• Applied Biological Chemistry
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• v.45 no.4
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• pp.190-194
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• 2002

Expression of Bacillus subtilis glutamyl-tRNA synthetase (GluRS) in Escherichia coli is lethal for the host, probably because this enzyme misaminoacylates ${tRNA_l}^{Gln}$ with glutamate in vivo. In order to overexpress B. subtilis GluRS, encoded by the gltX gene, in E. coli, this gene was amplified from B. subtilis 168 chromosomal DNA using PCR method and the entire coding region was cloned into a pET11a expression vector so that it was expressed under the control or the T7 Promoter. The resulting recombinant pEBER plasmid was transformed into E. coli Novablue (DE3) bearing the T7 RNA polymerase gene for expression. After IPTG treatment, the overproduced enzyme was purified using ammonium sulfate fractionation, Source Q anion exchange chromatography, Superdex-200 gel filtration, and Mono Q anion exchange chromatography. The purified enzyme yielded 18-fold increase in specific activity over the crude cell extract and its molecular weight was approximately 55 kDa on SDS-PAGE.

• Biomedical Science Letters
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• v.6 no.4
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• pp.261-268
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• 2000

Fibrinolytic enzyme (FE-2) was purified from the fruiting bodies of Tricholoma saponaceum using DEAE-Cellulose chromatography and Mono-S column chromatography, The enzyme has a molecular weight of 18.23 kDa and include Zn$^{2+}$ ion as found by ICP/MS. The N-terminal amino acid sequence of the enzyme was A-L-Y-V-G-X-S-P-X-Q-Q-S-L-L-V It has a pH optimum at pH 7.5, suggested that FE-2 was a neutral pretense. The activity of FE-2 was highly inhibited by EDTA and 1,10-phenanthroline, indicating that the enzyme is a metalloprotease. The activity of FE-2 was increased by $Mg^{2+}$, Zn$^{2+}$, Fe$^{2+}$, and Co$^{2+}$, but the enzyme activity was totally inhibited by Hg$^{2+}$. No inhibition was found with PMSF, E-64, pepstatin and 2-mercaptoethanol. The enzyme hydrolyzed both $A\alpha$ and B$\beta$ chains of human fibrinogen. The $\gamma$ chain was resistant to hydrolysis by FE-2.

• Journal of Life Science
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• v.8 no.2
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• pp.181-188
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• 1998

Two distinct ADP-ribosyltransferases, termed Yac-1 and Yac-2 from mouse lymphoma cells were recently cloned and characterized. Yac-1 enzyme possesses ADP-ribosyltransferases activity. In contrast, Yac-2 has significant NAD glycohydrolase activity and may preferentially hydrolyze NAD. Yac-2 possesses a glutamate at position 219 adjacent to the two consdrved glutamic acid residues. To study the effect of Glu-219 on enzyme activities, Glu-219 was mutagenized to Glutamine (E219Q) or alanine (E219A) using a two-step recombinant polymerase chain reaction procedure. Replacing Glu at position 219 with Gln or Ala resulted in 56 (E219Q) or 66% (E219A) reduction in ADP-ribosyltranferase activity. The NAD glycohydrolase activity of Yac-2 protein were not altered by the mutations. These results indicate that Glu-219 in Yac-2 enzyme plays an important role in ADP-ribosyltransferase, but not NAD glycohydrolase activity.

• Journal of Ecology and Environment
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• v.29 no.3
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• pp.265-275
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• 2006

This study represents the mechanism of progressive succession and potential natural vegetation on the forest vegetation in and surrounding Daegu. As a result of DCA, the feature of community was determined by an altitude and humid gradients. The soil moisture, contents of organic matter and total nitrogen increased as the community developed. In the interspecific association analysis, the forest vegetation was divided into two species groups and they were influenced by temperature and soil moisture. Especially, each two groups showed different stages of vegetation development according to the progressive succession and life form composition supported those results. It was predicted that Quercus variabilis, Q. acutissima, Q. dentata and Pinus densiflora communities would develop into Q. serrata community or Q. mongolica community depending on their location or species composition. In the study area, the potential natural vegetation was divided into 3 communities by biogeographical gradients such as species composition, soil environment, and geographical features: 1)Q. mongolica community in the middle-upper area of the mountain, 2)Q. serrata community in the middle-lower area of the mountain and 3)Carpinus cordata-Acer mono community in the cove area. It is suggested that the Q.mongolica and C.cordata-A.mono communities become actual vegetation and potential natural vegetation. But it is also suggested that the P. densiflora community would be changed into the potential natural vegetation of the Q. mongolica community and Q. serrata community on the basis of the present species composition.

• Journal of Microbiology
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• v.38 no.3
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• pp.187-191
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• 2000

A validation study was conducted to determine the efficacy of solvent/Detergent (S/D) inactivation and Q-Sepharose column chromatographic removal of the human immunodeficiency virus (HIV) during the manufacturing of a high purity antihemopilic factor VIII (GreenMono) from human plasma. S/D treatment using the organic solvent, tri (n-butyl) phosphate, and the detergent, Trition X-100, was a robust and effective step in eliminating HIV-1. The HIV-1 titer was reduced from an initial titer of 8.3 log10 TCID50 to undetectable levels within one minute of S/D treatment, HIV-1 was effectively partitioned form factor VIII during Q-Sepharose column chromatography with the log reduction factor of 4.1 . These results strongly assure the safety of GreenMono From HIV.

