. In the analysis of soil properties, pH, the content of NO₃/sup -/-N, available phosphrous, and exchangeable cations (K/sup +/, Ca/sup 2+/, Na/sup +/, Mg/sup 2+/) in the burned site were higher than those in the unburned site, whereas the contents of organic matter, total carbon, total nitrogen and NH₄/sup +/-N were lower in the burned site. The results show that dominant species in the burned site change from Miscanthus-Lespedeza→Lespedeza→Quercus and finally to Quercus forest, and the changes of soil properties also affect the early vegetational succession after fire.

• Progress in Medical Physics
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• v.19 no.2
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• pp.95-101
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• 2008

In this study, we observed the alteration of choline signal intensity in hippocampus region of the depressive rat model induced by forced swimming test (FST). The purpose of this study was to evaluate the antidepressant efficacy in the depressive animal model using MR spectroscopy. Fourteen experimentally naive male Sprague-Dawley rats weighting $160{\sim}180\;g$ were used as subjects. Drug injection group was exposed to the FST except for control group. The drugs were administered subcutaneously (SC) in a volume equivalent to 2ml/kg. And three injections were administered 23, 5, and 1h before beginning the given test. 1H MR spectra were obtained with use of a point resolved spectroscopy (PRESS) localization sequence performed according to the following parameters: repetition time, 2500 ms; echo time, 144 ms; 512 average; 2048 complex data points; voxel dimensions, $1.5{\times}2.5{\times}2.5\;mm^3$ ; acquisition time, 25min. There were no differences in NAA/Cr and Cho/Cr ratio between the right and the left hippocampus both normal control rats and antidepressant-injected rats. Also, no differences were observed in NAA/Cr and Cho/Cr ratio between the normal control rats and the antidepressant-injected rats both the right and the left hippocampus. In this study, we found the recovery of choline signals in the depressive animal model similar to normal control groups as injecting desipramine-HCl which was antidepressant causing anti-immobility effects. Thus, we demonstrated that MR spectroscopy was able to aid in evaluating the antidepressant effect of desipramine-HCl.

• Journal of Life Science
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• v.19 no.9
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• pp.1218-1225
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• 2009

Difractose anhydrides (DFAs) is studied as a sweetener for diabetics because of its structural property. DFAs have four types: DFA I, III, IV (degradation of levan) and V (degradation of inulin). Especially, DFA IV has been shown to enhance the absorption of calcium in experiments using rats. Levan fructotransferase is an enzyme for producing di-d-fructose-2,6':6,2-dianhydride (DFA IV). To identify structural characterization, we purified wild-type and mutants (D63A, D195N and N85S) of levan fructotransferase (LFTase) from Microbacterium sp. AL-210. These proteins were purified to apparent homogeneity by Ni-NTA affinity column, Q-sepharose ion exchange and gel filtration chromatography and detected by SDS-PAGE. They were also analyzed by circular dichroism (CD) measurements, JNET secondary structure prediction, activity measurements at various temperatures, and pH analysis. The optimum pH for the enzyme-catalyzed reaction was pH 7.5 and optimum temperature was observed at $55^{\circ}C$. Along with wild-type LFTase, mutants were analyzed by CD measurement, fluorescence analysis and differential scanning calorimetry (DSC). N85S showed less $\alpha$-helix and more $\beta$ strand than others. Also, N85S showed almost the same curve as wild-type in their steady-state fluorescence spectra, whereas mutant D63A and D195N showed higher intensity than wild-type. The amino acid sequence of wild-type LFTase was compared to the sequences of exo-inulinase from Aspergillus awamori, a plant fructan 1-exohydrolase from Cichorium intybus, and Thermotogo maritime (Tm) invertase and showed a high identity with Exo-inulinase from Aspergillus awamori.