• Archives of Pharmacal Research
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• v.23 no.4
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• pp.407-412
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• 2000

The presence of the nitric oxide synthase (NOS) enzyme from Salmonella typhimurium (S. typhimurium) was identified by measuring radiolabeled L-$[^3H]$citrulline and NO, and Western blot analysis. NOS was partially purified by both Mono Q ion exchange and Superose 12HR size exclusion column chromatography, sequentially. The molecular weight of NOS was estimated to be 93.3 kDa by Western blot analysis. The enzyme showed a significant dependency on the typical NOS cofactors; an apparent Km for L-arginine of 34.7 mM and maximum activity between $37^{\circ}C$ and $43^{\circ}C$. The activity was inhibited by NOS inhibitors such as aminoguanidine and $N^{G}$ $N^{G}$-dimethyl-L-arginine. taken together, partially purified NOS in S. typhimurium is assumed to be a different isoform of mammalian NOSs.OSs.

• Interaction and multiscale mechanics
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• v.1 no.3
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• pp.357-368
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• 2008

The result of damage detection from on-site measurements is commonly polluted by unavoidable measurement noises. It is widely recognized that this side influence could be reduced to some extent if the sensor placement was properly designed. Although many methods have been proposed to find the optimal number and location of mono-type sensors, the optimal layout of multi-type sensors need further investigation, because a network of heterogeneous sensors is commonly used in engineering. In this paper, a new criterion of the optimal placement for different types of sensors is proposed. A corresponding heuristic is developed to search for good results. In addition, Monte Carlo simulation is suggested to design a robust damage detection system which contains certain redundancies. The validity of these methods is illustrated by two bridge examples.

• Archives of Pharmacal Research
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• v.20 no.3
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• pp.218-224
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• 1997

Platelet-activating factor (PAF) acetylhydrolase, which removes the acetyl moiety at the sn-2 position, has been found in human amniotic fluid. We purified this enzyme by ammonium sulfate precipitation, and sequential use of DEAE-Sepharose CL-6B, hydroxyapatite, chelating-Sepharose, and Mono Q column chromatographies. This enzyme exhibited broad pH optima and was unaffected by EDTA. Partially purified enzyme had a molecular weight of approximately 34 kDa on SDS-PAGE. In addition, the enzyme activity was inhibited by either diisopropylfluorophosphate(DFP) or p-bromophenacylbromide (p-BPB), suggesting that this enzyme possesses active serine and histidine residues. The enzyme showed similar activity towards PAF and oxidatively modified phosphatidylcholine, but didn't hydrolyze phosphatidylcholine or phosphatidylethanolamine with a long chain fatty acyl group at sn-2 position.

• Journal of Microbiology and Biotechnology
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• v.15 no.5
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• pp.1047-1053
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• 2005

Cyclosporin A binding protein type-19 kDa peptidyl-prolyl cis/trans isomerase (PPIases, EC 5.2.1.8) of Euglena gracilis was purified and some of its biochemical characters were elucidated. Purification of the PPIase was achieved by employing a series of steps involving ammonium sulfate precipitation, Superdex G-75 gel filtration chromatography, Mono­Q anion and Mono-S cation exchange chromatographies, and Superdex S-200 gel filtration chromatography on FPLC. Purified PPIase had a specific activity of 8,250 units/mg, showing a 27-fold increase compared with that of cell-free extract of Euglena gracilis. The enzyme consisted of a single polypeptide chain with a molecular mass of 19 kDa. It showed high substrate specificity to succinyl-Ala-Ala-Pro-Phe-p-nitroanilide, and $k_{car}/K_{m}$, for this substrate was found to be $61.19{\times}10^5/sec$. The isomer distributions were investigated at an equilibrium of seven different peptide substrates, varying Xaa in Suc-Ala-Xaa-Pro-Phe-p-nitroanilide in dimethylsulfoxide. The cis/trans equilibrium constants were estimated to be from 0.14 (Ile) to 0.63 (Gly), which correspond to $12.00\%\;to\;38.52\%$ of the cis population, respectively, under experimental condition. The enzyme was highly sensitive to the immunosuppressive ligand cyclosporin A, but not to other immunosuppressants such as FK506 and rapamycin. Thus, it appears to belong to the class of cyclophilin.