• Microbiology and Biotechnology Letters
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• v.29 no.4
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• pp.248-252
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• 2001

Recombinant Human serum albumin (rHSA) was purified to near homogeneity from H, polymorpha using heat treatment, ultrafiltratipn and Phenyl Sepharose CL-4B and Mono Q column chro - matographies with a recovery yield of 60% The molecular weight of the purified rHSA was estimated to be about 65,000 Da by denaturing SDS-PAGE The N terminal amino acid sequence of the purified HSA determined by Edman degradation was turned out to be Asp- Ala- His- Lys- Ser- Glu- Ala, suggesting that the rHSA expressed in H, polymorpha was efficiently secreted and correctly processed at the cleavage site of secretion signal sequence. The purified human albumin showed the pI value identical to that of authentic human serum albumin.

• The Korean Journal of Ecology
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• v.11 no.3
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• pp.145-152
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• 1988

The potential natural vegetation of Naejangsan national park area, southwestern Korea, was inferred from the actual vegetation. With the phytosociological classification, ordinatins and field surveys, the actual vegetation map of the area was made in scale 1:25, 000, including ten communities of Pinus densiflora, quercus mongolica, Quercus variabilis, Carpinus laxiflora, Daphnipyllum macropodum, Carpinus tschonoskii, Quercus aliena-Carpinus tschonoskii, Cornus controversa-Lindera erythrocarpa, Torreya mucifera-Zelkova serrate and Acer mono-Zelkova serrata community. The analyses of species richness, age structure and various informations on vegetation changes suggest the three pathways of late stage succession from P. densiflora forest to climatic climax. The first of them is through Q. variabilis forest to Q. monogolica forest in the upper parts of the mountain, the second through Q. variabilis and Q. serrata forest to C. laxiflora forest in the middle parts and the third through Q. aliena forest to C. tschonoskii forest in lower parts. Considering the actual vegetation and informations on the vegetation changes including human activities, the potential natural vegetation of the mountain mainly composed of Q. monogolica, C. laxiflora, C. tschonoskii, P. densiflora and Z. serrata forest as climatic climax and/or edaphic climax was inferred. The present situration of nature conservation in the area was estimated by the examination on the actual vegetation and potential natural vegetation map.

• Korean Journal Plant Pathology
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• v.10 no.3
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• pp.215-221
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• 1994

Polygalacturonase (PG) produced by Botrytis cinerea in the culture broth containing citrus pectin as a carbon source was partially purified and characterized. PG was produced on a range of carbon sources such as starch, glycerol, cellobiose, and Na+-PAG with total activities of 34.8, 32.0, 29.2, 27.8 units, respectively. The specific activity was highest with 2316.7 units on Na+-PGA. Proteins of culture filtrate were concentrated with polyethylene glycol and acetone and applied to a hydroxyapatite column. Among three active fractions collected from the column, the reaction containing the highest PG activity was resolved by a Q-sepharose column. The active fraction from the Q-sepharose column was further purified by HPLC Mono Q column. The partially purified enzyme was analyzed by 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Among a few protein bands revealed, the amount of the protein of which molecular weight estimated to be 43 kDa coincided with the PG activity. The partially purified PG had optimal temperatures between 35~55$^{\circ}C$ and pH between 4.5~5.5.

• Proceedings of the Korean Vacuum Society Conference
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• 2013.08a
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• pp.284-284
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• 2013

중 에너지 이온 산란 분석법(Medium Energy Ion Scattering Spectrometer, MEIS)은 50~500 keV로 이온을 가속 후 시료에 입사시켜 시료의 원자와 핵간 충돌로 산란되는 일차이온의 에너지를 측정하여 시료를 분석하는 기법으로, 원자층의 깊이 분해능으로 초박막의 표면 계면의 조성과 구조를 분석 할수 있는 유용한 미세 분석기술이다. 본 실험에서 에너지 70~100 keV의 He+ 이온을 사용하여 Pulse Width 1 ns의 Pulsed ion beam을 만들어 Start 신호로 사용하고 Delay-line-detector에 검출된 신호를 End 신호를 이용한 TOF-MEIS System을 개발하였다. 활용 가능한 분석시편으로 Ultra thin film 시편으로 1, 1.5, 2, 2.5, 3, 4 nm의 HfO2, 1.8, 4nm의 SiO2 시편을 분석 하였으며 Ultra Shallow Junction 시편으로 As Doped Si, Cs Doped Si 시편 및 Composition, Core/shell 구조의 Q-dot 시편으로 CdSe, CdSe/ZnS등 다양한 분석 실험을 진행 하였다. Composition, Core/shell 구조의 Q-dot 시편은 Diamond Like Carbon(DLC)의 Substrate에 Mono-layer로 형성하여 분석하였다.

• 한국생물공학회:학술대회논문집
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• 2001.11a
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• pp.221-224
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• 2001

Effect of hyperosmotic pressure on growth of recombinant Chinese hamster 。 vary cells and Erythropoietin (EPO) production was investigated. Cells were cultivated in batch modes at various osmolalities. When the osmolality increased from 314 to 463mOsm/Kg, specific EPO productivity (qp) was increased up to 1.6-fold but cell growth was inhibited. EPO has a complex oligosaccharide structure that plays an important role in biological activity in vivo. To investigate the influence of hypoerosmotic pressure on the glycosylation, structural analysis of oligosaccharide was calTied out. Recombinant human EPO was produced by CHO cells grown under various osmotic pressure and purified from culture supernatants by heparin-sepharose affinity column and immunoaffinity column. N-linked oligosaccharides were released enzymatically and isolated by paper chromatography. The isolated oligosaccharides were labeled with fluorescent dye, 2-aminobenzamide and analyzed with MonoQ anion exchange chromatography and GlycosepN amide chromatography for the assignment of GU (glucose unit) value. Glycan analysis by HPLC showed that neutral (asialo) oligosaccharide was increased slightly with an increase in osmolality. In portion of sialylated glycan, total relative amount of mono- and di-sialyated glycan was increased but that of tri- and tetra-sialylated glycan decreased as osmolality was increased.