• Korean Journal of Food Science and Technology
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• v.25 no.5
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• pp.576-581
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• 1993

Polygalacturonase(PG) was extracted from Chinese cabbage and partially purified by ammonium sulfate fractionation and ion-exchange chromatography. Three fractions, D-PG, C-1 PG, and C-2 PG, were separated by CM-Sephadex C-25 column chromatography and FPLC Mono Q HR 5/5 or Mono S HR 5/5 column and their physicochemical characteristics were investigated. All three fractions had optimum pH and temperature of 4.5 and $65^{\circ}C$, and were stable in the range of pH $4.5{\sim}8.0$. These fractions were inhibited by 0.8mM $CaCl_2$ or 0.6M NaCl. In the thermal inactivation of PC isozymes at $65{\sim}80^{\circ}C$, z-values were $8.4{\sim}9.3^{\circ}C$ and D-values at $80^{\circ}C$ were In the range of $120{\sim}126s$. Thermodynamic constants were calculated from thermal inactivation data of the isozymes and were applicable to estimation of thermal process time of retort pouched Kimchi.

• Korean Journal of Environment and Ecology
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• v.10 no.1
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• pp.151-159
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• 1996

To investigate the structure and the conservation strategy of natural forest at Sangwonsa, Pirobong, Horyongbong area in Odaesan, 23 plots(20*20m) were set up with random sampling method. Three groups were classified by cluster analysis; Quercus mongolica-Abies holophylla community, Acer mandshuricum- Abies nephrolepis community, and A. holophylla-Ulmus laciniata community. High positive correlations were proved betwween Tilia amurensis and Acer mono; Sorbus commixta and Euonymus macropterus, Betula ermanii, and Viburnum sargentii; E. macropterus and V. sargentii; B. ermanii and V. sargentii; A. mephrolepis and Syringa velutina var. kamibayashii; S. velutina var. kamibayashii and Salix hulteni; Pinus densiflora and A. holophylla. High negative correlations were proved between Q. mongolica and Juglans mandhuricum; Sasa vorealis and E. macropterus; Acer mono and Philadelphus schenckii; A. holophylla and E. macropterus, B. ermanii, S. velutina var. kamibayashii, and Salix hulteni. Species diversity(H') of investigated area was 1.2973~1.46331.

• Journal of Microbiology and Biotechnology
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• v.34 no.3
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• pp.725-734
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• 2024

CYP102A1 from Bacillus megaterium is an important enzyme in biotechnology, because engineered CYP102A1 enzymes can react with diverse substrates and produce human cytochrome P450-like metabolites. Therefore, CYP102A1 can be applied to drug metabolite production. Terpinen-4-ol is a cyclic monoterpene and the primary component of essential tea tree oil. Terpinen-4-ol was known for therapeutic effects, including antibacterial, antifungal, antiviral, and anti-inflammatory. Because terpenes are natural compounds, examining novel terpenes and investigating the therapeutic effects of terpenes represent responses to social demands for eco-friendly compounds. In this study, we investigated the catalytic activity of engineered CYP102A1 on terpinen-4-ol. Among CYP102A1 mutants tested here, the R47L/F81I/F87V/E143G/L188Q/N213S/E267V mutant showed the highest activity to terpinen-4-ol. Two major metabolites of terpinen-4-ol were generated by engineered CYP102A1. Characterization of major metabolites was confirmed by liquid chromatography-mass spectrometry (LC-MS), gas chromatography-MS, and nuclear magnetic resonance spectroscopy (NMR). Based on the LC-MS results, the difference in mass-to-charge ratio of an ion (m/z) between terpinen-4-ol and its major metabolites was 16. One major metabolite was defined as 1,4-dihydroxyp-menth-2-ene by NMR. Given these results, we speculate that another major metabolite is also a mono-hydroxylated product. Taken together, we suggest that CYP102A1 can be applied to make novel terpene derivatives.

• Korean Journal of Plant Resources
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• v.13 no.3
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• pp.227-242
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• 2000

Site characteristics, vegetation structure and dynamics of Kalopanax septemlobus forests in Mt. Heungjeong, Mt. Balwang and Mt. Gariwang were studied to provide the information need for national resources conservation and restoration of K. septemlobus forest in Gangwon-do of Korea. The K. septemlobus forests were distributed at 780m to 1,300m in altitude, steep slope, ridges-slope areas with facing northeast to northwest in good nutrition area. The annual diameter growth of those trees ranged 1.60mm to 2.41mm. Forty-three plots(20$\times$20m) were set up at three natural population of Kalopanax septemlobus forest and TWINSPAN method was used for vegetation structure analysis. The result of TWINSPAN ordination show that geographical features played a role in determining community types in the study areas. Community type of the study areas classified into three groups by TWINSPAN; K. septemlobus-Quercux mongolica- Acer mono community(Mt. Balwang), K. septemlobus-Q. mongolica- Tilia amurensis community(Mt. Heungjeong), K. septemlobus-Betula costata-Q. mongolica community(Mt. Gariwang). In most of the K. septemlobus forests, major woody species appeared predominantly were Q. mongolica, T. amurensis, Acer pseudo-sieboldianum, Lindera obtusiloba, A. mono, Magnolia sieboldii, Ulmus davidiana var. japonica. And also, Those species were surveyed in all the areas. Species diversity(H') of investigated areas ranged 0.3124~0.13992 and this value relatively higher than that of other forests with similar site. The range of similarity indices between surveyed areas was 64.1~64.54%. The distribution pattern of Morista's index show that Kalopanax septemlobus distributed randomly in tree and concentrated a few sample plots in subtree layer and shurb layer. K. septemlobus occupied 37.1~58.8% of tree layer story, 8.4~17.6% of subtree layer, 0~0.38% of shurb layer each surveyed area. The association analysis showed that Kalopanax septemlobus positively associated with Betula ermani, Deutzia glabrata, Carpinus cordata, Fraxinus mandshurica and negatively associated with Symplocos chinensis var. pilosa, Styrax obassia, Acer mono, Euonymus macroptera, Ulmus davidiana var. japonica. According to the diameter distribution of Kalopanax septemlobus community, the community of Mt. Gariwang will gradually decline and replaced by Quercus community. The communities of Mt. Heungjeong and Mt. Balwang has its seedlings and saplings, so it may be sustained.

• Journal of Life Science
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• v.14 no.3
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• pp.435-440
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• 2004

The gene encoding a extremely thermostable 4-$\alpha$-glucanotransferase from a hyperthermophilic archaeon, Thermococcus litoralis, was cloned, sequenced and expressed in Escherichia coli. The amino acid sequence of the enzyme was distantly related to other functionally-related ones, such as D-enzymes. The enzyme is of industrial interest because of a novel activity of producing cycloamylose and is also important for fundamental studies of protein, sugar-metabolizing enzymes. In this paper, the overexpression of 4-$\alpha$-glucanotransferase in E. coli was carried out expression vector system with lac and T7 promoters. The enzyme was successfully overexpressed, and purified by the heat treatment of a cell-free extract, successive Butyl-Toyopearl and Mono Q chromatographies. The purified recombinant enzyme showed the same specific activity and the same mobility in SDS-PAGE as natural enzyme.

• Microbiology and Biotechnology Letters
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• v.30 no.3
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• pp.235-240
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• 2002

Streptomycetes is a group of Gram-positive soil bacteria that growas a branching vegetative mycelium leading to the formation of spores, and display a physiological differenti-ation related to the synthesis of many secondary metabolites including antibiotics. Their complex life cycle and multicellular differentiation require various levels of regulation and types of signal transduction systems including eukaryotic-type serine/threonine protein kinases and prokaryotic-type histidine/aspartic acid protein kinases. Akt kinase that was found in cells is a sorine/threonine kinase controlling signal pathway for multi-tude of important cellular events. The activation or inactivation of Akt kinase in the cell is one of the critical regulatory points to deliver cell proliferation, differentiation, survival or apoptosis signal. To find the regula-tory protein homologous to Akt in Streptomyces, the fluorescien-labeled synthetic peptide (FITC-TRRSR-TESIT) was designed from the consensus sequence of target proteins for Akt kinase. From the difference of the mobility between the nonphosphorylated and phosphorylated synthetic peptides on Agarose gel electro-phoresis, the Akt-phosphorylating activity was monitored. The cell-free extract prepared from Streptomyces griseus IFO 13350 and the Akt homologous protein was purified by ammonium sulfate fractionation and many steps of column chromatographies such as, DEAE-Sepharose, Mono Q, Resource Phenyl-Soporose and Gel permeation column chromatographies. As a result, the protein phosphorylating the fluorescien-labeled Akt substrate was identified and it's molecular weight was estimated as 39 kDa on SDS-PAGE.